This new history (below) leads
towards evidence of what the actual
"disease," Relapsing Fever
with, apparently
added,
OspA, really
is, immunologically and pathologically
(extra, activated, previously latent
viral infections).
We know
"Chronic Lyme" isn't just
IDSA's dozens of reports on "persisting spirochetes
past treatment."
We now
know these diseases sets to be
the non-inflammatory outcomes of
mycoplasmal-hijacked O2 and mycoplasmal-hijacked
sugars
from RBCs (erythrocytes), resulting in Chronic
Fatigue without anemia, plus, the
activation of Epstein-Barr. These
bad outcomes
of OspA-vaccination (or the autovaccination of spirochetal
blebbing) are IDSociety.org's "new Great Imitator"
outcomes.
The bad guys are correct to say "Lyme
disease" has only a temporary bad-knee
outcome, because "Lyme Disease" is like a
flea-circus.
It's a show - an
illusion - that the bad guys made up
themselves to go with OspA as a vaccine.
They did this - falsified the
diagnostics
for OspA-Borreliosis - to
set up later events, namely a monopoly
on all vector borne diseases (funding-
see Crazy
Eddie and his
girlfriend,
Durland Fish) as a "Vaccines and
Test Kits" "enterprise."
You can be assured of this because OspA
and B were left out of
the diagnostic
standard superimaginated at Dearborn (Steere
in Europe). You never test for the
outcome of a vaccine with the same
vaccine antigens used in the vaccine
Seronegative Epstein-Barr (4
articles), which is now known as Chronic
Fatigue/Fibromyalgia:
Down-regulation of MHC class II
expression through... [J Immunol. 2009]
- PubMed - NCBI
http://www.ncbi.nlm.nih.gov/pubmed/19201831
NEW
CHRONOLOGY OF THE DATA AVAILABLE THAT
DEMONSTRATES that
Pam3Cys-like INDUCED
IMMUNE SUPPRESSION and
the ACTIVATION OF EPSTEIN-BARR is
"CHRONIC LYME DISEASE."
1950s, Rockefeller Bioweapons
University (recall that the
Rockefellers are NAZIs and believe in
the same goal of extermination of
useless mouths to feed and especially
those with mental defects, even going so
far as to create them with marijuana and
childbood vaccines):
Fungal
antigens like LYMErix or OspA or
Borrelial outer surface proteins are
known to activate viruses via
immunosuppression:
"In Swiss mice, animals with high natural resistance to hepatitis virus, the pathogenicity of this agent was markedly enhanced by combined infection with eperythrozoa. Eperythrozoa were maintained throughout 18 successive passages in normal Princeton and Swiss weanlings with intact spleens. The combined infection of Princeton mice with eperythrozoa and the virus component of Gledhill, Dick, and Andrewes, which is nearly inactive when injected alone, resulted in acute hepatitis with fatal outcome."
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2136329/?tool=pubmed
How far back in time did
the likes of
Ray Dattwyler know about
fungal-antigen-induced immunosuppression?
This was one of Dattwyler's references:
1976;
Suppressor thymus-derived lymphocytes in
fungal infection.
Thymus-derived lymphocyte (T-cell) function, as determined in vivo by cutaneous reactivity to several antigens and in vitro by responsiveness to mitogens and antigens, was assessed in 14 patients infected with a variety of fungal organisms. While all patients manifested a normal frequency of peripheral blood T cells, only seven patients reacted to at least one of the antigens used for cutaneous testing and demonstrated normal in vitro T proliferative responses. Three patients exhibited cutaneous anergy but normal in vitro T-cell reactivity while four patients demonstrated persistent anergy and marked in vitro T-cell hyporeactivity which was independent of activity of infection, concurrent medication, or any associated disorders. The marked diminution of in vitro T-cell reactivity noted for these later four patients was not due to a deletion of antigen- or mitogen-reactive cells. Thus, patients' cells which had been initially cultured for 7 days without any mitogenic or antigenic stimulus and which were subsequently washed and recultured with phytohemagglutinin, concanavalin A, or histoplasmin demonstrated a marked increase in their responsiveness. Moreover, this reactivity noted for recultured cells could be suppressed by a nonphagocytic, nonadherent, nonimmunoglobulin-bearing, sheep red blood cell rosette-forming population of cells isolated from the fresh peripheral blood mononuclear cells of the same patient. While these "regulator" T cells were capable of suppressing T-proliferative responses to antigens and mitogens, they did not diminish pokeweed mitogen-induced immunoglobulin synthesis by normal bone marrow-derived lymphocytes. Patients in whom suppressor "T" cells were found were at risk for relapsing, disseminated fungal infection.
Look at the references here (dates, topics):
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC436656/pdf/jcinvest00145-0081.pdf
http://www.ncbi.nlm.nih.gov/pubmed/1082888
1981 (see
Dodd, Kennedy and Plum Island re
Newcastle Disease)
Suppression of the humoral immune response to inactivated Newcastle disease
virus by Mycoplasma meleagridis in the Turkey.
Groups of Mycoplasma meleagridis-free and -infected turkeys were vaccinated against Newcastle disease virus (NDV) at 5 and 8 weeks of age. The two groups did not differ significantly in antibody response to NDV as measured by the hemagglutination-inhibition test when a live vaccine (TCND) was used, but the two groups differed significantly when an inactivated vaccine was used; titers were higher in the mycoplasma-free group in both primary and secondary responses. The difference between responses of the mycoplasma-free and -infected groups to inactivated and live NDV was explained by a current hypothesis about the ontogeny of B-cell differentiation in the bursa of Fabricius and by its interference by M. meleagridis.
http://www.ncbi.nlm.nih.gov/pubmed/7337612
Ah, yes. The fabulous advantage of Brucellar/Mycoplasmal Pam3Cys-like antigens in antibody production... (See my "KMDickson" YouTube Channel for more on this from Don Scott)
1983,
PAM3CYS first synthesized
Synthesis of the mitogenic
S-[2,3-bis(palmitoyloxy)propyl]-N-palmitoylpentapeptide from Escherichia coli
lipoprotein.
The N-terminal pentapeptide of the
lipoprotein from the outer membrane of Escherichia coli was obtained by
coupling S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-N-palmitoyl-(R)-cysteine to
O-tert-butylseryl-O-tert-butyl-seryl-asparaginyl-alanine tert-butyl ester
followed by deprotection with trifluoroacetic acid. The tetrapeptide was
built up from alanine tert-butyl ester with N-9-fluorenylmethyloxycarbonyl
protected amino acids. S-[2,3-Bis(palmitoyloxy)propyl]-N-palmitoylcysteine
was obtained from N,N'-dipalmitoylcystine di-tert-butyl ester via reduction
to the thiol, and S-alkylation with racemic 3-bromo-1,2-propanediol followed
by esterification with palmitic acid in the presence of
dicyclohexylcarbodiimide/dimethylaminopyridine and deprotection with
trifluoroacetic acid. The compounds were characterized unequivocally by
13C-NMR and mass spectra. The diastereomers of
S-[2,3-bis(palmitoyloxy)propyl]-N-palmitoylcysteine tert-butyl ester with
opposite configuration at the propyl-C-2 atom could be separated on a
silica-gel column. http://www.ncbi.nlm.nih.gov/pubmed/6347861
1986-1994, The first diagnostic standard for Lyme Relapsing Fever designed
by Allen Steere:
Antigens of Borrelia burgdorferi recognized during Lyme disease.
Appearance of a new immunoglobulin M response and expansion of the
immunoglobulin G response late in the illness.
Using immunoblots, we identified
proteins of Borrelia burgdorferi bound by IgM and IgG antibodies during
Lyme disease. In 12 patients with early disease alone, both the IgM and
IgG responses were restricted primarily to a 41-kD antigen. This limited
response disappeared within several months. In contrast, among six
patients with prolonged illness, the IgM response to the 41-kD protein
sometimes persisted for months to years, and late in the illness during
arthritis, a new IgM response sometimes developed to a 34-kD component
of the organism. The IgG response in these patients appeared in a
characteristic sequential pattern over months to years to as many as 11
spirochetal antigens. The appearance of a new IgM response and the
expansion of the IgG response late in the illness, and the lack of such
responses in patients with early disease alone, suggest that B.
burgdorferi remains alive throughout the illness.
http://www.ncbi.nlm.nih.gov/pubmed/3531237
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC423723/?tool=pubmed
1990, CDC published standard is the one above, Steere's: Perform serial or sequential
Western Blots to "look for new IgM bands, because the appearance of new IgM
bands means the organism is still alive":
http://www.actionlyme.org/CDC_DOCUMENTS_1990.htm
1986, CDC officer Alan Barbour reveals
(transexual Lyme,
congenital Lyme in mice):
Biology of Borrelia
species.
http://www.ncbi.nlm.nih.gov/pubmed/3540570
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC373079/pdf/microrev00055-0033.pdf
1988,
Distinction between HIV-1 and HIV-2
infection using novel synthetic lipopeptide conjugates as antigens in enzyme
immunoassays.
“A
novel immunoassay technique using synthetic lipopeptide (Pam3Cys-Ser) linked to
immunodominant peptide domains of HIV-1 and HIV-2 envelope proteins as an
antigen adsorbent has been developed. Attachment of peptides to microtiter
plates can be considerably improved with this method by employing the
hydrophobic properties of lipopeptide. From the sera of 121 HIV-1 infected
patients 117 reacted with Pam3Cys-Ser-[HIV-1(598-609)cyclic disulfide]. Five of 5 HIV-2 positive sera were positive with
Pam3Cys-Ser-[HIV-2(593-603)cyclic disulfide]. Control sera failed to react
with these conjugates.”
http://www.ncbi.nlm.nih.gov/pubmed/2464607
People with HIV have antibodies to this, and as we will see later, apparently it was put into a vaccine for HIV.
1988,
Modulation of natural killer cell
activity by Borrelia burgdorferi.
FULL TEXT HERE
Golightly M,
Thomas J,
Volkman D,
Dattwyler R.
Department of Pathology, State University of New York, Stony Brook 11794.
PMID: 3056196 [PubMed
- indexed for MEDLINE]
Ann N Y Acad Sci. 1988;539:103-11.
"In summary, B. burgdorferi can induce a severe
inhibition both
in
vivo and in vitro in NK cell cytotoxic
capabilities. This is in contrast to
other bacteria that are known to activate NK cells. While the inhibition appears
to be directly related to spirochetal proliferation or concentration, the exact
mechanism is unclear and is
under active investigation. Interleukin-2
stimulation abrogates this inhibitory effect, possibly via the induction of LAK
cells, which may be involved secondarily to the inhibited endogenous NK. More
studies will be needed to elucidate these interactions and their full
significance to the host-microorganism interaction.
http://www.ncbi.nlm.nih.gov/pubmed/3056196"
I mentioned this NK-cell
immunosuppression to the FDA in Jan, 2001:
http://www.fda.gov/ohrms/dockets/ac/01/slides/3680s2_11.pdf
Report
above cited by 6 articles in MedLine:
http://www.ncbi.nlm.nih.gov/pubmed?db=pubmed&cmd=link&linkname=pubmed_pubmed_citedin&uid=3056196
FROM WIKIPEDIA:
Natural killer
Natural killer T cells (NKT cells) are a special kind of lymphocyte that bridges the adaptive immune system with the innate immune system. Unlike conventional T cells that recognize peptide antigen presented by major histocompatibility complex (MHC) molecules, NKT cells recognize glycolipid antigen presented by a molecule called CD1d. Once activated, these cells can perform functions ascribed to both Th and Tc cells (i.e., cytokine production and release of cytolytic/cell killing molecules). They are also able to recognize and eliminate some tumor cells and cells infected with herpes viruses.
1988, Raymond Dattwyler, et al,
"Modulation of natural killer cell activity by Borrelia burgdorferi."
"However, this does not explain the significantly decreased NK cytotoxic ability of these patients' PBL. It is possible that this inhibitor of NK activity is the result of a recruitment of inactive NK cells expressing CD16+. Leu-7- cells have cytotoxic activity (26). Alternatively, the decrease in NK activity may be due to a direct effect of the B. burgdorferi organism or its products on the NK cell. It has been reported that LPS isolated from Salmonella has the ability to inhibit the bacterial augmentation of activated killer cells while leaving endogenous NK. It is also possible that B. burgdorferu produces a toxin that is responsible for the inhibition. An adenylate cyclase toxin can be extracted from Bordetella that will increase cAMP levels when added to NK cells, resulting in an inhibition of NK activity without killing the NK effector cells.(27)"
http://www.ncbi.nlm.nih.gov/pubmed/3056196
Ie., Cells normally tasked to eradicate cells infected with herpes viruses have, according to Ray Dattwyler, et al, been somewhat chemically inhibited after exposure to, possibly, a Bb "toxin."
Note
that these two ▲▼ reports were
made at the same time - because they're
related- Seronegative Lyme and the
fungal-antigen influence
1988;
Seronegative Lyme disease.
Dissociation of specific T- and B-lymphocyte responses to Borrelia burgdorferi.
N Engl J Med.
1988 Dec 1;319(22):1441-6.
Dattwyler RJ,
Volkman DJ,
Luft BJ,
Halperin JJ,
Thomas J,
Golightly MG.
Department of Medicine, State University of New York, School of Medicine, Stony
Brook 11794-8161.
OspA
(Pam3Cys) -induced immunosuppression fits the model about which Dattwyler
complains, here.
The bad guys are furious (scared to death of homicide charges) because they know
they lied about the outcomes of LYMErix.
Neither
team could read their western Blots in OspA-vaccinated people.
Transcribed:
"This disorder in these seronegative
patients reflected a dissociation
between T-cell and B-cell immune
responses, in which the cell mediated arm
of the immune response was intact yet
the humoral portion of the response
appeared to be blunted. This
diminished antibody response is in
contrast to the T-cll anergy commonly
observed in several chronic infections
(e.g., infection with Mycobacteria
leprae or M. marinum, filariasis, and
some chronic fungal infections 29-33)."
http://www.ncbi.nlm.nih.gov/pubmed/3054554
1988: Lyme and
Multiple
Sclerosis and Roland Martin, the German recruit, used/abused to spin Lyme
and not find the MS-connection, which was Pam3Cys, after all:
*
Martin R on autoimmune T cells in the CSF of
Borreliosis victims (full text scanned in)
1988;
Enzyme-linked immunosorbent assay using recombinant OspC and the
internal 14-kDa flagellin fragment for serodiagnosis of early Lyme disease.
The
outer surface protein C (OspC) and the
internal 14-kDa flagellin fragment of
strain GeHo of Borrelia burgdorferi
sensu stricto were expressed as
recombinant proteins in Escherichia coli
and were purified for use in an
immunoglobulin M (IgM) enzyme-linked
immunosorbent assay (OspC-14-kDa antigen
ELISA). No hint at disturbing
protein-protein interferences, which
might influence the availability of
immunoreactive epitopes, was found when
the recombinant antigens were combined
in the ELISA. The recombinant
OspC-14-kDa antigen ELISA was compared
to a commercial IgM ELISA that used a
detergent cell extract from Borrelia
afzelii PKo as the antigen. According to
the manufacturer's information, the cell
extract contains, in addition to other
antigens, the following diagnostically
relevant antigens: the 100-kDa
(synonyms, 93- and 83-kDa antigens),
41-kDa, OspA, OspC, and 17-kDa antigens.
The specificity was adjusted to 95% on
the basis of data for 154 healthy
controls. On testing of 104 serum
samples from patients with erythema
migrans (EM), the sensitivity of the
recombinant ELISA (46%) for IgM
antibodies was similar to that of the
commercial ELISA (45%). However, when
42 serum samples from patients with
polyclonal B-cell stimulation due to an
Epstein-Barr virus infection were
tested, false-positive reactions were
significantly less frequent in the
recombinant ELISA (10%) than in the
whole-cell-extract ELISA (23%). OspC
displays sequence heterogeneity of up to
40% according to the genomospecies.
However, when the reactions of serum
specimens from controls and EM patients
with OspC from representative strains of
B. burgdorferi sensu stricto (strain
GeHo) and B. afzelii (strain PKo) were
compared in an ELISA, almost no
differences in specificity and
sensitivity were seen. This demonstrates
that the sera predominantly recognize
the common epitopes of OspC tested in
this study. In conclusion, we suggest
that the OspC-14-kDa antigens ELISA is a
suitable test for the detection of an
IgM response in early Lyme disease.
http://www.ncbi.nlm.nih.gov/pubmed/9542898
Full Text:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC104650/?tool=pubmed
Cited by:
http://www.ncbi.nlm.nih.gov/pubmed?db=pubmed&cmd=link&linkname=pubmed_pubmed_citedin&uid=9542898
RELATED:
Toll-like receptor agonists synergistically increase proliferation and activation of B cells by epstein-barr virus.
Mycoplasma synergize with HIV, too: PubMed: mycoplasma and HIV, link removed.
Lipid-associated membrane proteins of Mycoplasma fermentans and M. penetrans activate human immunodeficiency virus long-terminal repeats through Toll-like receptors.
Epstein-Barr virus induces MCP-1 secretion by human monocytes via TLR2.
It may be possible that when people with Borreliosis test positive to Epstein-Barr, it is because they actually have activated Epstein-Barr and that this is not a cross-reaction.
Search All (Borrelia and Epstein-Barr): [Borrelia and Epstein-Barr, PubMed searchremoved]
2010; Evidence of multiple infectious agents in mycosis fungoides lesions.
The etiology of mycosis fungoides (MF) remains to be determined. Several studies have proposed a viral etiology with controversial results. In this case-control study we investigated the presence of Epstein-Barr virus (EBV) and the debated presence of Human T-cell lymphotrophic virus I (HTLV-I) sequences, by polymerase chain reaction on nucleic acid extracts from formalin-fixed paraffin-embedded skin biopsies. Moreover, by a multivariate approach we analyzed in the same case-control study also the contribution of two previously examined pathogens: Hepatitis C virus (HCV) and Borrelia burgdorferi (Bb). Significant differences in the frequency of infectious agents in cases and controls were detected for Bb, HTLV-I and EBV. In MF patients we found the concurrent presence of two or three of these pathogen sequences in 21 out of 83 cases, but only in 1 out of 83 healthy controls. Our results suggest that the persistence of multiple infectious agents may cause a long-term antigenic stimulation contributing to the malignant transformation of T lymphocytes, especially when associated with HTLV-I like sequences. However, these infectious agents do not seem to have effects on disease progression.
http://www.ncbi.nlm.nih.gov/pubmed/20470773
1988;
Isolation and characterization of Borrelia burgdorferi-specific and
autoreactive T-cell lines from the cerebrospinal fluid of patients with Lyme
meningoradiculomyelitis.
http://www.ncbi.nlm.nih.gov/pubmed/2462820
Full Text
ONLY HERE
Later we will see that while there are
some patients who seem to have a true inflammatory response to Borrelial
antigens, others have MS-induced by Lyme as a result of the activated
Epstein-Barr, and in which case, they will have an aseptic meningitis,
one not detectable by antibody methods.
1988, Steere:
Anticardiolipin
antibodies in Lyme disease. (Note that Lupus is not a bad knee and that
Allen Steere owns this version of non-knee Lyme)
Sera from 28 patients with Lyme disease
were tested for the presence of anticardiolipin antibodies (ACLA). Seven
serum samples had elevated levels of IgM ACLA, and 4 had elevated levels of
IgG ACLA. Higher IgM ACLA positivity tended to be associated with neurologic
disease, and IgM ACLA levels correlated with the specific IgM response to
the infecting spirochete (P less than 0.01). Absorption experiments
indicated that ACLA and antispirochete antibodies are largely separate
populations. Thus, ACLA may occur in patients with Lyme disease,
particularly in those with neurologic abnormalities, and the production of
these antibodies seems to be linked to the specific IgM response.
http://www.ncbi.nlm.nih.gov/pubmed/3408508
FULL TEXT: LYME_AND_LUPUS_STEERE.htm
Later, Steere's
partner, Joe Craft, determined that Lupus-Lyme was likelier induced by
OspA/Epstein-Borreliosis:
1994: http://www.ncbi.nlm.nih.gov/pubmed/14707107
2010: http://www.ncbi.nlm.nih.gov/pubmed/20178121
"Although multiple mechanisms may be responsible for neurologic damage in Lyme disease, B. burgdorferi has been cultured from the cerebrospinal fluid of several patients with Lyme meningitis, suggesting a direct local infection (3). Furthermore, the occurrence of IgM ALCA seems to be linked to he specific IgM response, which occurs during the period of neurologic disease. Therefore, the apparent association with neurologic disease may be a co-linked variable without pathogenic implications." -- Allen Steere
Or, it could be a co-linked variable WITH pathogenic associations, such as activated Epstein-Barr. Activated by OspA-vaccination.
http://www.actionlyme.org/LYME_AND_LUPUS_STEERE.htm
1988;
Mitogenic activities of synthetic Escherichia coli lipid A and
a synthetic partial structure (tripalmitoyl pentapeptide) of E. coli
lipoprotein.
"Synthetic
Escherichia coli lipid A and synthetic S-[2,3-bis-(palmitoyloxy)propyl]-N-palmitoylpentapeptide
(tripalmitoyl pentapeptide [TPP]), representing the mitogenically active
principles of bacterial lipopolysaccharide (LPS) and lipoprotein, respectively,
were compared for their mitogenic activities on splenocytes of LPS responder (BALB/c)
and LPS-low-responder (C3H/HeJ) mice. Whereas lipid A was active only in
LPS-responder mice, TPP resulted in mitogenic activation of B lymphocytes from
both LPS-responder and LPS-low-responder mice. When the mitogens were added
simultaneously, mainly additive effects of both activators were observed. The
data suggest that two different B-lymphocyte populations are responding to these
two mitogens."
(Structure = Function; Function = Structure)
http://www.ncbi.nlm.nih.gov/pubmed/3281910
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC259838/?tool=pubmed
1988
[Primary lymphoma of the nervous
system following radiculoneuritis caused by Borrelia burgdorferi: study of a case]
A 50-year-old man had a primary lymphoma of
the central nervous system one year after a tick bite and a radiculoneuritis
secondary to Borrelia burgdorferi infection. Determination of antibodies
against Borrelia burgdorferi revealed increasing IgM and IgG titers, and the
autopsy showed a primary B-cell immunoblastic lymphoma of the brain without
evidence of extraneural lymphoproliferative disorder. Lymphoma of the brain
is a rare type of central nervous system cancer, and sporadic cases appear
without predisposing features such as immunosuppression or viral infection.
The controversy surrounding the histogenesis of this neoplasm is reviewed:
an unknown agent or a cofactor should provoke cellular proliferation and the
formation of a lymphoma. http://www.ncbi.nlm.nih.gov/pubmed/2451279
1989, Duray (in IDSA's journal),
Clinical pathologic
correlations of Lyme disease.
“Frank signs of meningeal
irritation herald stage II illness, reflected by an increase of CSF lymphocytes
and plasma cells and moderate increases in total protein in CSF [9,16,17].
Immature B cells can also be seen in the spinal fluid. These cells can
appear quite atypical- not unlike transformed or neoplastic lymphocytes.
Although it is known that spirochetes can be isolated from spinal fluid, they
are not recovered in all cases.”
http://www.ncbi.nlm.nih.gov/pubmed/2814170
IDSA_CLINIPATH_DURAY.htm ("Epstein-Barr-like transformed B
cells")
See
also Murine Gammaherpesvirus 68:
http://www.ncbi.nlm.nih.gov/pubmed?term=gammaherpesvirus[All%20Fields]%20AND%2068[All%20Fields]&cmd=DetailsSearch
Wondering whether a new/other/animal
herpesvirus could be causing these
"Epstein-Barr-like mutation" and illness
signs. Garth Nicolson also alludes to
this possibility.
1989, IDSA Reports The Incurable
Imitators.
1990,
Dave Dorward and Claude Garon at the NIH
reveal:
DNA Is Packaged within Membrane-Derived
Vesicles of Gram-Negative but Not
Gram-Positive Bacteria.
Recently, DNA packaged within nuclease-resistant membrane vesicles of Neisseria gonorrhoeae and Borrelia burgdorferi was described. This study assayed 18 species of gram-negative and gram-positive eubacteria for nuclease-protected DNA associated with extracellular membrane vesicles. Vesicles from only the gram-negative bacteria contained nuclease-protected linear or supercoiled DNAs or both.
http://www.ncbi.nlm.nih.gov/pubmed/16348232
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC184538/?tool=pubmed
This ▲ seems to be Allen Steere's version of Lyme disease [see also Dave Persing ("Target Imbalance")and Steere himself - "Chronic Lyme seems to be due to the persistence of the spirochete however few in number"] and more recently by Clifford Harding. Instead of it being true that there is an antigen-antibody complex (although they undoubtedly exist), it appears the antigen complexed to the HLA molecule is released as a free toxin.
1994; Dave Persing - Target imbalance: disparity of Borrelia burgdorferi genetic material in synovial fluid from Lyme arthritis patients.
Lyme arthritis is a late manifestation of Lyme disease that results in episodic synovial inflammation and swelling. Although this process is thought to be driven directly by the spirochetal etiologic agent, Borrelia burgdorferi, the organism itself has been recovered by culture only twice. In contrast, polymerase chain reaction (PCR) studies are usually positive. This apparent discrepancy in 19 culture-negative synovial fluid specimens from 18 patients with Lyme arthritis was investigated. In all 19, DNA sequences characteristic of plasmid-encoded genes OspA and OspB were easily detected. However, despite equivalent or even superior analytic sensitivity for detection of cultured organisms, the reactivity of two genomic DNA targets was often weak or absent altogether in the clinical specimens. This apparent overrepresentation of B. burgdorferi plasmid sequences was found exclusively in clinical specimens and not in cultured organisms. The physiologic imbalance of genomic and plasmid DNA reactivity in B. burgdorferi infection may signal an underlying pathogenetic mechanism. http://www.ncbi.nlm.nih.gov/pubmed
http://jid.oxfordjournals.org/content/169/3/668.long
2010, Dec, Clifford Harding: Mycobacterium tuberculosis synergizes with ATP to induce release of microvesicles and exosomes containing major histocompatibility complex class II molecules capable of antigen presentation.
Major histocompatibility complex class II (MHC-II) molecules are released by murine macrophages upon lipopolysaccharide (LPS) stimulation and ATP signaling through the P2X7 receptor. These studies show that infection of macrophages with Mycobacterium tuberculosis or M. bovis strain BCG enhances MHC-II release in synergy with ATP. Shed MHC-II was contained in two distinct organelles, exosomes and plasma membrane-derived microvesicles, which were both able to present exogenous antigenic peptide to T hybridoma cells. Furthermore, microvesicles from mycobacterium-infected macrophages were able to directly present M. tuberculosis antigen (Ag) 85B(241-256)-I-A(b) complexes that were generated by the processing of M. tuberculosis Ag 85B in infected cells to both M. tuberculosis-specific T hybridoma cells and naïve P25 M. tuberculosis T-cell receptor (TCR)-transgenic T cells. In the presence of prefixed macrophages, exosomes from mycobacterium-infected macrophages provided weak stimulation to M. tuberculosis-specific T hybridoma cells but not naïve P25 T cells. Thus, infection with M. tuberculosis primes macrophages for the increased release of exosomes and microvesicles bearing M. tuberculosis peptide-MHC-II complexes that may generate antimicrobial T-cell responses.
http://www.ncbi.nlm.nih.gov/pubmed/20837713
This reminds ▲ me of the Oct 4, 2003 Lyme RICO datapackage stolen of off AG/Senator Richard Blumenthal's desk by the crazy psycho DCF lying bitch Jessica Gauvin, because in it was an explainer about HLA-antigen kinetics and graphics from the murdered bioweapons scientists Don C. Wiley who predicted that H5N9 Swine Flu was likeliest to be the pandemic strain.... It is also seen in the Video on Biomarkers:
http://www.youtube.com/watch?v=6Pc8odvc9Z0
So, consider how useful that crazy psycho bitch Gauvin is to humanity.
2011; Update- More on Shed Blebs driving HLA-linked Dysimmunity:
Mycobacteria release active membrane vesicles that modulate immune responses in a TLR2-dependent manner in mice.
Bacteria naturally release membrane vesicles (MVs) under a variety of growth environments. Their production is associated with virulence due to their capacity to concentrate toxins and immunomodulatory molecules. In this report, we show that the 2 medically important species of mycobacteria, Mycobacterium tuberculosis and Mycobacterium bovis bacille Calmette-Guérin, release MVs when growing in both liquid culture and within murine phagocytic cells in vitro and in vivo. We documented MV production in a variety of virulent and nonvirulent mycobacterial species, indicating that release of MVs is a property conserved among mycobacterial species. Extensive proteomic analysis revealed that only MVs from the virulent strains contained TLR2 lipoprotein agonists. The interaction of MVs with macrophages isolated from mice stimulated the release of cytokines and chemokines in a TLR2-dependent fashion, and infusion of MVs into mouse lungs elicited a florid inflammatory response in WT but not TLR2-deficient mice. When MVs were administered to mice before M. tuberculosis pulmonary infection, an accelerated local inflammatory response with increased bacterial replication was seen in the lungs and spleens. Our results provide strong evidence that actively released mycobacterial vesicles are a delivery mechanism for immunologically active molecules that contribute to mycobacterial virulence. These findings may open up new horizons for understanding the pathogenesis of tuberculosis and developing vaccines.
http://www.ncbi.nlm.nih.gov/pubmed/21364279
1990, Kaiser-Permanente makes an
agreement with the DNA patenteers to say
"Lyme is nothing and needs no treatment, but is preventable with OspA
vaccination." The ALDF.com is founded by
mostly Israelis and Kaiser-Permanente in New York at NYMC, which is a Catholic
Medical School.
Kaiser remains to
this day at NYMC creating Kaiser-drones in the fashion of the US Military;
Kaiser will refund tuition to young MDs who promise to whore for Kaiser, and
lie as experts to the New York Medical Board. See the NY Medical
Board's lawyer, Ansel Marks reply to my
Conflict of Interest complaint - which included all the grants and
commercial products - in December 2000, which preceded the era of Pam
Weintraub's plagiarism.
Conflicts
Complaint, 1999 (not Weintraub):
http://groups.google.com/group/sci.med.diseases.lyme/msg/9afc48837708273c?hl=en&dmode=source
Said
Arthur Weinstein about the
cabal:
"The entrepreneurial trio are
Durland Fish, Ph.D., former director of
the College's Lyme Disease Center and
now a research scientist at Yale; Gary
P. Wormser, M.D., still professor of
medicine and pharmacology and chief of
the Division of Infectious Diseases at
the College; and John J. Connolly, Ed.D.,
former College president and current
chairman of the board of the American
Lyme Disease Foundation, Inc., which had
its genesis on the Valhalla campus in
1990."
1990, Pachner, Antigenic Variation in Brain-Invasive Spirochetes [FULL
TEXT]:
Borrelia burgdorferi infection of the brain: characterization of
the organism and response to antibiotics and immune sera in the mouse model.
To learn more about the neurologic
involvement in Lyme disease, we inoculated inbred mice with the causative
agent of Lyme disease, Borrelia burgdorferi. We cultured brains and other
organs, and measured anti-B burgdorferi antibody titers. We further studied
a brain isolate for its plasmid DNA content and its response in vitro to
immune sera and antibiotics. One strain of B burgdorferi, N40, was
consistently infective for mice, and resulted in chronic infection of the
bladder and spleen. SJL mice developed fewer culture-positive organs and had
lower antibody titers than Balb/c and C57Bl/6 mice. Organism was cultured
from the brain early in the course of infection, and this isolate, named
N40Br, was further studied in vitro. The plasmid content of N40Br was
different from that of the infecting strain, implying either a highly
selective process during infection or DNA rearrangement in the organism in
vivo. N40Br was very sensitive to antibiotics, but only after prolonged
incubation. Immune sera from both mice and humans infected with B
burgdorferi were unable to completely kill the organism by
complement-mediated cytotoxicity. These data demonstrate that B burgdorferi
infects the brain of experimental animals, and is resistant to immune sera
in vitro but sensitive to prolonged treatment with antibiotics.
http://www.ncbi.nlm.nih.gov/pubmed/2215944
See which Lyme crooks have referenced the
above article:
http://www.ncbi.nlm.nih.gov/pubmed?db=pubmed&cmd=link&linkname=pubmed_pubmed_citedin&uid=2215944
1990, May, Halperin, Dattwyler, et al;
Immunologic reactivity against
Borrelia burgdorferi in patients with motor neuron disease (ALS,
FULL TEXT)
Of 19 unselected patients with the
diagnosis of amyotrophic lateral sclerosis (ALS) living in Suffolk County,
New York (an area of high Lyme disease prevalence), 9 had serologic evidence
of exposure to Borrelia burgdorferi; 4 of 38 matched controls were
seropositive. Eight of 9 seropositive patients were male (8 of 12 male
patients vs 2 of 24 controls). Rates of seropositivity were lower among
patients with ALS from nonendemic areas. All patients had typical ALS; none
had typical Lyme disease. Cerebrospinal fluid was examined in 24 ALS
patients--3 (all with severe bulbar involvement) appeared to have
intrathecal synthesis of anti-B burgdorferi antibody. Following therapy with
antibiotics, 3 patients with predominantly lower motor neuron abnormalities
appeared to improve, 3 with severe bulbar dysfunction deteriorated rapidly,
and all others appeared unaffected. There appears to be a statistically
significant association between ALS and immunoreactivity to B burgdorferi,
at least among men living in hyperendemic areas.
http://www.ncbi.nlm.nih.gov/pubmed/2334308
These patients are "seronegative" or don't test positive to the Steere
Standard, which came later (1992), and 1994 (Dearborn):
None of these ▲ALS-Lyme cases would produce
a positive ELISA, which means they would be missed and go on to death from
ALS. Later we see the reason why Lyme produces the mycoplasmal
invasion known to be the cause of ALS (tolerance to and downregulation of
immune response to fungal antigens like mycoplasmal antigens and Borrelial
OspA).
This is the reason JJ Halperin tried to
pull an end-run around Blumenthal's subpoena (with
his "Neurology Guidelines") for all the crooks' own self-incriminating
data, which they refused to turn over, and which Blumenthal knew they had
produced.
It's a murder charge. They knew this
was the result of chronic, inadequately treated Lyme.
2002; ALS in Gulf War Illness Victims Due to Mycoplasma:
High frequency of systemic mycoplasmal infections in Gulf War veterans and civilians with Amyotrophic Lateral Sclerosis (ALS).
The presence of systemic mycoplasmal infections in the blood of Gulf War veterans (n=8) and civilians (n=28) with Amyotrophic Lateral Sclerosis (ALS) and age matched controls (n=70) was investigated by detecting mycoplasma gene sequences with forensic Polymerase Chain Reaction (PCR) and back hybridization with a radiolabeled internal oligonucleotide probe. Almost all ALS patients (30/36 or approximately 83%) showed evidence of Mycoplasma species in blood samples, whereas <9% of controls had blood mycoplasmal infections (P<0.001). Using PCR ALS patients with a positive test for any mycoplasmal infection were investigated for the presence of M. fermentans, M. pneumoniae, M. hominis and M. penetrans in their blood. All Gulf War veterans with ALS were positive for M. fermentans, except one that was positive for M. genitalium. In contrast, the 22/28 civilians with detectable mycoplasmal infections had M. fermentans (13/22, 59%) as well as other Mycoplasama species in their blood, and two of the civilian ALS patients had multiple mycoplasma species (M. fermentans plus M. hominis). Of the few control patients that were positive, only two patients (2/70, 2.8%) were positive for M. fermentans (P<0.001). The results support the suggestion that infectious agents may play a role in the pathogenesis and/or progression of ALS, or alternatively ALS patients are extremely susceptible to systemic mycoplasmal infections.
http://www.ncbi.nlm.nih.gov/pubmed/12383408
2005, ALS:
[Determination of systemic infections due to Mycoplasma in patients with clinically defined amyotrophic lateral sclerosis]
INTRODUCTION: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of unknown origin which has been linked to chronic infections due to intracellular microorganisms.
AIMS: The purpose of this study was to identify species of Mycoplasma in blood samples from patients with clinically defined ALS by means of the polymerase chain reaction (PCR) method in comparison to healthy control subjects.
PATIENTS AND METHODS: We conducted a case-control study involving 75 participants, 20 of whom were patients with clinically defined ALS and 55 healthy controls. Venous blood samples were taken and processed in the Neuroimmunology Laboratory, where they were submitted to the PCR test for Mycoplasma sp.RESULTS: The patients with ALS were between 35 and 82 years old (mean: 52.5); the ages of the healthy control subjects ranged from 35 to 60 years (mean: 44.1). After performing the PCR for Mycoplasma sp. the following results were obtained: among the patients with ALS, 10 were found to be positive (50%) and 10 were negative (50%), whereas in the control subjects we found six positives (10.91%) and 49 negatives (89.09%); these results were statistically significant (p = 0.001). On calculating the estimated risk, an odds ratio of 8167 (CI 95%: 2.4-27.6) was obtained. This indicates that the risk of suffering from ALS, if the PCR test for Mycoplasma sp. is positive, is 8:1.
CONCLUSIONS: There is a strong link between suffering from a chronic infection due to Mycoplasma and developing ALS. Intracellular pathogenic agents such as Mycoplasma can play a role in the genesis of neurodegenerative diseases.
http://www.ncbi.nlm.nih.gov/pubmed/16138281
1990, Radolf;
Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins.
The pathogenic spirochete
Borrelia burgdorferi contains a set of integral membrane proteins which were
selectively extracted into the detergent phase upon solubilization of intact
B. burgdorferi with the nonionic detergent Triton X-114. Virtually all of
these hydrophobic proteins were recognized by antibodies in pooled sera from
patients with chronic Lyme arthritis, demonstrating that proteins
partitioning into the detergent phase of Triton X-114 encompass the major B.
burgdorferi immunogens. Furthermore, most of these immunogenic proteins,
including the previously characterized OspA and OspB membrane antigens,
could be biosynthetically labeled when B. burgdorferi was incubated in vitro
with [3H]palmitate. The OspA and OspB antigens were
radioimmunoprecipitated from [3H]palmitate-labeled detergent-phase proteins
with monoclonal antibodies, and [3H]palmitate was recovered unaltered from
these proteins after sequential alkaline and acid hydrolyses. The
combined results provide formal confirmation that the major B. burgdorferi
immunogens extracted by Triton X-114 are lipoproteins. The demonstration
that B. burgdorferi integral membrane antigens are
lipoproteins may explain the basis of their immunogenicity and may help to
improve our understanding of the surface topology of B. burgdorferi. http://www.ncbi.nlm.nih.gov/pubmed/2318538
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC258571/pdf/iai00052-0147.pdf
1991, Rheumatology News,
Steere and “Seronegative Lyme”:
“I am convinced this entity exists and is
associated with Multiple Sclerosis."
“If B. burgdorferi does cause this
syndrome, it's absolutely amazing
that this spirochete would mimic not only rheumatoid arthritis (RA)
but also multiple sclerosis (MS), two of the most puzzling and
devastating autoimmune diseases.
“Now I would like to proceed to the issue of
seronegative
Lyme
disease. I am convinced this entity exists..." -- Allen Steere
1991 re SERONEGATIVE, EBV-ACTIVATING Lyme - and LYMErix/ OspA and IL-10 -, cited over 100 times
Interleukin 10 (IL-10) and viral IL-10 strongly reduce antigen-specific human T cell proliferation by diminishing the antigen-presenting capacity of monocytes via downregulation of class II major histocompatibility complex expression.
SourceDNAX Research Institute, Human Immunology, Palo Alto, California 94304.
AbstractInterleukin 10 (IL-10) and viral IL-10 (v-IL-10) strongly reduced antigen-specific proliferation of human T cells and CD4+ T cell clones when monocytes were used as antigen-presenting cells. In contrast, IL-10 and v-IL-10 did not affect the proliferative responses to antigens presented by autologous Epstein-Barr virus-lymphoblastoid cell line (EBV-LCL). Inhibition of antigen-specific T cell responses was associated with downregulation of constitutive, as well as interferon gamma- or IL-4-induced, class II MHC expression on monocytes by IL-10 and v-IL-10, resulting in the reduction in antigen-presenting capacity of these cells. In contrast, IL-10 and v-IL-10 had no effect on class II major histocompatibility complex (MHC) expression on EBV-LCL. The reduced antigen-presenting capacity of monocytes correlated with a decreased capacity to mobilize intracellular Ca2+ in the responder T cell clones. The diminished antigen-presenting capacities of monocytes were not due to inhibitory effects of IL-10 and v-IL-10 on antigen processing, since the proliferative T cell responses to antigenic peptides, which did not require processing, were equally well inhibited. Furthermore, the inhibitory effects of IL-10 and v-IL-10 on antigen-specific proliferative T cell responses could not be neutralized by exogenous IL-2 or IL-4. Although IL-10 and v-IL-10 suppressed IL-1 alpha, IL-1 beta, tumor necrosis factor alpha (TNF-alpha), and IL-6 production by monocytes, it was excluded that these cytokines played a role in antigen-specific T cell proliferation, since normal antigen-specific responses were observed in the presence of neutralizing anti-IL-1, -IL-6, and -TNF-alpha mAbs. Furthermore, addition of saturating concentrations of IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha to the cultures had no effect on the reduced proliferative T cell responses in the presence of IL-10, or v-IL-10. Collectively, our data indicate that IL-10 and v-IL-10 can completely prevent antigen-specific T cell proliferation by inhibition of the antigen-presenting capacity of monocytes through downregulation of class II MHC antigens on monocytes.
1992
1992, Paul Duray re-wrote/re-presented for IDSA’s Cold
Spring Harbor Conference:
"On occasion, these
atypical-appearing large lymphocytes have been misinterpreted in biopsy by
several laboratories as cells of a malignant lymphoma or leukemia. Bb
antigens, then, may stimulate growth of immature lymphocytic suibsets in
some target organs, as well as in the cerebrospinal fluid (Szyfelbein and
Ross 1988). Usual bacterial infections do not produce such lymphocytic
infiltrates in tissue. These immunoblastoid cells in Bb infections at
times resemble those found in Epstein-Barr virus infections. Does Bb
reactivate latent virus infections in tissues? Do some tick inocula
harbor simultaneous infectious agents (ixodid ticks can harbor Rickettsiae,
Babesia microti, and Ehrlichia bacteria, in addition to Bb), producing
multi-agent infections in some hosts? Further studies can clarify these
issues by mans of tissue-based molecular probe analysis." -
Paul
Duray, NCI, NIH, Ft. Detrick, at the 1992
Cold Spring Harbor Conference, published in Steve Schutzer's Lyme
Disease: Molecular and Immunologic Approaches.
http://www.amazon.com/Lyme-Disease-Immunologic-Approaches-Communications/dp/0879693770/ref=sr_1_2?ie=UTF8&s=books&qid=1214848669&sr=1-2
1992; Klempner and "Borrelia are un-eradicable":
Fibroblasts protect the Lyme disease spirochete, Borrelia burgdorferi, from ceftriaxone in vitro.
The Lyme disease spirochete, Borrelia burgdorferi, can be recovered long after initial infection, even from antibiotic-treated patients, indicating that it resists eradication by host defense mechanisms and antibiotics. Since B. burgdorferi first infects skin, the possible protective effect of skin fibroblasts from an antibiotic commonly used to treat Lyme disease, ceftriaxone, was examined. Human foreskin fibroblasts protected B. burgdorferi from the lethal action of a 2-day exposure to ceftriaxone at 1 microgram/mL, 10-20 x MBC. In the absence of fibroblasts, organisms did not survive. Spirochetes were not protected from ceftriaxone by glutaraldehyde-fixed fibroblasts or fibroblast lysate, suggesting that a living cell was required. The ability of the organism to survive in the presence of fibroblasts was not related to its infectivity. Fibroblasts protected B. burgdorferi for at least 14 days of exposure to ceftriaxone. Mouse keratinocytes, HEp-2 cells, and Vero cells but not Caco-2 cells showed the same protective effect. Thus, several eukaryotic cell types provide the Lyme disease spirochete with a protective environment contributing to its long-term survival.
http://www.actionlyme.org/MarkKlempner_Fibroblasts.htm (full text)
http://www.ncbi.nlm.nih.gov/pubmed/1634816
1990; Steere using this seronegative Lyme Assay to evaluate "Chronic Neurologic
Lyme" cases:
http://www.nejm.org/doi/pdf/10.1056/NEJM199011223232102
1991, Oct, Steere using the Seronegative Lyme Assay to determine his lab workers had been exposed:
The T-cell proliferative assay in the diagnosis of Lyme disease.
OBJECTIVE: To determine the sensitivity and specificity of the T-cell proliferative assay as a diagnostic test in Lyme disease.
DESIGN: Cross-sectional study of patients with Lyme arthritis or chronic neuroborreliosis who had a history of erythema migrans, positive antibody responses to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), or both; patients with other diseases; and healthy subjects.
SETTING: Diagnostic Lyme disease clinic in a university hospital.
PATIENTS: Forty-two of the 67 patients with active Lyme arthritis or chronic neuroborreliosis who were seen during the study period; 16 patients with inactive late Lyme disease; 77 patients with other rheumatologic or neurologic diseases; 9 workers from the Borrelia laboratory; and 9 healthy subjects.
MEASUREMENTS AND MAIN RESULTS: Nineteen of 42 patients with Lyme arthritis or chronic neuroborreliosis and 4 of 77 patients with other diseases had positive T-cell proliferative responses to B. burgdorferi antigens. The sensitivity of the proliferative assay was 45% (95% Cl, 30% to 60%) and the specificity was 95% (95% Cl, 87% to 99%). Twelve of 27 patients with active Lyme arthritis, 7 of 15 patients with chronic neuroborreliosis, 4 of 16 patients with inactive Lyme disease, 4 of 9 healthy Borrelia laboratory workers, and 0 of 9 healthy subjects had positive responses. Three of five patients with Lyme disease who had negative or indeterminant antibody responses by ELISA had positive T-cell proliferative responses.
CONCLUSION: The T-cell proliferative assay may be a helpful diagnostic test in the small subset of patients with late Lyme disease who have negative or indeterminant antibody responses by ELISA. http://www.ncbi.nlm.nih.gov/pubmed/1883122
Full Text of Dattwyler/Volkman Seronegative Lyme Assay:
http://www.actionlyme.org/STEERES_SERONEG_LYME_ASSAY.htm
1992; Autopsy report of primary CNS
B-cell lymphoma indistinguishable from multiple sclerosis: diagnosis with the
immunoglobulin gene rearrangements analysis.
We report a case of primary CNS B-cell
lymphoma indistinguishable from multiple sclerosis (MS). MRI of the head
showed the spontaneous disappearance of the white matter lesions and the
progressive cerebral atrophy. The brain biopsy failed to make a diagnosis of
CNS lymphoma but rather suggested MS. Although the primary CNS lymphoma was
suspected at autopsy, the immunohistochemical study showed the
CNS-infiltrating lymphoid cells comprising both T-cells and B-cells.
Analysis of the immunoglobulin and T-cell receptor gene rearrangements first
provided evidence of primary CNS B-cell lymphoma. http://www.ncbi.nlm.nih.gov/pubmed/1431983
1996, Monoclonal B-cell population mimicking lymphoma in a patient
with multiple sclerosis.
BACKGROUND: Diagnosis of
intraparenchymal brain lesions has usually required invasive diagnostic
procedures, because too few cells are shed into cerebrospinal fluid to
permit cytologic diagnosis. Polymerase chain reaction technology makes it possible to identify cell populations that are present at a much lower frequency than traditional techniques.
CASE REPORT: A young woman
presented with multiple brain lesions raised the question of primary
central nervous system lymphoma. Polymerase chain reaction analysis of
cerebrospinal fluid showed evidence of a monoclonal B-cell population
heightening suspicion of lymphoma. Brain biopsy showed acute
demyelination most consistent with multiple sclerosis.
CONCLUSION: Although T-cell
restriction has been demonstrated in multiple sclerosis lesions, the
finding of a monoclonal B-cell population was unexpected and to our
knowledge has not been previously reported. This case emphasizes that
monoclonality is not always indicative of a neoplastic process,
particularly in the central nervous system.
http://www.ncbi.nlm.nih.gov/pubmed/8629904
1992; Steere also gave a presentation at the Academy of Insurance
Medicine, which, of course, no one knew even existed, and he reversed
his previous position (1986) where the presence of IgM bands meant the
infection was ongoing:
◄Steere's
presentation at the Academy of Insurance Medicine in 1992
Compare to:
1990; Steere using this seronegative Lyme Assay to evaluate "Chronic Neurologic
Lyme" cases:
http://www.nejm.org/doi/pdf/10.1056/NEJM199011223232102
In 1986, before the RICO entity was established, Steere said to look for new IgM:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=423723&blobtype=pdf
1992; [The effect of Eperythrozoon suis infection on the
osmotic fragility of erythrocytes]
Osmotic fragility of erythrocytes was
tested in weaned pigs experimentally infected with Eperythrozoon (E.) suis.
Acute eperythrozoonosis of splenectomized pigs led to an increase of osmotic
fragility. It is supposed that E. suis infection causes a structural change
in erythrocyte membrane. Possible mechanisms of this cell membrane injury
are discussed.
http://www.ncbi.nlm.nih.gov/pubmed/1471973
1992: Picken
Phylogeny (Lyme is Relapsing Fever)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC265004/pdf/jcm00025-0121.pdf
1992: "A Bogus Article" regarding how OspA vaccination of mammals turns
them into walking canisters of tick-disinfectant. By this time, Fikrig and
Flavell already knew how to determine if there were spirochetes in ticks via the
Borrelia-specific Flagellin DNA method.
They did not have to use this bogus immunofluorescence of OspA ridiculous test:
Elimination of Borrelia burgdorferi from vector ticks
feeding on OspA-immunized mice.
Although recombinant outer surface protein
A (OspA) of Borrelia burgdorferi protects mice against injected Lyme disease
spirochetes, the mode of protection has not yet been explored. Indeed, the
efficacy of vaccine-induced immunity against a realistic vector-mediated
challenge remains unexplored. Accordingly, we determined whether this
immunogen protects mice against spirochetes delivered by nymphal Ixodes
dammini ticks. Following challenge by tick bite, no spirochetes could be
cultured from immunized mice, and no characteristic histopathology was
found. The spirochete was not detected in ticks that fed on immunized
animals and was present in virtually all ticks that fed on nonimmunized
mice. We conclude that OspA-immunized mice are protected from spirochetal
infection, at least in part, because the spirochete is destroyed in the
infecting tick.
http://www.ncbi.nlm.nih.gov/pubmed/1608951
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC49303/pdf/pnas01086-0226.pdf
1992, May; Macromolecular assemblage in the design
of a synthetic AIDS vaccine.
Defoort JP,
Nardelli B,
Huang W,
Ho DD,
Tam JP.
Rockefeller University, New York, NY 10021.
We describe a peptide vaccine model based on the mimicry of surface coat protein of a pathogen. This model used a macromolecular assemblage approach to amplify peptide antigens in liposomes or micelles. The key components of the model consisted of an oligomeric lysine scaffolding to amplify peptide antigens covalently 4-fold and a lipophilic membrane-anchoring group to further amplify noncovalently the antigens many-fold in liposomal or micellar form. A peptide antigen derived from the third variable domain of glycoprotein gp120 of human immunodeficiency virus type 1 (HIV-1), consisting of neutralizing, T-helper, and T-cytotoxic epitopes, was used in a macromolecular assemblage model (HIV-1 linear peptide amino acid sequence 308-331 in a tetravalent multiple antigen peptide system linked to tripalmitoyl-S-glycerylcysteine). The latter complex, in liposome or micelle, was used to immunize mice and guinea pigs without any adjuvant and found to induce gp120-specific antibodies that neutralize virus infectivity in vitro, elicit cytokine production, and prime CD8+ cytotoxic T lymphocytes in vivo. Our results show that the macromolecular assemblage approach bears immunological mimicry of the gp120 of HIV virus and may lead to useful vaccines against HIV infection.
http://www.ncbi.nlm.nih.gov/pubmed?term=1349173[uid]&cmd=DetailsSearch
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC525594/pdf/pnas01083-0219.pdf
1992, Sep;
A rational design of synthetic
peptide vaccine with a built-in
adjuvant. A modular approach for
unambiguity
We describe a peptide vaccine model containing a built-in adjuvant. This model used a multiple antigen peptide system (MAPS) to amplify peptide antigens and a lipoamino acid, tripalmitoyl glyceryl cysteine (P3C), as a built-in adjuvant. An 18-residue peptide antigen (B2) derived from the third variable domain (amino acid 312-329) of the glycoprotein gp120 of type I human immunodeficiency virus (HIV-1) was used in this model. This peptide antigen is a suitable target since it consists of neutralizing, T-helper, and T-cytotoxic epitopes. The peptide antigen in a tetravalent MAPS format (B2M-P3C) with a lipophilic attachment was synthesized by two routes for comparison: a direct stepwise approach and an indirect modular approach. In the stepwise approach, each residue was sequentially added to the peptide resin to give B2M-P3C and the P3C was incorporated to the side chain of a carboxyl terminal lysine as Fmoc-Lys(P3C). In the modular approach, a module containing a chloroacetylated core matrix of MAPS (M-P3C) with a carboxyl tetrapeptide bearing Lys(P3C) and a second module containing the peptide antigen B2 with a cysteine at its terminus were synthesized and purified separately, and then coupled to each other to form B2M-P3C. In the modular approach, the molecular ion of B2M-P3C was unambiguously identified by ion-spray mass spectrometry. B2M-P3C, administered in liposomes without any adjuvant such as Freund's complete adjuvant, was used to immunize mice and found to induce gp120-specific antibodies in vitro, and prime cytotoxic T lymphocytes in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)
http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=1478779[uid
1994;
Tolerance to lipopolysaccharide involves mobilization of nuclear factor
kappa B with predominance of p50 homodimers.
Stimulation of the human
monocytic cell line Mono Mac 6 with lipopolysaccharide (LPS) leads to rapid and
transient expression of cytokines like tumor necrosis factor (TNF). When such
cells are precultured for 2 days with a low dose of LPS (20 ng/ml) followed by
stimulation with a high dose of LPS (1 microgram/ml), expression of the TNF gene
is minimal, i.e. the cells are tolerant. In nuclear run-on analysis, such
tolerant cells show only a low degree of transcription, indicating that
tolerance operates at or upstream of the transcription level. The CD14 LPS
receptor is, however, up-regulated (not down-regulated) in tolerant cells, and
LPS can, in fact, still lead to activation of tolerant cells as evidenced by
mobilization of the transcription factor nuclear factor kappa B (NF-kappa B).
Resolution of the NF-kappa B complex in gel shift analysis shows that the
binding protein, mobilized in naive Mono Mac 6 cells, consists mainly of p50-p65
heterodimers, while in tolerant cells, the p50 homodimer is predominant. This
increase in p50 homodimers coincides with an increase in p105 mRNA, suggestive
of a transcriptional up-regulation of p50. Reporter gene analysis reveals that
the NF-kappa B complex mobilized in tolerant cells is functionally inactive in
that NF-kappa B-dependent luciferase constructs containing the human
immunodeficiency virus long terminal repeat or the TNF 5'-region show only
minimal transactivation after LPS stimulation. Similar to Mono Mac 6 cells,
primary blood monocytes, when precultured with a low dose of LPS, also become
tolerant and produce little TNF after LPS stimulation. The tolerant blood
monocytes also up-regulate CD14, and they mobilize NF-kappa B with a
predominance of p50 homodimers. Taken together, these results demonstrate that
tolerance to LPS is determined by post-receptor mechanisms that involve an
altered composition of the NF-kappa B complex.
http://www.ncbi.nlm.nih.gov/pubmed/7516328
http://www.jbc.org/content/269/25/17001.long
1995,
CDC Officer Alan Barbour patents Borrelia Theileri, which is aBovine Borreliosis or Cow Relapsing
Fever, but never tells Edwin Masters he patented Masters’ Disease right from
under Masters’ nose:
http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&l=50&s1=5,932,220.PN.&OS=PN/5,932,220&RS=PN/5,932,220
2001; Lone star tick-infecting borreliae are most closely related to the agent
of bovine borreliosis.
Although Borrelia theileri,
the agent of bovine borreliosis, was described at the turn of the century
(in 1903), its relationship with borreliae causing Lyme disease or relapsing
fever remains undescribed. We tested the previously published hypothesis
that spirochetes infecting Lone Star ticks (Amblyomma americanum) may
comprise B. theileri by analyzing the 16S ribosomal DNAs (rDNAs) and
flagellin genes of these spirochetes. B. theileri, the Amblyomma agent, and
B. miyamotoi formed a natural group or clade distinct from but most closely
related to that of the relapsing fever spirochetes. B. theileri and the
Amblyomma agent were 97 and 98% similar at the nucleotide level within the
analyzed portions of the 16S rDNA and the flagellin gene respectively,
suggesting a recent divergence. The agent of bovine borreliosis might be
explored as a surrogate antigen for the as-yet-uncultivatable Amblyomma
agent in studies designed to explore the etiology of a Lyme disease-like
infection associated with Lone Star ticks.
http://www.ncbi.nlm.nih.gov/pubmed/11158095
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC87764/?tool=pubmed
1995;
Interleukin-10 is upregulated in LPS tolerance.
Lipopolysaccharide (LPS) stimulation of the human monocytic cell line Mono
Mac 6 leads to rapid expression of both the pro-inflammatory cytokine tumor
necrosis factor (TNF) and the anti-inflammatory cytokine interleukin-10
(IL-10). Preculture of these cells with a low dose of LPS for 2 days
rendered the cells tolerant to subsequent stimulation, in that TNF gene
expression is only minimal, both at the mRNA and at the protein level. IL-10
shows a reciprocal pattern, however, as expression of this gene is
upregulated in precultured cells, and it will further increase upon
subsequent stimulation. Although TNF has been shown to induce IL-10, and
IL-10 was found to downregulate TNF, this reciprocal regulation does not
explain the pattern observed in LPS tolerance in Mono Mac 6, since
neutralizing antibodies against TNF and IL-10 could not prevent upregulation
of IL-10 and downregulation of TNF, respectively. Treatment of Mono Mac 6
cells during LPS preculture with interferon-gamma (IFN-gamma) could,
however, reverse tolerance: LPS/IFN-gamma precultured cells produced high
levels of TNF transcripts upon subsequent stimulation, while the response of
the IL-10 gene was attenuated. The data show that LPS tolerance does not
involve a passive downregulation of all types of monocyte functions, but it
is an orchestrated response with downregulation of pro- and upregulation of
anti-inflammatory cytokines.
http://www.ncbi.nlm.nih.gov/pubmed/7583353
1995, April,
(Radolf) Dermal inflammation elicited by
synthetic analogs of Treponema pallidum
and Borrelia burgdorferi lipoproteins.
The membrane lipoproteins of Treponema pallidum and Borrelia burgdorferi have potent immunostimulatory properties in vitro, implicating them as major inflammatory mediators in syphilis and Lyme disease. Recently, we reported that synthetic lipohexapeptide analogs (lipopeptides) of the lipoproteins could be used as surrogates for native spirochetal lipoproteins in immune cell activation studies in vitro. The present study was designed to evaluate the inflammatory properties of the lipopeptides in vivo and to correlate the cellular responses to these synthetic analogs with the histopathology of syphilis and Lyme disease. Lipopeptides corresponding to the 47-kDa major membrane lipoprotein of T. pallidum and the outer surface protein A of B. burgdorferi injected intradermally induced dose-dependent dermal inflammation in mice; the initial predominantly neutrophilic (mice) or heterophilic (rabbits) cellular infiltrates were followed by infiltrates consisting predominantly of mononuclear cells. The intradermal response of rabbits to the 47-kDa lipopeptide was strikingly similar to that observed for animals infected intradermally with T. pallidum. In all cases, lipopolysaccharide was substantially more potent as an inflammatory mediator than the spirochetal lipopeptides. In contrast to the lipopeptides, nonacylated hexapeptides elicited minimal or no dermal lesions in mice or rabbits, underscoring the importance of acyl modification to the inflammatory properties of the lipopeptides. This study provides the first in vivo evidence that the spirochetal lipoproteins/lipopeptides contribute to the immunopathogenesis of syphilis and Lyme disease.
A structural feature common to T. pallidum and B. burgdorferi is that the majority of their integral membrane proteins are lipid modified (9, 12, 42). Compelling data have now emerged supporting that these spirochetal lipoproteins are potent immunopotentiators. Treponemal and borrelial lipoproteins have been shown to activate monocytes/macrophages, B cells, and endothelial cells in vitro (1, 28, 29, 36, 39, 49, 56), suggesting that these molecules are inflammatory mediators in both syphilis and Lyme disease. More recently, we reported that synthetic lipohexapeptide analogs (lipopeptides) corresponding to the N termini of the native spirochetal lipoproteins could be used as lipoprotein surrogates in immune cell activation studies (16, 37). T
hese lipopeptides, modeled after earlier studies of Bessler, Jung, and coworkers on the murein (Braun’s) lipoprotein of Escherichia coli (21, 22), have been configured as N-palmitoyl-S-dipalmitoylglycerylcysteine-pentapeptides (16, 37).
http://www.ncbi.nlm.nih.gov/pubmed/7890417
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC173182/pdf/631507.pdf
1995;
Bacterial lipoproteins may substitute for cytokines in the humoral
immune response to T cell-independent type II antigens.
Bacterial lipoproteins share a common
structural motif that has been shown to stimulate proliferation and Ig
secretion of murine B cells, in a manner distinct from that mediated by LPSs.
Studies of lipoprotein-mediated B cell activation utilized heterogeneous
populations of lymphoid cells, leaving unresolved their ability to directly
activate resting B cells, as well as their ability to interact with other B
cell stimuli. Using highly enriched and/or sort-purified resting murine B
cells, we demonstrate that, in contrast to previous reports, lipoproteins
(lipoprotein-D, lipoprotein-OspA, and/or the synthetic analogue Pam3Cys)
stimulate little, if any, proliferation or Ig secretion in resting B cells.
However, when combined with a multivalent membrane (m)Ig-mediated
cross-linking signal, dextran-conjugated anti-IgD Abs (alpha delta-dex),
lipoproteins mediate up to 10,000-fold inductions in IgM secretion and up to
25-fold enhancements in cellular proliferation relative to that observed
with alpha delta-dex alone, in the absence of added cytokines. This mIg-mediated
enhancement of Ig secretion was not observed when B cells were stimulated
with bivalent, unconjugated anti-Ig. CD40 ligand (CD40L), shows a similar,
although somewhat more moderate, synergy with lipoproteins for induction of
proliferation and IgM secretion. By contrast, lipoproteins by themselves are
relatively ineffective at costimulating Ig secretion in the presence of
various combinations of cytokines. These data suggest that bacteria may
induce Ag-specific humoral immunity through the action of bacterial
polysaccharides that mediate an Ag-specific multivalent mIg signal, in
concert with bacterial lipoproteins that deliver ancillary signals, without
a requirement for recruitment of non-B cell types.
http://www.ncbi.nlm.nih.gov/pubmed/7499841
1995, Tully.
Spiroplasma ixodetis sp. nov., a new
species from Ixodes pacificus ticks collected in Oregon.
Eight strains of mollicutes were isolated from
pooled suspensions prepared from western black-legged ticks (Ixodes pacificus)
collected in Oregon. Morphologic examination by electron and dark-field
microscopic techniques showed that each strain consisted of a mixture of motile,
tightly coiled helical cells, small coccoid cells with diameters ranging from
300 to 500 nm, and pleomorphic, straight or branched filamentous forms. All
cellular forms were surrounded by a single cytoplasmic membrane, and there was
no evidence of a cell wall. The organisms were filterable and fastidious in
their growth requirements. The optimum temperature for growth was 30 degrees C,
but multiplication occurred at temperatures ranging from 23 to 32 degrees C. The
strains catabolized glucose but did not hydrolyze arginine or urea. The genome
size of strain Y32T (T = type strain) was 2,220 kbp, and the DNA base
composition (guanine-plus-cytosine content) of this organism was 25 +/- 1 mol%.
The eight isolates were serologically related to each other but were not related
to 37 other type or representative strains belonging to the genus Spiroplasma.
Strain Y32 (= ATCC 33835) is the type strain of Spiroplasma ixodetis sp. nov.
http://www.ncbi.nlm.nih.gov/pubmed/7857803
1995; Robert Schoen and Dave Persing report that Western Blots were
unreadable in OspA-vaccinated people:
Borrelia burgdorferi enzyme-linked immunosorbent assay for
discrimination of OspA vaccination from spirochete infection.
Recombinant Lyme disease vaccines based on
purified preparations of outer surface protein A (OspA) have been shown to
be effective in preventing transmission of Borrelia burgdorferi in
experimental animal models and are now being tested in humans. Since the
most widely used screening tests for Lyme disease are based on a whole-cell
sonicate of B. burgdorferi, serologic false positivity in vaccinated persons
could result from reactivity to OspA within the antigen preparation. In
order to avoid serologic false positivity in vaccinated subjects, we
developed an immunoassay based on a low-passage-number, naturally occurring
variant of B. burgdorferi which lacks the plasmid encoding OspA and OspB.
The use of an antigen preparation derived from this organism provided
sensitive and specific detection of B. burgdorferi seropositivity in
experimental animals and in human Lyme disease cases. The OspA-B-negative
enzyme-linked immunosorbent assay (ELISA) also appeared to be capable of
discriminating the vaccinated state from vaccine failure and natural
infection in experimental animals. Sera from human subjects participating in
a vaccine trial gave false-positive results with an ELISA based on an
OspA-containing strain, but no such reactivity was observed when the
OspA-negative ELISA was used. We conclude that low-passage-number
OspA-B-negative isolates in immunoassays may become useful for the
immunologic discrimination of the vaccinated state, natural infection, and
vaccine failure.
http://www.ncbi.nlm.nih.gov/pubmed/8968914
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC229545/pdf/350233.pdf
Method for detecting B. burgdorferi infection
http://patft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=/netahtml/PTO/srchnum.htm&r=1&f=G&l=50&s1=6045804.PN.&OS=PN/6045804&RS=PN/6045804
The method provided by the invention is
particularly useful to discriminate B. burgdorferi infection from OspA
vaccination, although it is sufficiently sensitive and specific to use in
any general Lyme disease screening or diagnostic application. Thus, the
method of the invention is particularly appropriate for large scale
screening or diagnostic applications where only part of the subject
population has been vaccinated or where the vaccination status of the
population is unknown.
Like in a scientific fraud and racketeering
enterprise, where Dave Persing's company, Corixa, Imugen and Schoen's L2
Diagnostics are the only labs licensed to use this RICO strain determined to
be burgdorferi via the PrimerShellGame primers - ones not used in any
treatment trials :)
1995;
Seronegative chronic relapsing
neuroborreliosis.
We report an unusual patient with evidence of Borrelia burgdorferi infection who experienced repeated neurologic relapses despite aggressive antibiotic therapy. Each course of therapy was associated with a Jarisch-Herxheimer-like reaction. Although the patient never had detectable free antibodies to B. burgdorferi in serum or spinal fluid, the CSF was positive on multiple occasions for complexed anti-B. burgdorferi antibodies, B. burgdorferi nucleic acids and free antigen.
http://www.ncbi.nlm.nih.gov/pubmed/7796837
1996, Barbour and Bartold on spirochetal
blebbing (auto-vaccination with OspA)
BARBOURS_STEALTH_BOMBERS.htm
The Scientist 1996, 10(14):13
1996, Dec;
Infection by Candida albicans inhibits
apoptosis of human monocytes and
monocytic U937 cells.\
Infectious microorganisms can differently induce or inhibit apoptosis of immunocompetent effector and host cells. In this study we examined the influence of an infection by Candida albicans (C. albicans) on programmed cell death of monocytic U937 cells and human monocytes. Basal and tumor necrosis factor alpha (TNF-alpha)-induced DNA fragmentation of U937 cells was significantly inhibited by an infection with C. albicans. Enhanced apoptosis of U937 cells, induced by TNF-alpha, caused a diminished candidacidal activity of the effector cells, whereas inhibition of apoptosis by granulocyte-macrophage colony-stimulating factor (GM-CSF) was paralleled by an intensified host defense. Pretreatment of U937 cells or monocytes with the cyclooxygenase blocker indomethacin completely abolished the reduction of DNA fragmentation induced by the yeast. Studying the underlying mechanisms we found that C. albicans induced formation of prostaglandin E2 (PGE2) by U937. Exogenous administration of PGE2 down-regulated apoptosis of U937 or human monocytes to a similar extent as did fungal infection. Activation of protein kinase A by the cAMP analogue 8-bromo-cAMP inhibited U937 apoptosis, as did PGE2. On the other hand, rp-cAMP, a blocker of the cAMP-dependent signal transduction, restored and elevated DNA fragmentation levels down-regulated by C. albicans. U937 cells expressed the bcl-2 protein but the infection with fungi or PGE2 treatment did not increase proto-oncogene expression. Monocytic effector cells may therefore strengthen the defense against C. albicans by an autocrine feedback regulation via a PGE2-dependent, cAMP-transduced inhibition of apoptosis.
http://www.ncbi.nlm.nih.gov/pubmed/8975876
1996;
KOREANS : Characterization of Extremely Hydrophobic Immunostimulatory Lipoidal
Peptides by Matrix Assisted Laser Desorption Ionization Mass Spectrometry
Synthetic lipoidal peptides based on viral
protein sequences have been prepared. These peptides contain an N-palmitoyl
group at the N-terminal residue, which is a modified cysteine, containing a
S-[2,3-bis(acyloxy)-(2-R,S)-propyl] moiety. When this residue (Pam3Cys) is
at the N-terminus of a synthetic peptide, it acts as potent immunoadjuvant
to enhance both IgM and IgG antibody responses to the attached peptide.
Conventional analytical procedures (e.g., Edman degradation and amino acid
analysis) are either not applicable due to the N-terminal modification, or
do not provide confirmation of the intact structure. Chromatographic
analysis is also hindered by the tendency of these lipoidal Pam3Cys peptides
to form large aggregates, and in some cases to be permanently adsorbed on
reversed phase columns. We have applied several mass spectrometric
techniques, including fast atom bombardment (FAB), electrospray ionization (ESI)
and matrix assisted laser desorption ionization (MALDI) to characterize the
intact structures of a number of different Pam3Cys synthetic peptides. The
MALDI-MS has been found to be the most sensitive for the analysis of the
structure of Pam3Cys peptides. PDF
1997,
Tully; Mycoplasmas: sophisticated,
reemerging, and burdened by their
notoriety.
Mycoplasmas are most unusual
self-replicating bacteria, possessing very small genomes, lacking cell wall
components, requiring cholesterol for membrane function and growth, using UGA
codon for tryptophan, passing through "bacterial-retaining" filters, and
displaying genetic economy that requires a strict dependence on the host for
nutrients and refuge. In addition, many of the mycoplasmas pathogenic for humans
and animals possess extraordinary specialized tip organelles that mediate their
intimate interaction with eucaryotic cells. This host-adapted survival is
achieved through surface parasitism of target cells, acquisition of essential
biosynthetic precursors, and in some cases, subsequent entry and survival
intracellularly. Misconceptions concerning the role of mycoplasmas in disease
pathogenesis can be directly attributed to their biological subtleties and to
fundamental deficits in understanding their virulence capabilities. In this
review, we highlight the biology and pathogenesis of these procaryotes and
provide new evidence that may lead to increased appreciation of their role as
human pathogens.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2627593/?tool=pubmed
1997, Dave Persing (Lyme Crook)
Molecular evidence and clinical
significance of herpesvirus coinfection in the central nervous system.
A total of 60 cerebrospinal fluid (CSF)
specimens from patients manifesting symptoms resembling viral central
nervous system (CNS) disease were examined for the presence of herpes
simplex virus (HSV), human herpesvirus 6 (HHV-6), Epstein-Barr virus (EBV),
cytomegalovirus, varicella-zoster virus, Borrelia burgdorferi, and
Tropheryma whippelii DNA by PCR. Of 30 specimens which were selected on the
basis of HSV DNA positivity, 2 were concomitantly positive for HHV-6 DNA and
1 was positive for EBV DNA. In the three specimens positive for more than
one herpesvirus, amplicons generated with virus-specific primer sets
hybridized specifically to the corresponding virus-specific probe. Sequence
analysis of the two amplified DNA fragments demonstrated that they were
derived from distinct herpesviruses. Of 22 patients with clinically
diagnosed encephalitis, 2 of 3 patients coinfected with HSV and HHV-6 died,
compared to 1 of 19 (5%) patients infected with only HSV. Of 30 CSF
specimens that were negative for HSV DNA, EBV DNA was detected in one
sample. These data indicated the presence of DNA specific for two distinct
herpesviruses in the same CSF specimen, providing molecular evidence that
coinfection with this group of viruses may occur in the CNS.
In the hyperthermia- and UV light-induced mouse models,
treatment with anti-IL-6 antibodies results in significantly lower
frequencies of ocular reactivation compared with those in mice treated with
a control immunoglobulin (24). Herpesviruses, including HSV, CMV, EBV,
and HHV-6, have been demonstrated to induce a concomitant release of IL-6,
thereby disturbing immune homeostasis (15, 21, 25). This effect may
itself be immunosuppressive, which could allow establishment or reactivation
of other infectious agents within the host, thereby enhancing the occurrence
of coinfection in the CNS.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC230077/pdf/352869.pdf
1997, Aug;
Processing of mycobacterial lipids and
effects on host responsiveness.
One of the most important opportunistic pathogens associated with AIDS is the Mycobacterium avium complex. M. avium infections are found in up to 70% of individuals in advanced stages of AIDS. The deficiency in our knowledge of these mycobacteria presents an obstacle to the development of a rational approach for controlling these life-threatening infections in immunocompromised persons. It is apparent that M. avium can replicate in host macrophages and persist for long periods. During this time, various components, particularly lipids, accumulate in host macrophages and contribute to the ability of this organism to upset the cytokine homeostasis necessary for controlling infections of this type. M. avium lipids are immunosuppressive and can induce a variety of cytokines and eicosanoids that affect general host responses. The intention of this review is to examine the postphagocytic processing of various M. avium lipids with respect to their ability to alter host responses, particularly in immunocompromised patients such as those infected with HIV.
http://www.ncbi.nlm.nih.gov/pubmed/9257946
1997, Sep;
Mycobacterium bovis Bacillus Calmette
Guérin infection prevents apoptosis of
resting human monocytes.
Apoptosis plays an essential role in the development and homeostasis of multicellular organisms. Some infectious agents interfere with this programmed cell death to their own benefit. Here, we show that infection of resting human monocytes with Mycobacterium bovis Bacillus Calmette Guérin (BCG) increases monocyte viability by preventing them from undergoing apoptosis. Heat-killed BCG also prevented apoptosis, indicating that replication of BCG is not required to prevent cell death. Analysis of BCG-infected monocytes revealed an up-regulation of the A1 mRNA, whereas the bcl-2 mRNA was not up-regulated. Interestingly, preinfection with BCG renders the cells resistant to interleukin (IL)-10-induced apoptosis which may be one of the mechanisms mycobacteria use to modulate immune responses. BCG infection was also accompanied by an impairment of the capacity of monocytes to secrete IL-10 and by an induction of the capacity to secrete tumor necrosis factor-alpha, two cytokines known to induce and prevent human monocyte apoptosis, respectively. Since it has been reported that apoptosis is involved in killing of intracellular mycobacteria, the prevention of apoptosis may represent a strategy for mycobacterial survival in the infected host.
http://www.ncbi.nlm.nih.gov/pubmed/9341792?dopt=Abstract
http://onlinelibrary.wiley.com/doi/10.1002/eji.1830270945/abstract
1998;
Lyme Crook Dave Persing does the advertising for the Yale-Imugen-Corixa RICO
cabal
1998, Mark Klempner, OspA-vaccination
results in Anti-Myelin Antibodies:
http://www.actionlyme.org/KFORSCHNER_DISCOVERS_LYME_TOXIN.htm
"Despite
[proven
by Klempner to not ever happen]
appropriate treatment, some patients may
develop ... encephalopathy
[which
Klempner now denies]..."
"In
the central nervous system, T cells that
react to OspA, OspC and p22 also
recognize MOBP [myelin oligodendrocyte
basic protein], SST-R1, and IL- 1R,
respectively, on neurons..."
So, in 1998, ▲Mark Klempner reported
that OspA vaccination could cause a
Multiple Sclerosis outcome.
Now scroll down to Aug, 2010, where
Wormser, Klempner and Latov talk about
how Lyme causes anti-brain antibodies...
This, while the Tribune supports their
bullshit that "Lyme isn't real."
Meanwhile they themselves say
chronic
Lyme and LYMErix Disease is real and
permanent, so WTF??
AUTISM RELATED:
1998, CDC:
"Updated information on adverse events and contraindications, particularly for persons with severe HIV infection, persons with a history of egg allergy or gelatin allergy, persons with a history of thrombocytopenia, and persons receiving steroid therapy [are immune-suppressed]."
http://www.ncbi.nlm.nih.gov/pubmed?term=9639369
1998 May 22;47(RR-8):1-57.Measles, mumps, and rubella--vaccine use and strategies for elimination of measles, rubella, and congenital rubella syndrome and control of mumps: recommendations of the Advisory Committee on Immunization Practices (ACIP).
These revised recommendations of the Advisory Committee on Immunization Practices (ACIP) on measles, mumps, and rubella prevention supersede recommendations published in 1989 and 1990. This statement summarizes the goals and current strategies for measles, rubella, and congenital rubella syndrome (CRS) elimination and for mumps reduction in the United States. Changes from previous recommendations include: Emphasis on the use of combined MMR vaccine for most indications; A change in the recommended age for routine vaccination to 12-15 months for the first dose of MMR, and to 4-6 years for the second dose of MMR; A recommendation that all states take immediate steps to implement a two dose MMR requirement for school entry and any additional measures needed to ensure that all school-aged children are vaccinated with two doses of MMR by 2001; A clarification of the role of serologic screening to determine immunity; A change in the criteria for determining acceptable evidence of rubella immunity; A recommendation that all persons who work in health-care facilities have acceptable evidence of measles and rubella immunity; Changes in the recommended interval between administration of immune globulin and measles vaccination; and Updated information on adverse events and contraindications, particularly for persons with severe HIV infection, persons with a history of egg allergy or gelatin allergy, persons with a history of thrombocytopenia, and persons receiving steroid therapy [are immune-suppressed].
1998,
Philipp:
Borrelia burgdorferi stimulates the
production of interleukin-10 in
peripheral blood mononuclear cells from
uninfected humans and rhesus monkeys.
Heat-killed
Borrelia burgdorferi spirochetes stimulate in vitro production of interleukin-10
(IL-10) at both mRNA and protein levels in peripheral blood mononuclear cells (PBMC)
of uninfected rhesus monkeys. A concomitant down-modulation of IL-2 gene
transcription was observed. Neither IL-4 nor gamma interferon gene expression
was ostensibly affected by B. burgdorferi spirochetes. These phenomena were
observed regardless of whether the stimulating spirochetes belonged to the B.
burgdorferi sensu stricto, Borrelia afzelii, or Borrelia garinii genospecies,
the three main species that cause Lyme disease. B. burgdorferi also induced
production of IL-10 in uninfected human PBMC, indicating that this effect might
play a role in human Lyme disease. Purified lipidated outer surface protein A
(OspA), but not its unlipidated form, induced the production of high levels of
IL-10 in uninfected human PBMC. Thus, the lipid moiety is essential in the
induction of IL-10 in these PBMC. B. burgdorferi M297, a mutant strain that
lacks the plasmid that encodes OspA and OspB, also induced IL-10 gene
transcription in PBMC, indicating that this phenomenon is not causally linked
exclusively to OspA and its lipid moiety. These results demonstrate that B.
burgdorferi can stimulate the production of an antiinflammatory,
immunosuppressive cytokine in naive cells and suggest that IL-10 may play a role
both in avoidance by the spirochete of deleterious immune responses and in
limiting the inflammation that the spirochete is able to induce.
http://www.ncbi.nlm.nih.gov/pubmed/9596735
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC108257/?tool=pubmed
1998;
Detection of Mycoplasma genus and Mycoplasma fermentans by PCR in patients with
Chronic Fatigue Syndrome.
Mycoplasma fermentans and
other Mycoplasma species are colonizers of human mucosal surfaces and may be
associated with human immunodeficiency virus infection. While many infectious
agents have been described in different percentages of patients with Chronic
Fatigue Syndrome (CFS), little is known about the prevalence of mycoplasmas and
especially M. fermentans in CFS patients. A polymerase chain reaction
(PCR)-based assay was used to detect Mycoplasma genus and M. fermentans genomes
in peripheral blood mononuclear cells (PBMC) of CFS patients. Blood was
collected from 100 patients with CFS and 50 control subjects. The amplified
products of 717 bp of Mycoplasma genus, and 206 bp of M. fermentans were
detected in DNA purified from blood samples in 52% and 34% of CFS samples,
respectively. In contrast, these genomes were found in only 14% and 8% of
healthy control subjects respectively (P < 0.0001). All samples were confirmed
by Southern blot with a specific probe based on internal sequences of the
expected amplification product. Several samples, which were positive for
Mycoplasma genus, were negative for M. fermentans indicating that other
Mycoplasma species are involved. A quantitative PCR was developed to determine
the number of M. fermentans genome copies present in 1
microg of DNA for
controls and CFS patients. Mycoplasma copy numbers ranging from 130 to 880 and
from 264 to 2400 were detected in controls and CFS positive subjects,
respectively. An enzyme immunoassay was applied for the detection of antibodies
against p29 surface lipoprotein of M. fermentans to determine the relationship
between M. fermentans genome copy numbers and antibody levels. Individuals with
high genome copy numbers exhibited higher IgG and IgM antibodies against M.
fermentans specific peptides. Isolation of this organism by culture from
clinical specimens is needed in order to demonstrate specificity of signal
detected by PCR in this study.
http://www.ncbi.nlm.nih.gov/pubmed/9879928
SEE ALL RELATED:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=9879928
1999, Jan;
Induction of pro- and anti-inflammatory
cytokines by Borrelia burgdorferi lipoproteins in monocytes is mediated by CD14.
We previously
showed that heat-killed Borrelia burgdorferi spirochetes and lipidated outer
surface protein A (L-OspA) stimulated the in vitro production of
interleukin-10 (IL-10) in peripheral blood mononuclear cells (PBMC) from
uninfected humans and rhesus monkeys (G. Giambartolomei et al., Infect.
Immun. 66:2691-2697, 1998). Here we demonstrate that uninfected human
peripheral blood monocytes, but not B or T cells, are the cells that
transcribe the IL-10 cytokine gene in response to heat-killed B.
burgdorferi. B. burgdorferi similarly induced an upregulation of the
IL-1beta and IL-6 cytokine genes in monocytes and the production of IL-10
and IL-6 in culture supernatants of the human monocytic cell line THP-1.
Purified L-OspA (but not unlipidated OspA [U-OspA] or U-OspC) also
stimulated the production of both cytokines in THP-1 cells in a
dose-dependent fashion, suggesting that acylation of the OspA protein
molecule is required for the production of both anti- and pro-inflammatory
cytokines in naive monocytes. A lipohexapeptide that contained the
tripalmitoyl-modified cysteine motif (Pam3Cys-Hex) of B. burgdorferi
lipoproteins but with an arbitrary peptide sequence had the same effect.
Monoclonal antibodies (MAbs) MY4 and 60bca, both of which bind to CD14 and
are known to block lipopolysaccharide (LPS)-mediated cytokine production,
were able to block L-OspA-mediated IL-10 and IL-6 cytokine production. In
contrast, MAb 26ic, which also binds to CD14 but does not block LPS
function, failed to inhibit L-OspA-mediated cytokine production. These data
suggest that activation of monocytes and production of both anti- and
pro-inflammatory cytokines induced by lipoproteins proceeds via the CD14
receptor. LPS binding protein was not required for OspA-induced cytokine
production. Our results demonstrate that pro- and anti-inflammatory
cytokines induced by B. burgdorferi lipoproteins in PBMC are produced by
monocytes and that lipoprotein and LPS signaling pathways share at least the
initial signaling event that involves the CD14 receptor.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC96289/?tool=pubmed
1999;
Interference by the intracellular parasite Theileria parva with T-cell signal transduction pathways induces
transformation and protection against apoptosis.
The intracellular parasite Theileria parva transforms bovine T-lymphocytes, inducing uncontrolled proliferation. Upon infection, cells cease to require antigenic stimulation and exogenous growth factors to proliferate. Earlier studies have shown that pathways triggered via stimulation of the T-cell receptor are silent in transformed cells. This is reflected by a lack of phosphorylation of key signalling molecules and the fact that proliferation is not inhibited by immunosuppressants such as cyclosporin and ascomycin that target calcineurin. This suggests that the parasite bypasses the normal T-cells activation pathways to induce proliferation. Among the MAP-kinase pathways, ERK and p38 are silent, and only Jun N-terminal kinase is activated. This appears to suffice to induce constitutive activation of the transcription factor AP-1. More recently, it could be shown that the presence of the parasite in the host cell cytoplasm also induces constitutive activation of NF-kappaB, a transcription factor involved in proliferation and protection against apoptosis. Activation is effectuated by parasite-induced degradation of IkappaBs, the cytoplasmic inhibitors which sequester NF-kappaB in the cytoplasm. NF-kappaB activation is resistant to the antioxidant N-acetyl cysteine and a range of other reagents, suggesting that activation might occur in an unorthodox manner. Studies using inhibitors and dominant negative mutants demonstrate that the parasite activates a NF-kappaB-dependent anti-apoptotic mechanism that protects the transformed cell form spontaneous apoptosis and is essential for maintaining the transformed state of the parasitised cell.
http://www.ncbi.nlm.nih.gov/pubmed/10614498
Of course, the next question is do these suckers get transferred by tick?
1999, Radolf, Toll-like receptor 2
functions as a pattern recognition
receptor for diverse bacterial products.
Toll-like receptors (TLRs) 2 and 4 are signal transducers for lipopolysaccharide, the major proinflammatory constituent in the outer membrane of Gram-negative bacteria. We observed that membrane lipoproteins/lipopeptides from Borrelia burgdorferi, Treponema pallidum, and Mycoplasma fermentans activated cells heterologously expressing TLR2 but not those expressing TLR1 or TLR4. These TLR2-expressing cells were also stimulated by living motile B. burgdorferi, suggesting that TLR2 recognition of lipoproteins is relevant to natural Borrelia infection. Importantly, a TLR2 antibody inhibited bacterial lipoprotein/lipopeptide-induced tumor necrosis factor release from human peripheral blood mononuclear cells, and TLR2-null Chinese hamster macrophages were insensitive to lipoprotein/lipopeptide challenge. The data suggest a role for the native protein in cellular activation by these ligands. In addition, TLR2-dependent responses were seen using whole Mycobacterium avium and Staphylococcus aureus, demonstrating that this receptor can function as a signal transducer for a wide spectrum of bacterial products. We conclude that diverse pathogens activate cells through TLR2 and propose that this molecule is a central pattern recognition receptor in host immune responses to microbial invasion.
http://www.ncbi.nlm.nih.gov/pubmed/10559223
1999, Weis;
Cutting edge: inflammatory signaling by Borrelia burgdorferi
lipoproteins is mediated by toll-like receptor 2.
The agent of Lyme disease, Borrelia
burgdorferi, produces membrane lipoproteins possessing potent inflammatory
properties linked to disease pathology. The recent association of toll-like
receptors (TLR) 2 and 4 with LPS responses prompted the examination of TLR
involvement in lipoprotein signaling. The ability of human cell lines to
respond to lipoproteins was correlated with the expression of TLR2.
Transfection of TLR2 into cell lines conferred responsiveness to
lipoproteins, lipopeptides, and sonicated B. burgdorferi, as measured by
nuclear translocation of NF-kappaB and cytokine production. The
physiological importance of this interaction was demonstrated by the 10-fold
greater sensitivity of TLR2-transfected cells to lipoproteins than LPS.
Futhermore, TLR2-dependent signaling by lipoproteins was facilitated by
CD14. These data indicate that TLR2 facilitates the inflammatory events
associated with Lyme arthritis. In addition, the widespread expression of
lipoproteins by other bacterial species suggests that this interaction may
have broad implications in microbial inflammation and pathogenesis.
http://www.ncbi.nlm.nih.gov/pubmed/10452971
http://www.jimmunol.org/cgi/content/full/163/5/2382
1999;
Mycoplasmal infections prevent apoptosis and induce malignant transformation of
interleukin-3-dependent 32D hematopoietic cells.
32D cells, a murine myeloid cell line, rapidly undergo apoptosis upon withdrawal
of interleukin-3 (IL-3) supplement in culture. We found that 32D cells, if
infected by several species of human mycoplasmas that rapidly activated NF-kappaB,
would live and continue to grow in IL-3-depleted culture. Mycoplasma-infected
cells showed no evidence of autocrine production of IL-3. Pyrrolidine
dithiocarbamate (PDTC) blocked activation of NF-kappaB and led to prominent cell
death. Heat-killed mycoplasmas or mycoplasmal membrane preparations alone could
support continued growth of 32D cells in culture without IL-3 supplement for a
substantial period of time. However, upon removal of heat-inactivated
mycoplasmas, 32D cells quickly became apoptotic. In comparison, live Mycoplasma
fermentans or M. penetrans infection for 4 to 5 weeks induced malignant
transformation of 32D cells. Transformed 32D cells grew autonomously and no
longer required support of growth-stimulating factors including IL-3 and
mycoplasmas. The transformed 32D cells quickly formed tumors when injected into
nude mice. Karyotyping showed that development of chromosomal changes and
trisomy 19 was often associated with malignant transformation and tumorigenicity
of 32D cells. Mycoplasmal infections apparently affected the fidelity of genomic
transmission in cell division as well as checkpoints coordinating the
progression of cell cycle events.
http://www.ncbi.nlm.nih.gov/pubmed/10567525
1999,
Sep-Oct;
Lyme Crooks Conflict of Interest Complaint
to "the Government."
Not produced by either Pat Smith or Pam Weintraub
2000; Duray;
Microcystic adnexal carcinoma associated with primary
immunodeficiency, recurrent diffuse herpes simplex virus infection, and
cutaneous T-cell lymphoma.
Cutaneous microcystic adnexal carcinoma
(MAC) is a rare and poorly understood tumor that predominantly occurs in the
head and neck. MAC usually affects people in their fourth and fifth decades.
Some patients have had a history of radiation. We present a case of MAC
occurring in the left antecubital fossa of an 18-year-old white woman with
an unusual immunodeficiency syndrome. The patient also developed a squamous
cell carcinoma, a cutaneous T-cell malignancy, and a perigastric leiomyoma.
A congenital infection of herpes simplex virus (HSV) persisted throughout
her life. The association of HSV infection with MAC and squamous cell
carcinoma and that of peripheral T-cell lymphoma with Epstein-Barr virus is
discussed in relation to her immunodeficiency.
http://www.ncbi.nlm.nih.gov/pubmed/11190445
Congental herpes and primary immunodeficiency: [Use PubMed]
2000, Wormser;
Modulation of lymphocyte proliferative responses by a canine Lyme disease
vaccine of recombinant outer surface protein A (OspA).
"The
modulation of human lymphocyte
proliferative responses was demonstrated
with a recombinant outer surface protein
A (OspA) vaccine preparation for the
prevention of Borrelia burgdorferi
infection. After exposure to either the
unaltered vaccine preparation or OspA
prepared in saline, normal lymphocyte
responses to the mitogens concanavalin
A, phytohemagglutinin-M or pokeweed
mitogen, or the antigen BCG were
consistently reduced. Whole cell
extracts of B. burgdorferi also
modulated immune responses but required
a much greater quantity of protein than
needed for the OspA preparation. The
magnitude of modulation was directly
dependent on the quantity of OspA. OspA
interferes with the response of
lymphocytes to proliferative stimuli
including a blocking of cell cycle phase
progression. Future studies designed to
delete the particular region or
component of the OspA molecule
responsible for this effect may lead to
improved vaccine preparations.
http://www.ncbi.nlm.nih.gov/pubmed/10865170
2000,
Jan; In vitro
Brucella suis infection prevents the
programmed cell death of human monocytic
cells.
"During the complex interaction between an infectious agent and a host organism, the pathogen can interfere with the host cell's programmed death to its own benefit. Induction or prevention of host cell apoptosis appears to be a critical step for determining the infection outcome. Members of the gram-negative bacterial genus Brucella are intracellular pathogens which preferentially invade monocytic cells and develop within these cells. We investigated the effect of Brucella suis infection on apoptosis of human monocytic phagocytes. The present study provides evidence that Brucella infection inhibited spontaneously occurring apoptosis in human monocytes. Prevention of monocyte apoptosis was not mediated by Brucella lipopolysaccharide and required bacterial survival within infected cells. Both invaded and noninvaded cells were protected, indicating that soluble mediators released during infection were involved in the phenomenon. Analysis of Brucella-infected monocytes revealed specific overexpression of the A1 gene, a member of the bcl-2 family implicated in the survival of hematopoietic cells. Brucella infection also rendered macrophage-like cells resistant to Fas ligand- or gamma interferon-induced apoptosis, suggesting that Brucella infection protected host cells from several cytotoxic processes occurring at different steps of the immune response. The present data clearly show that Brucella suis modulated the monocyte/macrophage's apoptotic response to the advantage of the pathogen, thus preventing host cell elimination. This might represent a strategy for Brucella development in infected hosts.
http://www.ncbi.nlm.nih.gov/pubmed/10603407
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC97140/?tool=pubmed
INTRO:
"In recent years, it has become obvious that programmed death (or apoptosis) of cells of the monocytic lineage may be relevant for local immune responses against microorganisms (26, 31). Several bacterial organisms, such as Shigella flexneri (47), Legionella pneumophilia (35), Yersinia enterocolitica (41), Bordetella pertussis (20), Actinobacillus actinomycetemcomitans (18), Listeria monocytogenes (40), and Salmonella enterica serovar Typhimurium (27), promote the destruction of monocytic phagocytes by apoptosis, thus circumventing the first line of defense of the immune system. Surviving bacteria infect neighboring cells and disseminate to other tissues, often epithelial cells. Recently, it was reported that some intracellular bacteria that preferentially infect monocytic phagocytes have a totally opposite strategy and protect their host against apoptosis. Mycobacterium tuberculosis, which was reported to promote alveolar macrophage apoptosis (19, 22), and Mycobacterium bovis were shown to inhibit spontaneously occurring apoptosis in human monocytes (9, 24), possibly by inducing tumor necrosis factor alpha (TNF-α) production. Furthermore, HeLa cells infected with the obligate intracellular bacteria chlamydiae are resistant to apoptosis triggered by exogeneous stimuli (12), and Rickettsia rickettsii prevents the programmed cell death of endothelial cells (7). Molloy et al. showed that apoptosis of BCG-infected macrophages kills intracellular mycobacteria (30). It was thus postulated that inhibition of host cell apoptosis benefits the intracellular pathogen because it protects it against immune attacks from the outside. This could favor an optimal multiplication of intracellular bacteria (7, 9, 12, 24).
2000, May;
The 19-kD antigen and protective immunity in a murine model of
tuberculosis
The 19-kD antigen is a cell wall-associated
lipoprotein present in Mycobacterium tuberculosis and in bacille
Calmette-Guérin (BCG) vaccine strains. Expression of the 19-kD antigen as a
recombinant protein in two saprophytic mycobacteria-M. vaccae and M.
smegmatis-resulted in abrogation of their ability to confer protection
against M. tuberculosis in a murine challenge model, and in their ability to
prime a DTH response to cross-reactive mycobacterial antigens. Induction of
an immune response to the 19-kD antigen by an alternative approach of DNA
vaccination had no effect on subsequent M. tuberculosis challenge. These
results are consistent with a model in which the presence of the 19-kD
protein has a detrimental effect on the efficacy of vaccination with live
mycobacteria. Targeted inactivation of genes encoding selected antigens
represents a potential route towards development of improved vaccine
candidates.
http://www.ncbi.nlm.nih.gov/pubmed/10792376
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1905638/?tool=pubmed
2000, June:
Antiapoptotic herpesvirus Bcl-2 homologs
escape caspase-mediated conversion to
proapoptotic proteins.
The antiapoptotic Bcl-2 and Bcl-x(L) proteins of mammals are converted into potent proapoptotic factors when they are cleaved by caspases, a family of apoptosis-inducing proteases (E. H.-Y. Cheng, D. G. Kirsch, R. J. Clem, R. Ravi, M. B. Kastan, A. Bedi, K. Ueno, and J. M. Hardwick, Science 278:1966-1968, 1997; R. J. Clem, E. H.-Y. Cheng, C. L. Karp, D. G. Kirsch, K. Ueno, A. Takahashi, M. B. Kastan, D. E. Griffin, W. C. Earnshaw, M. A. Veliuona, and J. M. Hardwick, Proc. Natl. Acad. Sci. USA 95:554-559, 1998). Gamma herpesviruses also encode homologs of the Bcl-2 family. All tested herpesvirus Bcl-2 homologs possess antiapoptotic activity, including the more distantly related homologs encoded by murine gammaherpesvirus 68 (gammaHV68) and bovine herpesvirus 4 (BHV4), as described here. To determine if viral Bcl-2 proteins can be converted into death factors, similar to their cellular counterparts, five herpesvirus Bcl-2 homologs from five different viruses were tested for their susceptibility to caspases. Only the viral Bcl-2 protein encoded by gammaHV68 was susceptible to caspase digestion. However, unlike the caspase cleavage products of cellular Bcl-2, Bcl-x(L), and Bid, which are potent inducers of apoptosis, the cleavage product of gammaHV68 Bcl-2 lacked proapoptotic activity. KSBcl-2, encoded by the Kaposi's sarcoma-associated herpesvirus, was the only viral Bcl-2 homolog that was capable of killing cells when expressed as an N-terminal truncation. However, because KSBcl-2 was not cleavable by caspases, the latent proapoptotic activity of KSBcl-2 apparently cannot be released. The Bcl-2 homologs encoded by herpesvirus saimiri, Epstein-Barr virus, and BHV4 were not cleaved by apoptotic cell extracts and did not possess latent proapoptotic activities. Thus, herpesvirus Bcl-2 homologs escape negative regulation by retaining their antiapoptotic activities and/or failing to be converted into proapoptotic proteins by caspases during programmed cell death.
http://www.ncbi.nlm.nih.gov/pubmed/10799576
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC110854/?tool=pubmed
2000;
Mycoplasmal infections alter gene expression in cultured human prostatic and cervical
epithelial cells.
To
better understand how infections by mycoplasmas affect gene expression in human
cells, we quantitatively measured the transcripts of 38 cytokine genes in HPV
E6- and E7-immortalized cervical and prostatic epithelial cells before and after
infection by four human urogenital mycoplasmas, M. fermentans, M. genitalium, M.
hominis and M. penetrans. Using the multi-probe RNase protection assay (RPA), 22
and 23 cytokine gene transcripts were detected in the non-infected control
prostatic and cervical epithelial cells, respectively. Although there were no
discernible changes in cell morphology and growth kinetics following 72 h of
mycoplasmal infection, 55-74% of the cytokine genes expressed in the two human
epithelial cell lines were altered. Most changes reflected an increased
expression of these cytokine genes, while expression of some cytokine genes
significantly decreased. The effects varied with host cell type and species of
infecting mycoplasmas. These alterations in gene expression were more profound
in the cervical epithelial cells than in the prostatic cells. M. fermentans
produced the most significant effects, followed by M. penetrans, M. genitalium
and M. hominis. Some alterations in the gene expression were transient, but most
persisted over the course of chronic (9 months) mycoplasmal infection. Prolonged
gene expression changes induced by chronic mycoplasmal infection may gradually
alter important biological properties in the infected mammalian cells and
produce a unique form of disease process.
http://www.ncbi.nlm.nih.gov/pubmed/10617789
2000;
Polymerase chain reaction for the laboratory diagnosis of aseptic
meningitis and encephalitis.
A protocol for testing cerebrospinal fluid
specimens using a range of PCR assays for the diagnosis of central nervous
system infection was developed and used to test prospectively 383 specimens.
PCR assays were used for the detection of adenovirus, Borrelia burgdorferi,
enteroviruses, Epstein Barr virus, cytomegalovirus, herpes simplex virus,
human herpes virus type 6, JC virus, Leptospira interrogans, Listeria
monocytogenes, lymphocytic choriomeningitis virus, measles virus, mumps
virus, Mycobacterium sp. , Mycoplasma pneumoniae, Toxoplasma gondii and
varicella zoster virus. Of the 383 specimens tested in this study, 46
(12.0%) were found to be positive. The microorganisms detected were CMV,
enterovirus, Epstein Barr virus, herpes simplex virus, human herpes virus
type 6, JC virus, L. monocytogenes, Mycobacterium genus, Toxoplasma gondii
and varicella zoster virus. The introduction of the PCR protocol described
has improved the diagnosis of a range of central nervous system infections
in our laboratory. We believe however that further evaluation of these
assays in immunocompromised patients is necessary to better determine the
predictive value of positive PCR results in these patient groups. http://www.ncbi.nlm.nih.gov/pubmed/11018820
2000, Nov; A novel polymorphism in the
toll-like receptor 2 gene and its
potential association with
staphylococcal infection.
The toll-like receptor 2 (TLR2) has gained importance as a major mammalian receptor for lipoproteins derived from the cell wall of a variety of bacteria, such as Borrelia burgdorferi, Treponema pallidum, and Mycoplasma fermentans. We were interested in identifying mutations in the TLR2 gene that might prove to be associated with altered susceptibility to septic shock. We performed a mutation screen of the TLR2 gene using single-stranded conformational polymorphism in 110 normal, healthy study subjects and detected an Arg753Gln mutation in three individuals. No other missense mutations were detected in the TLR2 open reading frame. Functional studies demonstrate that the Arg753Gln polymorphism, in comparison to the wild-type TLR2 gene, is significantly less responsive to bacterial peptides derived from B. burgdorferi and T. pallidum. In a septic shock population, the Arg753Gln TLR2 polymorphism occurred in 2 out of 91 septic patients. More importantly, both of the subjects with the TLR2 Arg753Gln polymorphism had staphylococcal infections. These findings suggest that a mutation in the TLR2 gene may predispose individuals to life-threatening bacterial infections.
http://www.ncbi.nlm.nih.gov/pubmed/11035751
2000, Dec;
Mycobacterial 19-kDa lipoprotein
mediates Mycobacterium
tuberculosis-induced apoptosis in
monocytes/macrophages at early stages of
infection.
Figure 1C shows that the pre-opsonization of bacilli with anti-19 kDa monoclonal antibody resulted in a strong inhibition of MTB-induced apoptosis (Figure 1C, filled histogram, 21% of Annexin V positive cells).
http://www.nature.com/cdd/journal/v7/n12/full/4400761a.html
In fact, previously reported results showed that low levels of MTB (13) or BCG (15) prevent monocyte apoptosis whereas high amounts of the virulent strain of MTB H37Rv can induce apoptosis.2,3 It is plausible, therefore, to hypothesize that the presence of a threshold signal, represented by the amount of mycobacteria and possibly by the amount of cell wall associated 19-kDa lipoprotein, determines whether the cell will live or die.
13. Durrbaum-Landmann I et al. (1996) Infect. Immun. 64: 5384 ± 5389
15. Kremer L et al. (1997) Eur. J. Immunol. 27: 2450 - 2456
http://www.ncbi.nlm.nih.gov/pubmed/11270362
http://www.nature.com/cdd/journal/v7/n12/pdf/4400761a.pdf
2001,
[Radolf] Toll-like
receptor 2-dependent inhibition of macrophage class II MHC expression and
antigen processing by 19-kDa lipoprotein of Mycobacterium tuberculosis.
Mycobacterium tuberculosis (MTB) induces
vigorous immune responses, yet persists inside macrophages, evading host
immunity. MTB bacilli or lysate was found to inhibit macrophage expression
of class II MHC (MHC-II) molecules and MHC-II Ag processing. This report
characterizes and identifies a specific component of MTB that mediates these
inhibitory effects. The inhibitor was extracted from MTB lysate with Triton
X-114, isolated by gel electroelution, and identified with Abs to be MTB
19-kDa lipoprotein. Electroelution- or immunoaffinity-purified MTB 19-kDa
lipoprotein inhibited MHC-II expression and processing of both soluble Ags
and Ag 85B from intact MTB bacilli. Inhibition of MHC-II Ag processing by
either MTB bacilli or purified MTB 19-kDa lipoprotein was dependent on
Toll-like receptor (TLR) 2 and independent of TLR 4. Synthetic analogs of
lipopeptides from Treponema pallidum also inhibited Ag processing.
Despite the ability of MTB 19-kDa lipoprotein to activate microbicidal and
innate immune functions early in infection, TLR 2-dependent inhibition of
MHC-II expression and Ag processing by MTB 19-kDa lipoprotein during later
phases of macrophage infection may prevent presentation of MTB Ags and
decrease recognition by T cells. This mechanism may allow intracellular MTB
to evade immune surveillance and maintain chronic infection.
http://www.ncbi.nlm.nih.gov/pubmed/11441098
”Synthetic analogs of T.
pallidum,” by Justin Radolf (See 1990, above)
2001;
Mycobacterium tuberculosis 19-kilodalton lipoprotein inhibits
Mycobacterium smegmatis-induced cytokine production by human macrophages in
vitro.
Vaccination of mice with Mycobacterium
vaccae or M. smegmatis induces some protection against M. tuberculosis
challenge. The 19-kDa lipoprotein of M. tuberculosis, expressed in M. vaccae
or M. smegmatis (M. smeg19kDa), abrogates this protective immunity. To
investigate the mechanism of this suppression of immunity, human monocyte-derived
macrophages (MDM) were infected with M. smeg19kDa. Infection resulted in
reduced production of tumor necrosis factor alpha (TNF-alpha) (P < 0.01),
interleukin-12 (IL-12) (P < 0.05), IL-6 (P < 0.05), and IL-10 (P < 0.05),
compared to infection with M. smegmatis vector (M. smegV). Infection with M.
smeg19kDa and with M. smegV had no differential effect on expression of
costimulatory molecules on MDM, nor did it affect the proliferation of
presensitized T cells cocultured with infected MDM. When MDM were infected
with M. smegmatis expressing mutated forms of the 19-kDa lipoprotein,
including non-O-glycosylated (M. smeg19NOG), nonsecreted (M. smeg19NS), and
nonacylated (M. smeg19NA) variants, the reduced production of TNF-alpha or
IL-12 was not observed. When the purified 19-kDa lipoprotein was added
directly to cultures of infected monocytes, there was little effect on
either induction of cytokine production or its inhibition. Thus, the
immunosuppressive effect is dependent on glycosylated and acylated 19-kDa
lipoprotein present in the phagosome containing the mycobacterium. These
results suggest that the diminished protection against challenge with M.
tuberculosis seen in mice vaccinated with M. smegmatis expressing the 19-kDa
lipoprotein is the result of reduced TNF-alpha and IL-12 production,
possibly leading to reduced induction of T-cell activation.
http://www.ncbi.nlm.nih.gov/pubmed/11179309
2001;
Induction of tolerance to lipopolysaccharide and
mycobacterial components in Chinese hamster ovary/CD14 cells is not affected by overexpression of Toll-like receptors 2 or 4.
Down-regulation of cell surface expression of Toll-like receptor (TLR) 4
following LPS stimulation has been suggested to underlie endotoxin tolerance. In
this study, we examined whether overexpression of TLR2 or TLR4 would affect the
ability of cells to become tolerant to LPS or the mycobacterial components,
arabinose-capped lipoarabinomannan (LAM) and soluble tuberculosis factor (STF).
To this end, Chinese hamster ovary/CD14 cells stably transfected with a NF-kappaB-dependent
reporter construct, endothelial leukocyte adhesion molecule CD25 (the 3E10
clone), were engineered to overexpress either human TLR2 or TLR4. Transfected
TLRs exhibited proper signaling functions, as evidenced by increased LPS
responsiveness of 3E10/TLR4 cells and acquisition of sensitivity to
TLR2-specific ligands upon transfection of TLR2 into TLR2-negative 3E10 cells.
Pretreatment of cells with LPS, LAM, or STF did not modulate TLR2 or TLR4 cell
surface expression. Following LPS exposure, 3E10, 3E10/TLR2, and 3E10/TLR4 cells
exhibited comparable decreases in LPS-mediated NF-kappaB activation and mitogen-activated
protein (MAP) kinase phosphorylation. Likewise, LPS pretreatment profoundly
inhibited LPS-induced NF-kappaB translocation in Chinese hamster ovary cells
that concomitantly overexpressed human TLR4 and myeloid differentiation
protein-2 (MD-2), but failed to modulate TLR4 or MD-2 cell surface expression.
Pretreatment of 3E10/TLR2 cells with LAM or STF decreased their NF-kappaB
responses induced by subsequent stimulation with these substances or LPS.
Conversely, prior exposure of 3E10/TLR2 cells to LPS led to hyporesponsiveness
to LPS, LAM, and STF, indicating that LPS and mycobacterial products induce
cross-tolerance. Thus, tolerance to LPS and mycobacterial components cannot be
attributed solely to a decrease in TLR/MD-2 expression levels, suggesting
inhibition of expression or function of other signaling intermediates 2002,
Induction of bacterial lipoprotein tolerance is associated with suppression of
toll-like receptor 2 expression.
http://www.ncbi.nlm.nih.gov/pubmed/11490013
2002, Sep,
Nicolson;
High frequency of systemic
mycoplasmal infections in Gulf War veterans and civilians with Amyotrophic
Lateral Sclerosis (ALS).
The presence of systemic mycoplasmal
infections in the blood of Gulf War veterans (n=8) and civilians (n=28) with
Amyotrophic Lateral Sclerosis (ALS) and age matched controls (n=70) was
investigated by detecting mycoplasma gene sequences with forensic Polymerase
Chain Reaction (PCR) and back hybridization with a radiolabeled internal
oligonucleotide probe. Almost all ALS patients (30/36 or approximately 83%)
showed evidence of Mycoplasma species in blood samples, whereas <9% of
controls had blood mycoplasmal infections (P<0.001). Using PCR ALS patients
with a positive test for any mycoplasmal infection were investigated for the
presence of M. fermentans, M. pneumoniae, M. hominis and M. penetrans in
their blood. All Gulf War veterans with ALS were positive for M. fermentans,
except one that was positive for M. genitalium. In contrast, the 22/28
civilians with detectable mycoplasmal infections had M. fermentans (13/22,
59%) as well as other Mycoplasama species in their blood, and two of the
civilian ALS patients had multiple mycoplasma species (M. fermentans plus M.
hominis). Of the few control patients that were positive, only two patients
(2/70, 2.8%) were positive for M. fermentans (P<0.001). The results support
the suggestion that infectious agents may play a role in the pathogenesis
and/or progression of ALS, or alternatively ALS patients are extremely
susceptible to systemic mycoplasmal infections.
http://www.ncbi.nlm.nih.gov/pubmed/12383408
Full Text:
http://www.actionlyme.org/GARTHNICOLSON.pdf
Excerpt:
"Mycoplasmas like M. fermentans are particularly interesting because they have the capacity, like emnteroviruses 7,11 to penetrate the CNS, and they possess the potential tocause oersistent neurolohical signs and symptoms. 16-18 Our results on mycoplasmal infecttions in ALS patients suggests that the coinfections with certain persistent viruses might be important in ALS. We also found that ALS patients have some of the signs and symptoms seen in a variety of chronic illness patients, consistent with having mycoplasmal infections that are also found in these patients. 18-20k 28, 29 Similar to chronic mycoplasmal infections, 21-23, 27-29 enteroviruses have also been found in patients with chronic myocarditis 30 [and one would think, chronic infections of the nerves and nerve roots, like FibroFemino-Not-Enough-Sexitis--- KMD] and chronic fatigu. 30 It is interesting that both enteroviruses have the capacity to cause slow, persistent infetiona that can eventually result in cellular dysfunction and eventually cell death, 11, 29, 32 and mycoplasmal infections have been implicated in autoimmune diseases, 17, 18, 20, 28, 29, 32 and one hypothesis on the pathogenisis of ALS suggests an autoimmune involvement. 33 "
2002,
Donta; The antibiotic treatment trial of Gulf War
Veterans' Illnesses: issues, design, screening, and baseline characteristics.
Many veterans who were deployed to the
Persian Gulf during the 1990-1991 Gulf War developed multiple unexplained
symptoms such as pain, fatigue, and neurocognitive problems. This
constellation of symptoms has been termed Gulf War Veterans' Illnesses (GWVI).
Although there is no proven explanation for the cause of GWVI, one fairly
widespread explanation is systemic Mycoplasma fermentans infection. The
Antibiotic Treatment Trial of GWVI is a randomized placebo-controlled trial
to determine whether a 1-year course of doxycycline treatment in deployed
Gulf War veterans with GWVI and testing as Mycoplasma species positive will
improve their overall functional status as measured by the Physical
Component Summary of the SF-36V questionnaire. The study of a multisymptom
illness such as GWVI is complicated by the nonspecific nature of the
illness, the unknown etiology, and the lack of a widely accepted outcome
measure. The presumption of mycoplasma infection raises concerns
regarding the methodology for determination of mycoplasma infection, the
choice of treatment, and the duration of treatment. However, such a
presumption allows the formulation of a clear testable hypothesis that can
be tested with treatments with known rates of adverse events and known
activity against Mycoplasma species. This paper describes the major
issues faced by the investigators during planning, the study design, the
patient screening results, and the baseline characteristics of the study
patients. There were 2712 patients screened for study entry at 26 Department
of Veterans Affairs and two Department of Defense medical centers. Of these,
491 met all study entry criteria and were randomized to either 1 year of
doxycycline (200 mg/day) or 1 year of placebo. All patients were seen
monthly during treatment and at 6 months after the end of treatment. Study
patients had a mean age of 41 years and were mostly male (86%), white (64%),
married (68%), and employed full-time (71%).
http://www.ncbi.nlm.nih.gov/pubmed/12057884
This study was
incomplete due to non-compliance.
2002;
Induction of bacterial lipoprotein tolerance is associated with suppression of
toll-like receptor 2 expression.
Tolerance to bacterial
cell wall components including lipopolysaccharide (LPS) may represent an
essential regulatory mechanism during bacterial infection. Two members of
the Toll-like receptor (TLR) family, TLR2 and TLR4, recognize the specific
pattern of bacterial cell wall components. TLR4 has been found to be
responsible for LPS tolerance. However, the role of TLR2 in bacterial
lipoprotein (BLP) tolerance and LPS tolerance is unclear. Pretreatment of
human THP-1 monocytic cells with a synthetic bacterial lipopeptide induced
tolerance to a second BLP challenge with diminished tumor necrosis
factor-alpha and interleukin-6 production, termed BLP tolerance.
Furthermore, BLP-tolerized THP-1 cells no longer responded to LPS
stimulation, indicating a cross-tolerance to LPS. Induction of BLP tolerance
was CD14-independent, as THP-1 cells that lack membrane-bound CD14 developed
tolerance both in serum-free conditions and in the presence of a specific
CD14 blocking monoclonal antibody (MEM-18). Pre-exposure of THP-1 cells to
BLP suppressed mitogen-activated protein kinase phosphorylation and nuclear
factor-kappaB activation in response to subsequent BLP and LPS stimulation,
which is comparable with that found in LPS-tolerized cells, indicating that
BLP tolerance and LPS tolerance may share similar intracellular pathways.
However, BLP strongly enhanced TLR2 expression in non-tolerized THP-1 cells,
whereas LPS stimulation had no effect. Furthermore, a specific TLR2 blocking
monoclonal antibody (2392) attenuated BLP-induced, but not LPS-induced,
tumor necrosis factor-alpha and interleukin-6 production, indicating BLP
rather than LPS as a ligand for TLR2 engagement and activation. More
importantly, pretreatment of THP-1 cells with BLP strongly inhibited TLR2
activation in response to subsequent BLP stimulation. In contrast, LPS
tolerance did not prevent BLP-induced TLR2 overexpression. These results
demonstrate that BLP tolerance develops through down-regulation of TLR2
expression.
http://www.ncbi.nlm.nih.gov/pubmed/12133836
2002;
Lipopolysaccharide-
and lipoteichoic acid-induced tolerance and cross-tolerance: distinct
alterations in IL-1 receptor-associated kinase.
Human Toll-like
receptor (TLR) 4 and TLR2 receptors recognize LPS or lipoteichoic acid (LTA),
respectively. Prolonged exposure of human macrophages/monocytes to bacterial LPS
induces a state of adaptation/tolerance to subsequent LPS challenge.
Inflammatory gene expressions such as IL-1beta and TNF-alpha are selectively
repressed, while certain anti-inflammatory genes such as secretory IL-1R
antagonist are still induced in LPS-adapted/tolerant cells. In this report, we
demonstrate that LPS-tolerized human promonocytic THP-1 cells develop
cross-tolerance and no longer respond to LTA-induced IL-1beta/TNF-alpha
production, indicating that disruption of common intracellular signaling is
responsible for the decreased IL-1beta/TNF-alpha production. We observe that
down-regulation of IL-1R-associated kinase (IRAK) protein level and kinase
activity closely correlates with the development of cross-tolerance. IRAK
protein levels and kinase activities in LPS-tolerized cells remain low and
hyporesponsive to subsequent LPS or LTA challenges. We also demonstrate that
THP-1 cells with prolonged LTA treatment develop LTA tolerance and do not
express IL-1beta/TNF-alpha upon further LTA challenge. Strikingly, cells
tolerized with LTA are only refractory to subsequent LTA challenge and can still
respond to LPS stimulation. Correspondingly, stimulation of TLR2 by LTA,
although activating IRAK, does not cause IRAK degradation. IRAK from
LTA-tolerized cells can be subsequently activated and degraded by further LPS
challenge, but not LTA treatment. Our studies reveal that LTA-induced tolerance
is distinct compared with that of LPS tolerance, and is likely due to disruption
of unique TLR2 signaling components upstream of MyD88/IRAK.
http://www.ncbi.nlm.nih.gov/pubmed/12055225
http://www.jimmunol.org/cgi/content/full/168/12/6136
2003,
Hulinska,
Interaction of Borrelia burgdorferi
sensu lato with Epstein-Barr virus in
lymphoblastoid cells.
Since the possibility of interruption of latent EBV infection has been suggested
by the induction of the lytic virus cycle with chemical substances, other
viruses, and by immunosuppression, we hypothesized that the same effect might
happen in B. burgdorferi sensu lato infection as happens in Lyme disease
patients with positive serology for both agents. We have observed EBV
replication in lymphoblastoid cells after superinfection with B. garinii and B.
afzelii strains after 1 and 4 h of their interaction. We found that viral and
borrelial antigens persisted in the lymphoblasts for 3 and 4 days. Morphological
and functional transformation of both agents facilitate their transfer to
daughter cells. Association with lymphoblasts and internalization of B. garinii
by tube phagocytosis increased replication of viruses more successfully than B.
afzelii and chemical inductors. Demonstration of such findings must be
interpreted cautiously, but may prove a mixed borrelial and viral cause of
severe neurological disease.
http://www.ncbi.nlm.nih.gov/pubmed/12630667
2003;
Borrelia burgdorferi-induced
tolerance as a model of persistence via immunosuppression.
If left untreated, infection with Borrelia
burgdorferi sensu lato may lead to chronic Lyme borreliosis. It is still
unknown how this pathogen manages to persist in the host in the presence of
competent immune cells. It was recently reported that Borrelia suppresses
the host's immune response, thus perhaps preventing the elimination of the
pathogen (I. Diterich, L. Härter, D. Hassler, A. Wendel, and T. Hartung,
Infect. Immun. 69:687-694, 2001). Here, we further characterize
Borrelia-induced immunomodulation in order to develop a model of this anergy.
We observed that the different Borrelia preparations that we tested, i.e.,
live, heat-inactivated, and sonicated Borrelia, could desensitize human
blood monocytes, as shown by attenuated cytokine release upon restimulation
with any of the different preparations. Next, we investigated whether these
Borrelia-specific stimuli render monocytes tolerant, i.e. hyporesponsive,
towards another Toll-like receptor 2 (TLR2) agonist, such as lipoteichoic
acid from gram-positive bacteria, or towards the TLR4 agonist
lipopolysaccharide. Cross-tolerance towards all tested stimuli was induced.
Furthermore, using primary bone marrow cells from TLR2-deficient mice and
from mice with a nonfunctional TLR4 (strain C3H/HeJ), we demonstrated that
the TLR2 was required for tolerance induction by Borrelia, and using
neutralizing antibodies, we identified interleukin-10 as the key mediator
involved. Although peripheral blood mononuclear cells tolerized by Borrelia
exhibited reduced TLR2 and TLR4 mRNA levels, the expression of the
respective proteins on monocytes was not decreased, ruling out the
possibility that tolerance to Borrelia is attributed to a reduced TLR2
expression. In summary, we characterized tolerance induced by B.
burgdorferi, describing a model of desensitization which might mirror the
immunosuppression recently attributed to the persistence of Borrelia in
immunocompetent hosts.
http://www.ncbi.nlm.nih.gov/pubmed/12819085
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC162029/?tool=pubmed
2003, Jun;
The Mycobacterium
tuberculosis recombinant 27-kilodalton lipoprotein induces a strong Th1-type
immune response deleterious to protection.
Th1 immune response is essential in the
protection against mycobacterial intracellular pathogens. Lipoproteins
trigger both humoral and cellular immune responses and may be candidate
protective antigens. We studied in BALB/c mice the immunogenicity and the
protection offered by the recombinant 27-kDa Mycobacterium tuberculosis
lipoprotein and the corresponding DNA vaccine. Immunization with the 27-kDa
antigen resulted in high titers of immunoglobulin G1 (IgG1) and IgG2a with a
typical Th1 profile and a strong delayed hypersensitivity response. A strong
proliferation response was observed in splenocytes, and significant nitric
oxide production and gamma interferon secretion but not interleukin 10
secretion were measured. Based on these criteria, the 27-kDa antigen induced
a typical Th1-type immune response thought to be necessary for protection.
Surprisingly, in 27-kDa-vaccinated mice (protein or DNA vaccines)
challenged by M. tuberculosis H37Rv or BCG strains, there was a significant
increase in the numbers of CFU in the spleen compared to that for control
groups. Furthermore, the protection provided by BCG or other mycobacterial
antigens was completely abolished once the 27-kDa antigen was added to the
vaccine preparations. This study indicates that the 27-kDa antigen has an
adverse effect on the protection afforded by recognized vaccines. We are
currently studying how the 27-kDa antigen modulates the mouse immune
response.
http://www.ncbi.nlm.nih.gov/pubmed/12761093
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC155707/?tool=pubmed
2003, Nicolson; Multiple co-infections
(Mycoplasma, Chlamydia, human herpes
virus-6) in blood of chronic fatigue
syndrome patients: association with
signs and symptoms.
Previously we and others found that a majority of chronic fatigue syndrome (CFS) patients showed evidence of systemic mycoplasmal infections, and their blood tested positive using a polymerase chain reaction assay for at least one of the four following Mycoplasma species: M. fermentans, M. hominis, M. pneumoniae or M. penetrans. Consistent with previous results, patients in the current study (n=200) showed a high prevalence (overall 52%) of mycoplasmal infections. Using forensic polymerase chain reaction we also examined whether these same patients showed evidence of infections with Chlamydia pneumoniae (overall 7.5% positive) and/or active human herpes virus-6 (HHV-6, overall 30.5% positive). Since the presence of one or more infections may predispose patients to other infections, we examined the prevalence of C. pneumoniae and HHV-6 active infections in mycoplasma-positive and -negative patients. Unexpectedly, we found that the incidence of C. pneumoniae or HHV-6 was similar in Mycoplasma-positive and -negative patients, and the converse was also found in active HHV-6-positive and -negative patients. Control subjects (n=100) had low rates of mycoplasmal (6%), active HHV-6 (9%) or chlamydial (1%) infections, and there were no co-infections in control subjects. Differences in bacterial and/or viral infections in CFS patients compared to control subjects were significant. Severity and incidence of patients' signs and symptoms were compared within the above groups. Although there was a tendency for patients with multiple infections to have more severe signs and symptoms (p<0.01), the only significant differences found were in the incidence and severity of certain signs and symptoms in patients with multiple co-infections of any type compared to the other groups (p<0.01). There was no correlation between the type of co-infection and severity of signs and symptoms. The results indicate that a large subset of CFS patients show evidence of bacterial and/or viral infection(s), and these infections
may contribute to the severity of signs and symptoms found in these patients.
http://www.ncbi.nlm.nih.gov/pubmed/12887507
2003, July, Anthony Fauci says on
CNN (Lou
Dobbs)
"We've had an effective vaccine,
but it's the kind of vaccine that you
have to essentially vaccinate people
each year. And from the standpoint of
it's use, it has not been used as
efficiently as it could have been used.
So scientifically, we had a vaccine and
still do have a vaccine, but it's not
really well used." -- Fauci
...because fauci is a brainless tard and unable to learn new things. It took 5 years and a failed huge HIV trial for him to figger out you can't use Pam3Cys as a vaccine.
2003,
Induction of In
Vitro Reprogramming by Toll-Like Receptor (TLR)2 and TLR4 Agonists in Murine
Macrophages: Effects of TLR "Homotolerance" Versus "Heterotolerance" on NF-B
Signaling Pathway Components1
In this study, tolerance
induction by preexposure of murine macrophages to Toll-like receptor (TLR)2 and
TLR4 agonists was revisited, focusing on the major signaling components
associated with NF-kappaB activation. Pretreatment of macrophages with a pure
TLR4 agonist (protein-free Escherichia coli (Ec) LPS) or with TLR2 agonists
(Porphyromonas gingivalis LPS or synthetic lipoprotein Pam3Cys) led to
suppression of TNF-alpha secretion, IL-1R-associated kinase-1, and IkappaB
kinase (IKK) kinase activities, c-jun N-terminal kinase, and extracellular
signal-regulated kinase phosphorylation, and to suppression of NF-kappaB DNA
binding and transactivation upon challenge with the same agonist (TLR4 or TLR2
"homotolerance," respectively). Despite inhibited NF-kappaB DNA binding,
increased levels of nuclear NF-kappaB were detected in agonist-pretreated
macrophages. For all the intermediate signaling elements, heterotolerance was
weaker than TLR4 or TLR2 homotolerance with the exception of IKK kinase
activity. IKK kinase activity was unperturbed in heterotolerance. TNF-alpha
secretion was also suppressed in P. gingivalis LPS-pretreated, Ec LPS-challenged
cells, but not vice versa, while Pam3Cys and Ec LPS did not induce a state of
cross-tolerance at the level of TNF-alpha. Experiments designed to elucidate
novel mechanisms of NF-kappaB inhibition in tolerized cells revealed the
potential contribution of IkappaBepsilon and IkappaBxi inhibitory proteins and
the necessity of TLR4 engagement for induction of tolerance to Toll
receptor-IL-1R domain-containing adapter protein/MyD88-adapter-like-dependent
gene expression. Collectively, these data
demonstrate that induction of homotolerance affects a broader spectrum of
signaling components than in heterotolerance, with selective modulation of
specific elements within the NF-kappaB signaling pathway.
http://www.ncbi.nlm.nih.gov/pubmed/12496438
http://www.jimmunol.org/cgi/content/full/170/1/508
2004:
Mycobacterium
tuberculosis LprG (Rv1411c): A Novel TLR-2 Ligand That Inhibits Human
Macrophage Class II MHC Antigen Processing1
"Signaling
through TLR-2 by lipoproteins may represent a double-edged sword for host responses to chronic intracellular pathogens such as M. tuberculosis. Short-term signaling through TLR-2 activates macrophages and initiates acute inflammation that may help control initial infection. In contrast, prolonged
TLR-2 signaling in macrophages results in down-regulation of
certain critical immune functions, such as MHC-II Ag
processing. M. tuberculosis infects, survives, and
persists in macrophages. The ability of M. tuberculosis to
survive acute inflammation positions the bacilli to take
advantage, through secretion of lipoproteins such as LprG and
LpqH, of this
down-regulation of macrophage immune function."
http://www.jimmunol.org/cgi/content/full/173/4/2660
2004;
Mycobacterium tuberculosis inhibits macrophage responses to IFN-gamma
through myeloid differentiation factor 88-dependent and -independent mechanisms.
Mycobacterium tuberculosis overcomes
macrophage bactericidal activities and persists intracellularly. One
mechanism by which M. tuberculosis avoids macrophage killing might be
through inhibition of IFN-gamma-mediated signaling. In this study we provide
evidence that at least two distinct components of M. tuberculosis, the
19-kDa lipoprotein and cell wall peptidoglycan (contained in the
mycolylarabinogalactan peptidoglycan (mAGP) complex), inhibit macrophage
responses to IFN-gamma at a transcriptional level. Moreover, these
components engage distinct proximal signaling pathways to inhibit responses
to IFN-gamma: the 19-kDa lipoprotein inhibits IFN-gamma signaling in a
Toll-like receptor (TLR)2-dependent and myeloid differentiation factor
88-dependent fashion whereas mAGP inhibits independently of TLR2, TLR4, and
myeloid differentiation factor 88. In addition to inhibiting the induction
of specific IFN-gamma responsive genes, the 19-kDa lipoprotein and mAGP
inhibit the ability of IFN-gamma to activate murine macrophages to kill
virulent M. tuberculosis without inhibiting production of NO. These results
imply that inhibition of macrophage responses to IFN-gamma may contribute to
the inability of an apparently effective immune response to eradicate M.
tuberculosis.
http://www.ncbi.nlm.nih.gov/pubmed/15128816
2004, Sep, Latov (See
LATOV_GAMMOPATHY.htm) ; Neuropathy
and cognitive impairment following
vaccination with the OspA protein of
Borrelia burgdorferi.
Neurological syndromes that follow vaccination or infection are often attributed to autoimmune mechanisms. We report six patients who developed neuropathy or cognitive impairment, within several days to 2 months, following vaccination with the OspA antigen of Borrelia burgdorferi. Two of the patients developed cognitive impairment, one chronic inflammatory demyelinating polyneuropathy (CIDP), one multifocal motor neuropathy, one both cognitive impairment and CIDP, and one cognitive impairment and sensory axonal neuropathy. The patients with cognitive impairment had T2 hyperintense white matter lesions on magnetic resonance imaging. The similarity between the neurological sequelae observed in the OspA-vaccinated patients and those with chronic Lyme disease suggests a possible role for immune mechanisms in some of the manifestations of chronic Lyme disease that are resistant to antibiotic treatment.
http://www.ncbi.nlm.nih.gov/pubmed/15363064
2004; Philipp;
Lipoproteins, not
lipopolysaccharide, are the key mediators of the proinflammatory response elicited by heat-killed
Brucella abortus.
Inflammation is a hallmark of brucellosis.
Although Brucella abortus, one of the disease's etiologic agents, possesses
cytokine-stimulatory properties, the mechanism by which this bacterium
triggers a proinflammatory response is not known. We examined the mechanism
whereby heat-killed B. abortus (HKBA), as well as its LPS, induces
production of inflammatory cytokines in monocytes/macrophages. Polymyxin B,
a specific inhibitor of LPS activity, did not inhibit the production of
TNF-alpha- and IL-6-induced HKBA in the human monocytic cell line THP-1.
HKBA induced the production of these cytokines in peritoneal macrophages of
both C3H/HeJ and C3H/HeN mice, whereas B. abortus LPS only stimulated cells
from C3H/HeN mice. Anti-TLR2 Ab, but not anti-TLR4 Ab, blocked HKBA-mediated
TNF-alpha and IL-6 production in THP-1 cells. Because bacterial
lipoproteins, a TLR2 ligand, have potent inherent stimulatory properties, we
investigated the capacity of two B. abortus lipoproteins, outer membrane
protein 19 (Omp19) and Omp16, to elicit a proinflammatory response. Lipidated (L)-Omp16 and L-Omp19, but not their unlipidated forms, induced
the secretion of TNF-alpha, IL-6, IL-10, and IL-12 in a time- and
dose-dependent fashion. Preincubation of THP-1 cells with anti-TLR2 Ab
blocked L-Omp19-mediated TNF-alpha and IL-6 production. Together, these
results entail a mechanism whereby B. abortus can stimulate cells from the
innate immune system and induce cytokine-mediated inflammation in
brucellosis. We submit that LPS is not the cause of inflammation in
brucellosis; rather, lipoproteins of this organism trigger the production of
proinflammatory cytokines, and TLR2 is involved in this process.
http://www.ncbi.nlm.nih.gov/pubmed/15383598
http://www.jimmunol.org/cgi/content/full/173/7/4635
2004;
Lipid-associated membrane proteins of Mycoplasma fermentans and M. penetrans activate human immunodeficiency virus long-terminal repeats through
Toll-like receptors.
Mycoplasmas are known to enhance human
immunodeficiency virus (HIV) replication, and mycoplasma-derived lipid
extracts have been reported to activate nuclear factor-kappaB (NF-kappaB)
through Toll-like receptors (TLRs). In this study, we examined the
involvement of TLRs in the activation of HIV long-terminal repeats (LTR) by
mycoplasma and their active components responsible for the TLR activation.
Lipid-associated membrane proteins (LAMPs) from two species of mycoplasma
(Mycoplasma fermentans and M. penetrans) that are associated with acquired
immune-deficiency syndrome (AIDS), were found to activate HIV LTRs in a
human monocytic cell line, THP-1. NF-kappaB deletion from the LTR
resulted in inhibition of the activation. The LTR activation by M.
fermentans LAMPs was inhibited by a dominant negative (DN) construct of TLR1
and TLR6, whereas HIV LTR activation by M. penetrans LAMPs was inhibited by
DN TLR1, but not by DN TLR6. These results indicate that the activation
of HIV LTRs by M. fermentans and M. penetrans LAMPs is dependent on NF-kappaB,
and that the activation of HIV LTR by M. fermentans LAMPs is mediated
through TLR1, TLR2 and TLR6. In contrast, the LTR activation by M.
penetrans LAMPs is carried out through TLR1 and TLR2, but not TLR6.
Subsequently, the active component of M. penetrans and M. fermentans LAMPs
was purified by reverse-phase high-performance liquid chromatography (HPLC).
Interestingly, the purified lipoprotein of M. penetrans LAMPs (LPMp) was
able to activate NF-kappaB through TLR1 and TLR2. On the other hand, the
activation of NF-kappaB by purified lipoprotein of M. fermentans LAMPs (LPMf)
was mediated through TLR2 and TLR6, but not TLR1.
http://www.ncbi.nlm.nih.gov/pubmed/15312143
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1782549/?tool=pubmed
2004, Jun;
Synthetic lipopeptide adjuvants and Toll-like
receptor 2--structure-activity relationships.
Bacterial lipoproteins and their synthetic analogues (sLP) are strong immune
modulators of the early host responses after infection. Synthetic lipopeptides
are strong adjuvants for the adaptive immune system. Lipoproteins and
lipopeptides induce signalling in immune cells through Toll-like
receptor-TLR2/TLR1 heterodimers
[IMAGE:
http://www.pdbj.org/eprots/index_en.cgi?PDB%3A2Z7X]. By screening a combinatorial lipohexapeptide
amide collection in an in vitro IL-8 induction assay, we systematically
evaluated the potential of 19 proteinogenic amino acids in the peptide moiety of
Pam3Cys-lipopeptides to interact with TLR2. New Pam3Cys-lipopeptides with high
activity were obtained. Different fatty acids were introduced to investigate the
influence of the acyl moiety. Lipopeptides with modifications in the core
structure of the unusual amino acid S-glycerylcysteine were synthesized and
tested for IL-8 induction via TLR2.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
2004;
Tolerance induced by the lipopeptide
Pam3Cys is due to ablation of
IL-1R-associated kinase-1.
Stimulation of the human monocytic cell line Mono Mac 6 with the synthetic
lipopeptide
(S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys(4)-OH,
trihydrochloride (Pam(3)Cys) at 10 microg/ml induces a rapid expression of the
TNF gene in a TLR2-dependent fashion. Preculture of the cells with Pam(3)Cys at
1 microg/ml leads to a reduced response after subsequent stimulation with
Pam(3)Cys at 10 microg/ml, indicating that the cells have become tolerant to
Pam(3)Cys. The CD14 and TLR2 expression is not decreased on the surface of the
tolerant cells, but rather up-regulated. Analysis of the NF-kappaB binding in
Pam(3)Cys-tolerant cells shows a failure to mobilize NF-kappaB-p50p65
heterodimers, while NF-kappaB-p50p50 homodimers remain unchanged.
Pam(3)Cys-tolerant cells showed neither IkappaBalpha-Ser(32) phosphorylation nor
IkappaBalpha degradation but MyD88 protein was unaltered. However, IRAK-1
protein was absent in Pam(3)Cys-induced tolerance, while IRAK-1 mRNA was still
detectable at 30% compared with untreated cells. In contrast, in LPS-tolerized
cells, p50p50 homodimers were induced, IRAK-1 protein level was only partially
decreased, and p50p65 mobilization remained intact. It is concluded that in Mono
Mac 6 monocytic cells, inhibition of IRAK-1 expression at the mRNA and protein
levels is
the main TLR-2-dependent mechanism responsible for Pam(3)Cys-induced
tolerance, but not for TLR-4-dependent LPS-induced tolerance.
http://www.ncbi.nlm.nih.gov/pubmed/15294992
2004;
Blood; Mycoplasma fermentans infection promotes immortalization of human
peripheral blood mononuclear cells in culture.
Chronic infection or colonization by mycoplasma(s)
could gradually and significantly alter many biologic properties of mammalian
host cells in culture, including induction of malignant transformation. We
examined effects of Mycoplasma fermentans infection on the continuing survival
and immortality of human peripheral blood mononuclear cells (PBMCs) from healthy
blood donors. Without specific supplemental growth factors, human PBMCs normally
die rapidly, with few cells other than macrophages/monocytes surviving after 2
weeks in cultures. Only occasional Epstein-Barr virus (EBV)-positive B
lymphocytes would continue to proliferate and undergo spontaneous
immortalization. Our present study revealed that infection of human PBMCs in
culture with the incognitus and PG18 strains of M fermentans, but surprisingly
not with some other strains tested in parallel, markedly enhanced the rate of
EBV-positive B lymphocytes to undergo immortalization (74% vs 17%). Compared
with spontaneously immortalized PBMCs, the PBMCs immortalized in cultures
infected with the mycoplasmas often had prominent karyotype changes with
chromosomal loss, gain, or translocations. Furthermore, many of these
immortalized B lymphocytes were found to be monoclonal in nature. The in vitro
findings would be of relevance to lymphoproliferative disorders that occurred in
patients with immune suppression. The mycoplasma-mediated promotional effect in
cell immortalization and its potential clinical implications warrant further
study.
http://www.ncbi.nlm.nih.gov/pubmed/15331449
SEE ALL RELATED:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=15331449
2004;
Lipid-associated membrane proteins of Mycoplasma fermentans and M. penetrans activate human immunodeficiency virus
long-terminal repeats through Toll-like receptors.
Mycoplasmas are known to
enhance human immunodeficiency virus
(HIV) replication, and
mycoplasma-derived lipid extracts have
been reported to activate nuclear
factor-kappaB (NF-kappaB) through
Toll-like receptors (TLRs).
In this study, we examined the
involvement of TLRs in the activation of
HIV long-terminal repeats (LTR) by mycoplasma and their active components responsible for the TLR
activation. Lipid-associated membrane proteins (LAMPs) from two species of
mycoplasma (Mycoplasma fermentans and M. penetrans) that are associated with
acquired immune-deficiency syndrome (AIDS), were found to activate HIV LTRs in a
human monocytic cell line, THP-1. NF-kappaB deletion from the LTR resulted in
inhibition of the activation. The LTR activation by M. fermentans LAMPs was
inhibited by a dominant negative (DN) construct of TLR1 and TLR6, whereas HIV
LTR activation by M. penetrans LAMPs was inhibited by DN TLR1, but not by DN
TLR6. These results indicate that the activation of HIV LTRs by M. fermentans
and M. penetrans LAMPs is dependent on NF-kappaB, and that
the activation of HIV LTR by M. fermentans LAMPs is mediated through TLR1, TLR2 and TLR6. In contrast,
the LTR activation by M. penetrans LAMPs is carried out through TLR1 and TLR2,
but not TLR6. Subsequently, the active component of M. penetrans and M.
fermentans LAMPs was purified by reverse-phase high-performance liquid
chromatography (HPLC). Interestingly, the purified lipoprotein of M. penetrans
LAMPs (LPMp) was able to activate NF-kappaB through TLR1 and TLR2. On the other
hand, the activation of NF-kappaB by purified lipoprotein of M. fermentans LAMPs
(LPMf) was mediated through TLR2 and TLR6, but not TLR1.
http://www.ncbi.nlm.nih.gov/pubmed/15312143
2004;
Tolerance induced by the lipopeptide Pam3Cys is
due to ablation of IL-1R-associated kinase-1.
Stimulation of the human monocytic cell line Mono Mac 6 with the synthetic
lipopeptide
(S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys(4)-OH,
trihydrochloride (Pam(3)Cys) at 10 microg/ml induces a rapid expression of the
TNF gene in a TLR2-dependent fashion. Preculture of the cells with Pam(3)Cys at
1 microg/ml leads to a reduced response after subsequent stimulation with
Pam(3)Cys at 10 microg/ml, indicating that
the cells have become tolerant to
Pam(3)Cys. The CD14 and TLR2 expression is not decreased on the surface of the
tolerant cells, but rather up-regulated. Analysis of the NF-kappaB binding in
Pam(3)Cys-tolerant cells shows a failure to mobilize NF-kappaB-p50p65 heterodimers, while NF-kappaB-p50p50 homodimers remain unchanged.
Pam(3)Cys-tolerant cells showed neither IkappaBalpha-Ser(32) phosphorylation nor
IkappaBalpha degradation but MyD88 protein was unaltered. However, IRAK-1
protein was absent in Pam(3)Cys-induced tolerance, while IRAK-1 mRNA was still
detectable at 30% compared with untreated cells. In contrast, in LPS-tolerized
cells, p50p50 homodimers were induced, IRAK-1 protein level was only partially
decreased, and p50p65 mobilization remained intact. It is concluded that in Mono
Mac 6 monocytic cells, inhibition of IRAK-1 expression at the mRNA and protein
levels is the main TLR-2-dependent mechanism
responsible for Pam(3)Cys-induced
tolerance, but not for TLR-4-dependent LPS-induced tolerance.
http://www.ncbi.nlm.nih.gov/pubmed/15294992
2004, Aug; Lipopolysaccharide binding
protein binds to triacylated and
diacylated lipopeptides and mediates
innate immune responses.
LPS binding protein (LBP) is an acute-phase protein synthesized predominantly in the liver of the mammalian host. It was first described to bind LPS of Gram-negative bacteria and transfer it via a CD14-enhanced mechanism to a receptor complex including TLR-4 and MD-2, initiating a signal transduction cascade leading to the release of proinflammatory cytokines. In recent studies, we found that LBP also mediates cytokine induction caused by compounds derived from Gram-positive bacteria, including lipoteichoic acid and peptidoglycan fragments. Lipoproteins and lipopeptides have repeatedly been shown to act as potent cytokine inducers, interacting with TLR-2, in synergy with TLR-1 or -6. In this study, we show that these compounds also interact with LBP and CD14. We used triacylated lipopeptides, corresponding to lipoproteins of Borrelia burgdorferi, mycobacteria, and Escherichia coli, as well as diacylated lipopeptides, corresponding to, e.g., 2-kDa macrophage activating lipopeptide of Mycoplasma spp. Activation of Chinese hamster ovary cells transfected with TLR-2 by both lipopeptides was enhanced by cotransfection of CD14. Responsiveness of human mononuclear cells to these compounds was greatly enhanced in the presence of human LBP. Binding of lipopeptides to LBP as well as competitive inhibition of this interaction by LPS was demonstrated in a microplate assay. Furthermore, we were able to show that LBP transfers lipopeptides to CD14 on human monocytes using FACS analysis. These results support that LBP is a pattern recognition receptor transferring a variety of bacterial ligands including the two major types of lipopeptides to CD14 present in different receptor complexes.
http://www.ncbi.nlm.nih.gov/pubmed/15294986
http://www.jimmunol.org/content/173/4/2683.long
2005;
Presence of Epstein-Barr virus and human herpesvirus 6B DNA in
multiple sclerosis patients: associations with disease activity.
OBJECTIVE:
To assess the presence of
Epstein-Barr virus (EBV) and human herpesvirus 6B (HHV-6B) DNA in saliva and
plasma from multiple sclerosis (MS) patients enrolled in a randomized,
double-blind, placebo-controlled valacyclovir treatment study.
METHODS: DNA was prepared
following ultracentrifugation of saliva and plasma. EBV and HHV-6B DNAs were
determined by real-time polymerase chain reaction.
RESULTS: EBV and HHV-6B DNAs
were detected in 41% and 65% of saliva samples, respectively. In patients
treated with valacyclovir, the percentage of saliva samples with EBV was
significantly reduced (9%; P = 0.000017), whereas the frequency of HHV-6B
positive samples was unchanged (57%; P = 0.38). Longitudinal studies
demonstrated a time-dependent reduction in the frequency of saliva samples
containing EBV following valacyclovir treatment. In contrast, plasma
contained EBV and HHV-6B DNAs in 17% and 25% of the samples, respectively,
and these numbers were not significantly reduced following valacylovir
treatment (13% and 16%, respectively), nor were they different from those of
healthy controls (6% and 39%, respectively). Patients with high disease
activity had a significantly higher frequency of EBV (P = 0.018) and HHV-6B
(P = 0.023) positive samples than did patients with low disease activity.
The presence of EBV and HHV-6B was strongly correlated in plasma (P <
0.00000001), but not in saliva (P = 0.41).
CONCLUSION: MS patients express
EBV and HHV-6B in both saliva and plasma, but only the expression of EBV in
saliva is significantly reduced following valacyclovir treatment. Although
EBV and HHV-6B DNAs can be detected in plasma from healthy individuals, the
co-expression of both these viruses in MS patients is highly significant and
further associated with clinical activity. The observations of viral DNA in
plasma are consistent with an underlying immunologic defect in MS.
http://www.ncbi.nlm.nih.gov/pubmed/16281923
2005, Jun;
Role of TLR in B cell development: signaling
through TLR4 promotes B cell maturation and is inhibited by TLR2.
The role of TLR4 in mature B cell activation is well characterized. However,
little is known about TLR4 role in B cell development. Here, we analyzed the
effects of TLR4 and TLR2 agonists on B cell development using an in vitro model
of B cell maturation. Highly purified B220(+)IgM(-) B cell precursors from
normal C57BL/6 mouse were cultured for 72 h, and B cell maturation in the
presence of the TLR agonists was evaluated by expression of IgM, IgD, CD23, and
AA4. The addition of LPS or lipid A resulted in a marked increase in the
percentage of CD23(+) B cells, while Pam3Cys had no effect alone, but inhibited
the increase of CD23(+) B cell population induced by lipid A or LPS. The
TLR4-induced expression of CD23 is not accompanied by full activation of the
lymphocyte, as suggested by the absence of activation Ag CD69. Experiments with
TLR2-knockout mice confirmed that the inhibitory effects of Pam3Cys depend on
the expression of TLR2. We studied the effects of TLR-agonists on early steps of
B cell differentiation by analyzing IL-7 responsiveness and phenotype of early B
cell precursors: we found that both lipid A and Pam3Cys impaired IL-7-dependent
proliferation; however, while lipid A up-regulates B220 surface marker,
consistent with a more mature phenotype of the IgM(-) precursors, Pam3Cys keeps
the precursors on a more immature stage. Taken together, our results suggest
that TLR4 signaling favors B lymphocyte maturation, whereas TLR2 arrests/retards
that process, ascribing new roles for TLRs in B cell physiology.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
http://www.jimmunol.org/cgi/content/full/174/11/6639
You should be getting a clearer picture
by now that OspA/Pam3Cys could not have
been a vaccine
and this is the reason IDSociety, Yale,
and now the Tribune continue to torture
us
2005, Latov; Antibodies against OspA
epitopes of Borrelia burgdorferi
cross-react with neural tissue.
Neurological sequela of chronic Lyme disease include encephalopathy, myelopathy and peripheral neuropathy. These have generally been attributed to either persistent infection or pathogen-induced autoimmunity. In this study, we investigated the presence of cross-reactive human neural epitopes that share amino acid sequences with Borrelia burgdorferi OspA protein. Sequence similarity analysis was carried out by searching known cDNA sequences from brain tissue. The cDNA database search yielded three sequences that were identical to sequences in OspA. Corresponding peptides were synthesized and antibodies were generated against them in rabbits. Antibodies against two of the homologous OspA peptides were found to react with neurons in human brain, spinal cord and dorsal root ganglia by immunohistochemistry.
http://www.ncbi.nlm.nih.gov/pubmed/15652419
2005;
[Guillain-Barré Syndrome and its association with
infectious factors.]
Guillain-Barré Syndrome (GBS) is an acute polyneuropathy often triggered by
inflammatory and probably autoimmune mechanisms. Development of GBS is in 2/3 of
cases preceded by acute infection, typically with gastrointestinal or
respiratory symptoms. Infectious agents related to
GBS include cytomegalovirus,
Epstein-Barr virus, Campylobacter jejuni, Mycoplasma pneumoniae and Haemophilus
influenzae. Molecular mimicry seems to be responsible for GBS development after
infection,
through the synthesis of autoantibodies against myelin gangliosides.
Autoimmune reactions develop only in a small fraction of all exposed
individuals, depending on still unresolved factors. Different infections lead to
forms of GBS differing in spectrum of autoantibodies and in frequency with which
different clinical symptoms appear. This may be of some significance for early
prognosis and in future possibly for choosing therapeutic options. An increased
risk of GBS may be also related to vaccination, but with presently used vaccines
this increase remains below one case of GBS per one million doses.
http://www.ncbi.nlm.nih.gov/pubmed/15981163
See autoantibodies
to gangliosides as a common feature of
neuroborreliosis - yet now denied as an
outcome of LYMErix or Lyme disease.
1995, Benach; Experimental immunization with Borrelia burgdorferi induces development of antibodies to gangliosides.
Benach, http://www.ncbi.nlm.nih.gov/pubmed/7558329
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC173580/?tool=pubmed
This was among the data given to Richard Blumenthal and the US Attorney's office which was stolen off of Mr. Blumenthal's desk by Jessica Gauvin (Oct 4, 2003 dataset) It has never been explained how Gauvin stole Blumenthal's mail, since we know this to be a federal office, yet HERE IT IS.
1995, Benach; Borrelia burgdorferi shows specificity of binding to glycosphingolipids.
http://www.ncbi.nlm.nih.gov/pubmed/7622201
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC173381/?tool=pubmed
2006;
Interactions between mycoplasma lipid-associated
membrane proteins and the host cells.
Mycoplamas are a group of wall-less prokaryotes widely distributed in nature,
some of which are pathogenic for humans and animals. There are many lipoproteins
anchored on the outer face of the plasma membrane, called lipid-associated
membrane proteins (LAMPs). LAMPs are highly antigenic and could undergo phase
and size variation, and are recognized by the innate immune system through
Toll-like receptors (TLR) 2 and 6. LAMPs can modulate the immune system, and
could induce immune cells apoptosis or death. In addition, they may associate
with malignant transformation of host cells and are also considered to be
cofactors in the progression of AIDS.
http://www.ncbi.nlm.nih.gov/pubmed/16615163
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1462930/?tool=pubmed
2005;
Inhibition of neutrophil apoptosis by
TLR agonists in whole blood: involvement
of the phosphoinositide 3-kinase/Akt and
NF-kappaB signaling pathways, leading to
increased levels of Mcl-1, A1, and
phosphorylated Bad.
Using flow cytometry, we investigated the effect of TLR agonists on human polymorphonuclear neutrophil (PMN) apoptosis in whole blood. LPS (TLR4), peptidoglycan (TLR2), R-848 (TLR7/8), and CpG-DNA (TLR9) were equally effective at delaying spontaneous apoptosis of PMN, while PamCSK4 (TLR1/2), macrophage-activating lipopeptide-2 (TLR2/6), flagellin (TLR5), and loxoribine (TLR7) were less effective or inactive. TLR agonists found to delay apoptosis also extended the functional life span of PMN. Analysis of signaling pathways revealed that the antiapoptotic effect of TLR agonists required NF-kappaB and PI3K activation. Furthermore, analysis of intact cells by flow cytometry showed that TLR agonists delaying PMN apoptosis increased phosphorylation of Akt, a major target of PI3K. This effect was associated with a PI3K-dependent increase in heat shock protein 27 phosphorylation, which has been reported to play a key role in PMN survival. Finally, the TLR-induced delay in PMN apoptosis was associated with increased levels of Mcl-1 and A1, which are antiapoptotic members of the Bcl-2 family. These effects were reversed by PI3K and NF-kappaB inhibitors, respectively. TLR activation also led to PI3K-dependent phosphorylation of the proapoptotic protein Bad. Taken together, our results strongly suggest a role of NF-kappaB and PI3K in TLR-induced PMN survival, leading to modulation of Bcl-2 family molecules.
http://www.ncbi.nlm.nih.gov/pubmed/15749901
2006, Jan, CDC says Chronic Fatigue
Syndrome is due to cell-cycle changes
etc post Epstein-Barr infection:
Preliminary evidence of mitochondrial dysfunction associated with post-infective fatigue after acute infection with Epstein Barr virus.
ABSTRACT: Acute infectious diseases are typically accompanied by non-specific symptoms including fever, malaise, irritability and somnolence that usually resolve on recovery. However, in some individuals these symptoms persist in what is commonly termed post-infective fatigue. The objective of this pilot study was to determine the gene expression correlates of post-infective fatigue following acute Epstein Barr virus (EBV) infection.
METHODS: We followed 5 people with acute mononucleosis who developed post-infective fatigue of more than 6 months duration and 5 HLA-matched control subjects who recovered within 3 months. Subjects had peripheral blood mononuclear cell (PBMC) samples collected at varying time points including at diagnosis, then every 2 weeks for 3 months, then every 3 months for a year. Total RNA was extracted from the PBMC samples and hybridized to microarrays spotted with 3,800 oligonucleotides.
RESULTS: Those who developed post-infective fatigue had gene expression profiles indicative of an altered host response during acute mononucleosis compared to those who recovered uneventfully. Several genes including ISG20 (interferon stimulated gene), DNAJB2 (DnaJ [Hsp40] homolog and CD99), CDK8 (cyclin-dependent kinase 8), E2F2 (E2F transcription factor 2), CDK8 (cyclin-dependent kinase 8), and ACTN2 (actinin, alpha 2), known to be regulated during EBV infection, were differentially expressed in post-infective fatigue cases. Several of the differentially expressed genes affect mitochondrial functions including fatty acid metabolism and the cell cycle.
CONCLUSION: These preliminary data provide insights into alterations in gene transcripts associated with the varied clinical outcomes from acute infectious mononucleosis.
http://www.ncbi.nlm.nih.gov/pubmed/16448567
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1373655/?tool=pubmed
CDC References this article from 1948:
http://www.ncbi.nlm.nih.gov/pubmed/18873530 "Chronic infectious mononucleosis."
2006;
Toll-like receptor 2 controls expansion and
function of regulatory T cells.
Tregs play a central role
in the suppression of immune reactions and prevention of autoimmune responses
harmful to the host. During acute infection, however, Tregs might hinder
effector T cell activity directed toward the elimination of the pathogenic
challenge. Pathogen recognition receptors from the TLR family expressed by
innate immune cells are crucial for the generation of effective immunity. We
have recently shown the CD4CD25 Treg subset in TLR2 mice to be significantly
reduced in number compared with WT littermate control mice, indicating a link
between Tregs and TLR2.
Here, we report that the TLR2 ligand Pam3Cys, but not
LPS (TLR4) or CpG (TLR9), directly acts on purified Tregs in a MyD88-dependent
fashion. Moreover, when combined with TCR stimulation, TLR2 triggering augmented
Treg proliferation in vitro and in vivo and resulted in a temporal loss of the
suppressive Treg phenotype in vitro by directly affecting Tregs. Importantly, WT Tregs adoptively transferred into TLR2 mice were neutralized by systemic
administration of TLR2 ligand during the acute phase of a Candida albicans
infection, resulting in a 100-fold reduced C. albicans outgrowth. This
demonstrates that in vivo TLR2 also controls the function of Tregs and
establishes a direct link between TLRs and the control of immune responses
through Tregs.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
See the
latest from Allen Steere on tregs.
2006, Jun;
Weismuller;
TLR1- and
TLR6-independent recognition of
bacterial lipopeptides.
Bacterial cell walls contain lipoproteins/peptides, which are strong modulators of the innate immune system. Triacylated lipopeptides are assumed to be recognized by TLR2/TLR1-, whereas diacylated lipopeptides use TLR2/TLR6 heteromers for signaling. Following our initial discovery of TLR6-independent diacylated lipopeptides, we could now characterize di- and triacylated lipopeptides (e.g. Pam(2)C-SK(4), Pam(3)C-GNNDESNISFKEK), which have stimulatory activity in TLR1- and in TLR6-deficient mice. Furthermore, for the first time, we present triacylated lipopeptides with short length ester-bound fatty acids (like PamOct(2)C-SSNASK(4)), which induce no response in TLR1-deficient cells. No differences in the phosphorylation of MAP kinases by lipopeptide analogs having different TLR2-coreceptor usage were observed. Blocking experiments indicated that different TLR2 heteromers recognize their specific lipopeptide ligands independently from each other. In summary, a triacylation pattern is necessary but not sufficient to render a lipopeptide TLR1-dependent, and a diacylation pattern is necessary but not sufficient to render a lipopeptide TLR6-dependent. Contrary to the current model, distinct lipopeptides are recognized by TLR2 in a TLR1- and TLR6-independent manner.
http://www.ncbi.nlm.nih.gov/pubmed/16455646
http://www.jbc.org/content/281/14/9049.long
The S-(2,3-dihydroxypropyl)-cysteine may be acylated with a third amide-linked fatty acid, as is the case for the LP from the cell wall of Escherichia coli (13) and its synthetic lipohexapeptide analog Pam3C-SK4 (14). Interestingly, human and murine cells show a species-specific difference in the TLR2-dependent recognition of distinct LP structures (15), which seems to be linked to the chain length of the O-acylated fatty acids (16).
2006, Sep;
Bilateral facial palsy: Epstein-Barr virus, not Lyme disease.
Bilateral facial palsy is frequently linked
with lyme disease. We report a patient with bilateral facial palsy due to
Epstein-Barr virus infection but with Borrelia burgdorferi IgM in serum
caused by polyclonal B-lymphocyte stimulation.
http://www.ncbi.nlm.nih.gov/pubmed/16930373
2006, Oct: Human cellular protein
VRK2 interacts specifically with
Epstein-Barr virus BHRF1, a homologue of
Bcl-2, and enhances cell survival.
BHRF1, an early gene product of Epstein-Barr virus (EBV), is structurally and functionally homologous to Bcl-2, a cellular anti-apoptotic protein. BHRF1 has been shown to protect cells from apoptosis induced by numerous external stimuli. Nasopharyngeal carcinoma is an epithelial cancer associated closely with EBV infection. Specific proteins that might interact with and modulate the BHRF1 anti-apoptotic activity in normal epithelial cells are of interest. Therefore, a cDNA library derived from normal human foreskin keratinocytes was screened by the yeast two-hybrid system and a cellular gene encoding human vaccinia virus B1R kinase-related kinase 2 (VRK2) was isolated. Interaction between the cellular VRK2 and viral BHRF1 proteins was further demonstrated by glutathione S-transferase pull-down assays, confocal laser-scanning microscopy and co-immunoprecipitation. Analyses of VRK2-deletion mutants revealed that a 108 aa fragment at the C terminus was important for VRK2 to interact with BHRF1. For BHRF1, aa 1-18 and 89-142 were crucial in interacting with VRK2 and these two regions are counterparts of Bcl-2 homology domains 4 and 1. Overexpressed VRK2 alone showed a modest effect in anti-apoptosis and appeared to enhance cell survival in the presence of BHRF1. However, this enhancement was not observed when VRK2 was co-expressed with Bcl-2. The results indicate that human VRK2 interacts specifically with EBV BHRF1 and that the interaction is involved in protecting cells from apoptosis.
http://www.ncbi.nlm.nih.gov/pubmed/16963744
http://vir.sgmjournals.org/cgi/content/full/87/10/2869
http://www.ncbi.nlm.nih.gov/pubmed?term=anti-apoptosis[All%20Fields]%20AND%20epstein-barr[All%20Fields]&cmd=DetailsSearch: (10 references):
Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) forms complexes with a cellular anti-
apoptosis protein Bcl-2 or its EBV counterpart BHRF1 through HS1-associated protein X-1.
STAT3 activation contributes directly to Epstein-Barr virus-mediated invasiveness of nasopharyngeal cancer cells in vitro.
2006, Philipp;
Interleukin-10
anti-inflammatory response to Borrelia burgdorferi, the agent of Lyme disease: a
possible role for suppressors of cytokine signaling 1 and 3.
It has been established that interleukin-10
(IL-10) inhibits inflammatory cytokines produced by macrophages in response
to Borrelia burgdorferi or its lipoproteins. The mechanism by which IL-10
exerts this anti-inflammatory effect is still unknown. Recent findings
indicate that suppressors of cytokine signaling (SOCS) proteins are induced
by cytokines and Toll-like receptor (TLR)-mediated stimuli, and in turn they
can down-regulate cytokine and TLR signaling in macrophages. Because it is
known that SOCS are induced by IL-10 and that B. burgdorferi and its
lipoproteins most likely interact via TLR2 or the heterodimers TLR2/1 and/or
TLR2/6, we hypothesized that SOCS are induced by IL-10 and B. burgdorferi
and its lipoproteins in macrophages and that SOCS may mediate the inhibition
by IL-10 of concomitantly elicited cytokines. We report here that mouse J774
macrophages incubated with IL-10 and added B. burgdorferi spirochetes
(freeze-thawed, live, or sonicated) or lipidated outer surface protein A
(L-OspA) augmented their SOCS1/SOCS3 mRNA and protein expression, with SOCS3
being more abundant. Pam(3)Cys, a synthetic lipopeptide, also induced
SOCS1/SOCS3 expression under these conditions, but unlipidated OspA was
ineffective. Neither endogenous IL-10 nor the translation inhibitor cycloheximide blocked SOCS1/SOCS3 induction by B. burgdorferi and its
lipoproteins, indicating that the expression of other genes is not required.
This temporally correlated with the IL-10-mediated inhibition of the
inflammatory cytokines IL-1beta, IL-6, IL-12p40, IL-18, and tumor necrosis
factor alpha. Our data are evidence to suggest that expression of SOCS is
part of the mechanism of IL-10-mediated inhibition of inflammatory cytokines
elicited by B. burgdorferi and its lipoproteins.
http://www.ncbi.nlm.nih.gov/pubmed/16988256
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1594918/?tool=pubmed
2006;
Mycobacterium tuberculosis subverts
innate immunity to evade specific
effectors.
The macrophage is the niche of the intracellular pathogen Mycobacterium tuberculosis. Induction of macrophage apoptosis by CD4(+) or CD8(+) T cells is accompanied by reduced bacterial counts, potentially defining a host defense mechanism. We have already established that M. tuberculosis-infected primary human macrophages have a reduced susceptibility to Fas ligand (FasL)-induced apoptosis. To study the mechanisms by which M. tuberculosis prevents apoptotic signaling, we have generated a cell culture system based on PMA- and IFN-gamma-differentiated THP-1 cells recapitulating the properties of primary macrophages. In these cells, nucleotide-binding oligomerization domain 2 or TLR2 agonists and mycobacterial infection protected macrophages from apoptosis and resulted in NF-kappaB nuclear translocation associated with up-regulation of the antiapoptotic cellular FLIP. Transduction of a receptor-interacting protein-2 dominant-negative construct showed that nucleotide-binding oligomerization domain 2 is not involved in protection in the mycobacterial infection system. In contrast, both a dominant-negative construct of the MyD88 adaptor and an NF-kappaB inhibitor abrogated the protection against FasL-mediated apoptosis, showing the implication of TLR2-mediated activation of NF-kappaB in apoptosis protection in infected macrophages. The apoptosis resistance of infected macrophages might be considered as an immune escape mechanism, whereby M. tuberculosis subverts innate immunity signaling to protect its host cell against FasL(+)-specific cytotoxic lymphocytes.
http://www.ncbi.nlm.nih.gov/pubmed/17056554
http://www.jimmunol.org/cgi/content/full/177/9/6245
2006, NINDS' Roland
Martin on OspA as the cause of the MS
outcome of Lyme:
Borrelia burgdorferi Induces TLR1 and
TLR2 in human microglia and peripheral
blood monocytes but differentially
regulates HLA-class II expression.
"The
spirochete Borrelia burgdorferi is the
agent of Lyme disease, which causes
central nervous system manifestations in
up to 20% of patients. We investigated
the response of human brain microglial
cells, glial progenitors, neurons,
astrocytes, as well as peripheral blood
monocytes to stimulation with B.
burgdorferi. We used oligoarrays to
detect changes in the expression of
genes important for shaping adaptive and
innate immune responses. We found that
stimulation with B. burgdorferi lysate
increased the expression of Toll-like
receptors (TLRs) 1 and 2 in all cell
types except neurons. However, despite
similarities in global gene profiles of
monocytes and microglia, only microglial
cells responded to the stimulation with
a robust increase in HLA-DR, HLA-DQ, and
also coexpressed CD11-c, a dendritic
cell marker. In contrast, a large number
of HLA-related molecules were repressed
at both the RNA and the protein levels
in stimulated monocytes, whereas
secretion of IL-10 and TNF-alpha was
strongly induced. These results show
that signaling through TLR1/2 in
response to B. burgdorferi can elicit
opposite immunoregulatory effects in
blood and in brain immune cells, which
could play a role in the different
susceptibility of these compartments to
infection."
http://www.ncbi.nlm.nih.gov/pubmed/16783164
2007; MICE ARE
TERRIBLE MODELS OF "LYME DISEASE":
Differential
expression of Toll-like receptor 2
(TLR2) and responses to TLR2 ligands
between human and murine vascular
endothelial cells.
J Endotoxin Res. 2007;13(5):281-96.
Shuang
Chen,
Wong MH,
Schulte DJ,
Arditi M,
Michelsen KS.
Division of Pediatric Infectious Diseases, Burns and Allen Research Institute, Cedars-Sinai Medical Center, David Geffen School of Medicine at UCLA, Los Angeles, California 94008, USA.
Toll-like receptors (TLRs) initiate and maintain host defenses and inflammation, and directly contribute to diseases such as atherosclerosis. It is not completely understood in what cell types proatherogenic TLR-induced signaling arises and, particularly, there is uncertainty regarding the potential functional role of TLR2 in endothelial cells (ECs). We determined TLR2 and TLR4 gene expression in four different human and two different murine primary ECs using gene array analysis, RT-PCR, and flow cytometry and confirmed these data by functional studies by stimulating ECs with the corresponding TLR ligands. TLR4 was expressed in all human and murine ECs and these cells responded to stimulation with LPS. Faint expression of TLR2 was observed in human ECs, whereas murine ECs express considerable amounts of TLR2 mRNA. Human ECs failed to respond to TLR2 ligands while murine ECs responded to TLR2 ligands. Furthermore, in murine ECs, TLR2 was located on the cell surface while in human ECs, TLR2 was sequestered in intracellular compartments. After IFN-gamma or IL-1beta stimulation, TLR2 translocated to the cell surface of human ECs. In conclusion, TLR2 is expressed intracellularly in human ECs and, therefore, TLR2 ligands are inaccessible to the receptor. Murine ECs express membrane TLR2 and respond to TLR2 ligands, but human ECs normally will not respond unless they are first primed with inflammatory stimulation, which appears to trigger translocation of TLR2 to the cell surface. http://www.ncbi.nlm.nih.gov/pubmed/17986487
But we knew that
years ago, when it was reported that
"mouse brains are not
a good support system for spirochetes:"
http://www.actionlyme.org/HOW_RICO_WILL_BE_CHARGED.htm
Felsenfeld
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=441260&blobtype=pdf
2007;
Epstein-Barr virus induces MCP-1 secretion by
human monocytes via TLR2.
Epstein-Barr virus (EBV) is a
gammaherpesvirus infecting the majority of
the human adult population in the world. TLR2, a member of the Toll-like
receptor (TLR) family, has been implicated in the immune responses to different
viruses including members of the herpesvirus family, such as human
cytomegalovirus, herpes simplex virus type 1, and varicella-zoster virus. In
this report, we demonstrate that infectious and UV-inactivated EBV virions lead
to the activation of NF-kappaB through TLR2 using HEK293 cells cotransfected
with TLR2-expressing vector along with NF-kappaB-Luc reporter plasmid. NF-kappaB
activation in HEK293-TLR2 cells (HEK293 cells transfected with TLR2) by EBV was
not enhanced by the presence of CD14. The effect of EBV was abrogated by
pretreating HEK293-TLR2 cells with blocking anti-TLR2 antibodies or by
preincubating viral particles with neutralizing anti-EBV antibodies 72A1. In
addition, EBV infection of primary human monocytes induced the release of MCP-1
(monocyte chemotactic protein 1), and the use of small interfering RNA targeting
TLR2 significantly reduced such a chemokine response to EBV. Taken together,
these results indicate that TLR2 may be an important pattern recognition
receptor in the immune response directed against EBV infection.
http://www.ncbi.nlm.nih.gov/pubmed/17522215
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1951286/?tool=pubmed
2007;
Increased prevalence of
varicella zoster virus DNA in cerebrospinal fluid from patients with multiple
sclerosis.
In order to investigate the possible
involvement of viruses in Multiple Sclerosis (MS), the study evaluated the
presence of viral genomic sequences in cerebrospinal fluid (CSF), as markers
of viral replication within the central nervous system (CNS). A total of 85
CSF samples were collected from 38 MS patients, 28 patients with other
neurological diseases and 19 subjects without neurological diseases. Using
nested-PCR, the investigation focused on the presence of human herpes virus
DNA, including herpes simplex virus 1 (HSV-1) and 2 (HSV-2), the
Epstein-Barr virus (EBV), varicella zoster virus (VZV), human
cytomegalovirus (HCMV), human herpes virus 6 (HHV-6) and JC virus (JCV). All
the CSF samples from the individuals without neurological diseases were
negative for viral DNA. Genomic sequences of HSV-1, HCMV, EBV, HHV6, and JCV
were found in patients with MS and other neurological diseases without
significant differences between the two groups. VZV DNA was detected more
frequently (P < 0.05) in the MS group (31.6%), particularly among the
relapsing-remitting MS patients (43.5%), compared with patients with other
neurological diseases (10.7%). In addition, the results indicated that JCV
and HHV-6 were replicating actively in the CNS of a small, but significant
number of patients with MS and other neurological diseases. Most
importantly, the study revealed a high frequency of VZV DNA in the CSF of
patients with MS, suggesting a possible role of this virus in the
pathogenesis of MS.
http://www.ncbi.nlm.nih.gov/pubmed/17177306
2007, Aug; A common human TLR1
polymorphism regulates the innate immune
response to lipopeptides.
Toll-like receptors (TLR) are critical mediators of the immune response to pathogens and human polymorphisms in this gene family regulate inflammatory pathways and are associated with susceptibility to infection. Lipopeptides are present in a wide variety of microbes and stimulate immune responses through TLR1/2 or TLR2/6 heterodimers. It is not currently known whether polymorphisms in TLR1 regulate the innate immune response. We stimulated human whole blood with triacylated lipopeptide, a ligand for TLR1/2 heterodimers, and found substantial inter-individual variation in the immune response. We sequenced the coding region of TLR1 and found a non-synonymous polymorphism, I602S (base pair T1805G), that regulated signalling. In comparison to TLR1_602S, the 602I variant mediated substantially greater basal and lipopeptide-induced NF-kappaB signalling in transfected HEK293 cells. These signalling differences among TLR1 variants were also found with stimulation by extracts of Mycobacterium tuberculosis. Furthermore, individuals with the 602II genotype produced substantially more IL-6 than those with the 602SS variant in a lipopeptide-stimulated whole-blood cytokine assay. Together, these observations demonstrate that variation in the inflammatory response to bacterial lipopeptides is regulated by a common TLR1 transmembrane domain polymorphism that could potentially impact the innate immune response and clinical susceptibility to a wide spectrum of pathogens.
http://www.ncbi.nlm.nih.gov/pubmed/17595679
CIted By:
http://www.ncbi.nlm.nih.gov/pubmed?db=pubmed&cmd=link&linkname=pubmed_pubmed_citedin&uid=17595679
We're looking for the link between Steere's HLA outcomes and normal people and wondering if the effect is upstream, here in TLR2 or TLR1 changes..
2007;
Mycoplasmal membrane protein p37 promotes
malignant changes in mammalian cells.
Evidence of Mycoplasma hyorhinis infection in human gastric cancer tissues has
been found in previous work. In this study, we demonstrate that the expression
of p37, a membrane lipoprotein of M. hyorhinis, in mammalian cells induces
antisenescence, enhances clonogenicity in soft agar, and co-operates with human
epidermal growth factor receptor-related 2 to inhibit cell adhesion. Conversely,
truncated p37 protein, with the first 28 amino acids deleted from its N
terminal, promotes cell senescence. Taken together, our findings suggest that
p37 promotes malignant changes in mammalian cells. With the identification of
this molecular component, which is responsible for mycoplasma
malignancy-promoting activity, it is possible that a better understanding of the
relationship between M. hyorhinis infection and human gastric cancers will lead
to novel diagnostics and therapeutics.
http://www.ncbi.nlm.nih.gov/pubmed/17496976
2007, Nov;
MALT lymphoma : recent advances in aetiology
and molecular genetics.
Mucosa-associated lymphoid tissue (MALT) lymphoma is a common low grade B-cell
lymphoma arising from a background of chronic inflammatory disease at a number
of mucosal sites. Those originating in the stomach are causatively linked to
Helicobacter pylori infection and eradication of the bacterium with antibiotics
leads to long-term complete regression of the lymphoma in aproximately 70% of
cases. Now, there is further evidence of linking
Campylobacter jejuni, Borrelia burgdorferi
and Chlamydia psittaci
infection with immunoproliferative small intestine disease, MALT lymphoma of the
skin and ocular adnexa respectively. t(11;18)/API2-MALT1, t(1;14)/IGH-BCL10,
t(14;18)/IGH-MALT1 and t(3;14)/IGH-FOXP1 occur at considerably variable
incidences in MALT lymphomas of different sites. The first three chromosome
translocations are specifically associated with the MALT lymphoma entity and the
oncogenic products of these translocations have been shown to target a common
molecular pathway, i.e. the nuclear factor-kappaB pathway. Here, I review the
recent advances in our understanding of the association of microbial pathogens
with MALT lymphoma of various sites and the molecular genetics underlying the
lymphoma development.
http://www.ncbi.nlm.nih.gov/pubmed/18040143
http://www.jstage.jst.go.jp/article/jslrt/47/2/31/_pdf
2007;
The inhibitory effect of Mycoplasma fermentans on tumour necrosis factor (TNF)-alpha-induced apoptosis resides in the membrane
lipoproteins.
Mycoplasma have been shown to be involved in the alteration of several
eukaryotic cell functions, such as cytokine production, gene expression and
more. We have previously reported that infection of human myelomonocytic U937
cell line with live Mycoplasma fermentans (M. fermentans) inhibited tumour
necrosis factor (TNF-alpha)-induced apoptosis. Mycoplasmal membrane lipoproteins
are considered to be the most potent initiators of inflammatory reactions in
mycoplasmal infections. The aim of this study was to clarify whether the
inhibitory effect on TNFalpha-induced apoptosis is exerted by M. fermentans
lipoproteins (LPMf). A significant reduction in TNFalpha-induced apoptosis was
demonstrated by stimulation of U937 cells with M. fermentans total proteins,
LPMf or MALP-2 (M. fermentans synthetic lipopeptide), but not with M. fermentans
hydrophilic protein preparation (AqMf). To investigate the mechanism of M.
fermentans antiapoptotic effect, the reduction of mitochondrial transmembrane
potential (delta psi m) was measured. M. fermentans total proteins LPMf and
MALP-2, but not AqMf, inhibited the reduction of delta psi m. In addition, M.
fermentans total proteins LPMf and MALP-2, but not AqMf, downregulated the
formation of active caspase-8. NF-kappaB was transactivated in cells treated
with M. fermentans lipoproteins, and was essential for host cell survival, but
not for the inhibition of TNFalpha-induced apoptosis by LPMf. Our results
suggest that the inhibitory effect exerted by M. fermentans on TNFalpha-induced
apoptosis in U937 cells is due to the membrane lipoproteins of these bacteria. http://www.ncbi.nlm.nih.gov/pubmed/16889623
SEE ALL
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=16889623
2008; Jan,
Brucella abortus inhibits major histocompatibility complex
class II expression and antigen processing through interleukin-6 secretion via
Toll-like receptor 2.
See Don Scott and the Stealth "Crystal," also:
http://www.youtube.com/KMDickson#p/a/u/0/Umwk_sTJmWk
The strategies that allow Brucella abortus
to survive inside macrophages for prolonged periods and to avoid the
immunological surveillance of major histocompatibility complex class II
(MHC-II)-restricted gamma interferon (IFN-gamma)-producing CD4+ T
lymphocytes are poorly understood. We report here that infection of THP-1
cells with B. abortus inhibited expression of MHC-II molecules and antigen
(Ag) processing. Heat-killed B. abortus (HKBA) also induced both these
phenomena, indicating the independence of bacterial viability and
involvement of a structural component of the bacterium. Accordingly,
outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein,
inhibited both MHC-II expression and Ag processing to the same extent as
HKBA. Moreover, a synthetic lipohexapeptide that mimics the structure of the
protein lipid moiety also inhibited MHC-II expression, indicating that any
Brucella lipoprotein could down-modulate MHC-II expression and Ag
processing. Inhibition of MHC-II expression and Ag processing by either
HKBA or lipidated Omp19 (L-Omp19) depended on Toll-like receptor 2 and was
mediated by interleukin-6. HKBA or L-Omp19 also inhibited MHC-II expression
and Ag processing of human monocytes. In addition, exposure to the synthetic
lipohexapeptide inhibited Ag-specific T-cell proliferation and IFN-gamma
production of peripheral blood mononuclear cells from Brucella-infected
patients. Together, these results indicate that there is a mechanism by
which B. abortus may prevent recognition by T cells to evade host immunity
and establish a chronic infection.
http://www.ncbi.nlm.nih.gov/pubmed/17984211
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2223644/?tool=pubmed
2008, March, the
Pam3Cys and
Biomarkers Chapters of
Cryme Disease published.
2008, May,
Regulation and dysregulation of Epstein-Barr
virus latency: implications for the development of autoimmune diseases.
Epstein-Barr virus (EBV) is
a human herpesvirus hiding in a latent form in memory B cells in the majority of
the world population. Although, primary EBV infection is asymptomatic or causes
a self-limiting disease, infectious mononucleosis, the virus is associated with
a wide variety of neoplasms developing in immunosuppressed or immunodeficient
individuals, but also in patients with an apparently intact immune system. In
memory B cells, tumor cells, and lymphoblastoid cell lines (LCLs, transformed by
EBV in vitro) the expression of the viral genes is highly restricted. There is
no virus production (lytic viral replication associated with the expression of
all viral genes) in tight latency. The expression of latent viral oncogenes and
RNAs is under a strict epigenetic control via DNA methylation and histone
modifications that results either in a complete silencing of the EBV genome in
memory B cells, or in a cell-type dependent usage of latent promoters in tumor
cells, germinal center B cells, and LCLs. Both the latent and lytic EBV proteins
are potent immunogens and elicit vigorous B- and T-cell responses.
In
immunosuppressed and immunodeficient patients, or in individuals with a
functional defect of EBV-specific T cells, lytic EBV replication is regularly
activated and an increased viral load can be detected in the blood. Enhanced
lytic replication results in new infection events and EBV-associated
transformation events, and seems to be a risk factor both for malignant
transformation and the development of autoimmune diseases. One may speculate
that an increased load or altered presentation of a limited set of lytic or
latent EBV proteins that cross-react with cellular antigens triggers and
perpetuates the pathogenic processes that result in multiple sclerosis, systemic
lupus erythematosus (SLE), and rheumatoid arthritis.
In addition, in SLE
patients EBV may cause defects of B-cell tolerance checkpoints because latent
membrane protein 1, an EBV-encoded viral oncoprotein can induce BAFF, a B-cell
activating factor that rescues self-reactive B cells and induces a lupus-like
autoimmune disease in
transgenic mice.
http://www.ncbi.nlm.nih.gov/pubmed/18432410
2008;
Tolerance induced via TLR2 and TLR4 in human dendritic cells: role of IRAK-1.
BACKGROUND:
While dendritic cells (DCs) can induce tolerance
in T cells, little is known about tolerance induction in DCs themselves. We have
analysed tolerance induced in human in-vitro generated DCs by repeated
stimulation with ligands for TLR4 and TLR2.
RESULTS:
DCs stimulated with the TLR4 ligand LPS did show
a rapid and pronounced expression of TNF mRNA and protein. When DCs were
pre-cultured for 2 days with 5 ng LPS/ml then the subsequent response to
stimulation with a high dose of LPS (500 ng/ml) was strongly reduced for both
TNF mRNA and protein. At the promoter level there was a reduced transactivation
by the -1173 bp TNF promoter and by a construct with a tetrameric NF-kappaB
motif. Within the signalling cascade leading to NF-kappaB activation we found an
ablation of the IRAK-1 adaptor protein in LPS-tolerant DCs. Pre-culture of DCs
with the TLR2 ligand Pam3Cys also led to tolerance with respect to TNF gene
expression and IRAK-1 protein was ablated in such tolerant cells as well, while
IRAK-4 protein levels were unchanged.
CONCLUSION:
These data show that TLR-ligands can render DCs
tolerant with respect to TNF gene expression by a mechanism that likely involves
blockade of signal transduction at the level of IRAK-1.
http://www.ncbi.nlm.nih.gov/pubmed/19025640
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2628880/?tool=pubmed
2008, Aug;
Preconditioning with a TLR2 specific ligand
increases resistance to cerebral ischemia/reperfusion injury.
The brain's resistance to
ischemic injury can be transiently augmented by prior exposure to a sub-lethal
stress stimulus, i.e. preconditioning. It has been reported that Toll-like
receptors (TLRs) are involved in the preconditioning-induced protective effect
against ischemic brain injury. In this study, we investigated the effect of
preconditioning with a TLR2 specific ligand,
Pam3CSK4,
on focal cerebral ischemia/reperfusion (I/R) injury in mice. Pam3CSK4 was
administered systemically 24 h before the mice were subjected to focal cerebral
ischemia (1 h) followed by reperfusion.
Cerebral infarct size was
determined, blood brain barrier (BBB) permeability was evaluated, and expression
of tight-junction proteins were examined after focal cerebral I/R. Results
showed that pre-treatment with Pam3CSK significantly reduced brain infarct size
(1.9+/-0.5% vs 9.4+/-2.2%) compared with the untreated I/R group.
Pam3CSK4
pre-treatment also significantly reduced acute mortality (4.3% vs 24.2%),
preserved neurological function (8.22+/-0.64 vs 3.91+/-0.57), and attenuated
brain edema (84.61+/-0.08% vs 85.29+/-0.09%) after cerebral I/R. In addition,
Pam3CSK4 pre-treatment preserved BBB function as evidenced by decreased leakage
of serum albumin (0.528+/-0.026 vs 0.771+/-0.059) and Evans Blue (9.23+/-0.72 microg/mg vs 12.56+/-0.65 microg/mg) into brain tissue. Pam3CSK4 pre-treatment
also attenuated the loss of the tight junction protein occludin in response to
brain I/R injury. These results suggest that TLR2 is a new target of ischemic
preconditioning in the brain and preconditioning with a TLR2 specific ligand
will protect the brain from I/R injury.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
Or Lithium will do the same thing.
2008;
Human immunodeficiency virus infection alters
tumor necrosis factor alpha production via Toll-like receptor-dependent pathways
in alveolar macrophages and U1 cells.
Human immunodeficiency virus
(HIV)-positive persons are predisposed to pulmonary infections, even after
receiving effective highly active antiretroviral therapy. The reasons for this
are unclear but may involve changes in innate immune function. HIV type 1
infection of macrophages impairs effector functions, including cytokine
production. We observed decreased constitutive tumor necrosis factor alpha
(TNF-alpha) concentrations and increased soluble tumor necrosis factor receptor
type II (sTNFRII) in bronchoalveolar lavage fluid samples from HIV-positive
subjects compared to healthy controls. Moreover, net proinflammatory TNF-alpha
activity, as measured by the TNF-alpha/sTNFRII ratio, decreased as HIV-related
disease progressed, as manifested by decreasing CD4 cell count and increasing
HIV RNA (viral load). Since TNF-alpha is an important component of the innate
immune system and is produced upon activation of Toll-like receptor (TLR)
pathways, we hypothesized that the mechanism associated with deficient TNF-alpha
production in the lung involved altered TLR expression or a deficit in the TLR
signaling cascade. We found decreased Toll-like receptor 1 (TLR1) and TLR4
surface expression in HIV-infected U1 monocytic cells compared to the uninfected
parental U937 cell line and decreased TLR message in alveolar macrophages (AMs)
from HIV-positive subjects. In addition, stimulation with TLR1/2 ligand
(Pam(3)Cys) or TLR4 ligand (lipopolysaccharide) resulted in decreased
intracellular phosphorylated extracellular signal-regulated kinase and
subsequent decreased transcription and expression of TNF-alpha in U1 cells
compared to U937 cells. AMs from HIV-positive subjects also showed decreased
TNF-alpha production in response to these TLR2 and TLR4 ligands. We postulate
that HIV infection alters expression of TLRs with subsequent changes in mitogen-activated protein kinase signaling and cytokine production that
ultimately leads to deficiencies of innate immune responses that predispose
HIV-positive subjects to infection.
http://www.ncbi.nlm.nih.gov/pubmed/18524817
2008; Feb;
Induction of EBV-latent membrane protein
1-specific MHC class II-restricted T-cell responses against natural killer
lymphoma cells.
EBV-encoded latent membrane protein 1
(LMP1) has oncogenic potential and is expressed in many EBV-associated
malignancies. Although LMP1 is regarded as a potential tumor-associated
antigen for immunotherapy and several LMP1-specific MHC class I–restricted
CTL epitopes have been reported, little is known regarding MHC class
II–restricted CD4 helper T-lymphocyte (HTL) epitopes for LMP1. The goal of
the present studies was to determine whether MHC class II–restricted CD4
T-cell responses could be induced against the LMP1 antigen and to evaluate
the antitumor effect of these responses. We have combined the use of a
predictive MHC class II binding peptide algorithm with in vitro
vaccination of CD4 T cells using candidate peptides to identify naturally
processed epitopes derived from LMP1 that elicit immune responses against
EBV-expressing tumor cells. Peptide LMP1159-175 was effective in
inducing HTL responses that were restricted by HLA-DR9, HLA-DR53, or
HLA-DR15, indicating that this peptide behaves as a promiscuous T-cell
epitope. Moreover, LMP1159-175–reactive HTL clones directly
recognized EBV lymphoblastoid B cells, EBV-infected natural killer
(NK)/T-lymphoma cells and naturally processed antigen in the form of LMP1+
tumor cell lysates presented by autologous dendritic cells. Because the
newly identified epitope LMP1159-175 overlaps with an
HLA-A2–restricted CTL epitope (LMP1159-167), this peptide might
have the ability to induce simultaneous CTL and HTL responses against LMP1.
Overall, our data should be relevant for the design and optimization of
T-cell epitope–based immunotherapy against various EBV-associated
malignancies, including NK/T cell lymphomas. [Cancer Res 2008;68(3):901–8]
LMP1 is required for the EBV-mediated
transformation of B lymphocytes and has attracted significant interest
because, thus far, it is the only EBV latency gene capable of transforming
rodent fibroblasts in vitro, which are tumorgenic in nude mice (
9). Furthermore, it has been reported that B-cell lymphomas can
spontaneously arise in LMP1 transgenic mice without the need of additional
EBV genes (
10). In addition, LMP1 interacts with several signaling proteins of the
tumor necrosis factor receptor family (
11) and Janus kinase 3 (
12), resulting in nuclear factor-κB (
13) and activator protein 1 (
14) induction and activation. Moreover, LMP1 protects against apoptosis
by increasing bcl-2 activity (
15). In view of the above, LMP1 is considered as the main EBV-derived
oncoprotein and has become an ideal target for T cell–based immunotherapy
against EBV-induced malignancies.
http://www.ncbi.nlm.nih.gov/pubmed/18245493
SEE ALL:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=18245493
p66 and
OspA are an ABC-transporter complex and
p66 is a porin
2008, Dec; The Triacylated ATP
Binding Cluster Transporter
Substrate-binding Lipoprotein of
Staphylococcus aureus Functions as a
Native Ligand for Toll-like Receptor 2.
Some synthetic lipopeptides, in addition
to native lipoproteins derived from both
Gram-negative bacteria and mycoplasmas,
are known to activate TLR2 (Toll-like
receptor 2). However, the native
lipoproteins inherent to Gram-positive
bacteria, which function as TLR2
ligands, have not been characterized.
Here, we have purified a native
lipoprotein to homogeneity from
Staphylococcus aureus to study as a
native TLR2 ligand. The purified 33-kDa
lipoprotein was capable of stimulating
TLR2 and was identified as a triacylated
SitC lipoprotein, which belongs to a
family of ATP binding cluster (ABC)
transporter substrate-binding proteins.
Analyses of the SitC-mediated production
of cytokine using mouse peritoneal
macrophages revealed that the SitC
protein (3 nm) induced the production of
tumor necrosis factor-alpha and
interleukin-6. Moreover, analysis of
knock-out mice showed that SitC required
TLR2 and MyD88, but not TLR1 or TLR6,
for the induction of cytokines. In
addition to the S. aureus SitC
lipoprotein, we purified two other
native ABC transporter substrate-binding
lipoproteins from Bacillus subtilis and
Micrococcus luteus, which were both
shown to stimulate TLR2. These results
demonstrate that S. aureus SitC
lipoprotein is triacylated and that the
ABC transporter substrate-binding
lipoproteins of Gram-positive bacteria
function as native ligands for TLR2
http://www.ncbi.nlm.nih.gov/pubmed/19139093
please read:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2659198/?tool=pubmed
2009,
March; Mapping of a microbial protein
domain involved in binding and
activation of the TLR2/TLR1 heterodimer.
The pentameric B subunit of type IIb Escherichia coli enterotoxin (LT-IIb-B(5)), a doughnut-shaped oligomeric protein from enterotoxigenic E. coli, activates the TLR2/TLR1 heterodimer (TLR2/1). We investigated the molecular basis of the LT-IIb-B(5) interaction with TLR2/1 to define the structure-function relationship of LT-IIb-B(5) and, moreover, to gain an insight into how TLR2/1 recognizes large, nonacylated protein ligands that cannot fit within its lipid-binding pockets, as previously shown for the Pam(3)CysSerLys(4) (Pam(3)CSK(4)) lipopeptide. We first identified four critical residues in the upper region of the LT-IIb-B(5) pore. Corresponding point mutants (M69E, A70D, L73E, S74D) were defective in binding TLR2 or TLR1 and could not activate APCs, despite retaining full ganglioside-binding capacity. Point mutations in the TLR2/1 dimer interface, as determined in the crystallographic structure of the TLR2/1-Pam(3)CSK(4) complex, resulted in diminished activation by both Pam(3)CSK(4) and LT-IIb-B(5). Docking analysis of the LT-IIb-B(5) interaction with this apparently predominant activation conformation of TLR2/1 revealed that LT-IIb-B(5) might primarily contact the convex surface of the TLR2 central domain. Although the TLR1/LT-IIb-B(5) interface is relatively smaller, the leucine-rich repeat motifs 9-12 in the central domain of TLR1 were found to be critical for cooperative TLR2-induced cell activation by LT-IIb-B(5). Moreover, the putative LT-IIb-B(5) binding site overlaps partially with that of Pam(3)CSK(4); consistent with this, Pam(3)CSK(4) suppressed TLR2 binding of LT-IIb-B(5), albeit not as potently as self-competitive inhibition. We identified the upper pore region of LT-IIb-B(5) as a TLR2/1 interactive domain, which contacts the heterodimeric receptor at a site that is distinct from, although it overlaps with, that of Pam(3)CSK(4).
http://www.ncbi.nlm.nih.gov/pubmed/19234193
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2648068/?tool=pubmed
2009, April;
A TLR2
ligand suppresses inflammation by
modulation of chemokine receptors and
redirection of leukocyte migration.
Toll-like receptors orchestrate rapid local protective innate-immune responses to invading pathogens and optimize leukocyte priming of subsequent adaptive responses. Paradoxically, systemic excess of the TLR2 ligand, bacterial lipoprotein (BLP), suppresses peripheral inflammatory responses. Here, we demonstrate that this phenomenon is regulated via the TLR2-dependent, cell-autonomous down-regulation of inflammatory chemokine receptor expression on a variety of leukocyte subsets. Remarkably, BLP mediated no effect on constitutive chemokine receptor expression. By tracking adoptively transferred wild-type and TLR2(-/-) leukocytes in vivo, we observed that BLP mediated chemokine receptor switching directed leukocytes away from inflamed sites toward secondary lymphoid organs. These data highlight a novel role for TLR ligands, such as BLP, in regulating leukocyte retention and migration away from innate immune lesions via discrete constitutive and inflammatory chemokine receptor regulation.
http://www.ncbi.nlm.nih.gov/pubmed/19202130
If you can understand that this is a SCIENTIFIC FACT, then you can understand that the qualification of LYMErix must have been a crime:
"The proinflammatory nature of TLR signaling is logical in the context of mounting inflammatory responses to localized microbial infection. However, several studies have reported, paradoxically, that systemic exposure to some TLR ligands (including bacterial lipoprotein [BLP] and lipopolysaccharide [LPS], ***ligands for TLRs 2 and 4, respectively) is anti-inflammatory and can suppress*** subsequent leukocyte accumulation in response to a variety of locally applied inflammatory stimuli.5–8"
http://bloodjournal.hematologylibrary.org/cgi/content/full/113/18/4224
2009;
A TLR2 ligand suppresses allergic inflammatory
reactions by acting directly on mast cells.
BACKGROUND:
Although much attention has been
focused on the anti-allergic effects of probiotics, their mode of action is not
fully understood. Mast cells, which play a central role in inducing allergic
inflammation, are potential targets of probiotics given the recent discovery
that they express Toll-like receptors (TLRs), the pattern recognition receptors
for microbial components. In this study, we examined whether allergic reactions
of mast cells are modulated by stimulation through TLR2.
METHODS:
The effects on mast cells of the synthetic TLR2
ligand Pam3CSK4 and Bifidobacterium pseudocatenulatum JCM 7041 were evaluated in
vitro. Furthermore, the effects of Pam3CSK4 on mast cell-induced increase in
vascular permeability in vivo were investigated by employing mast cell-deficient
W/W(v) mice into which IgE-sensitized mouse bone marrow-derived mast cells were
transferred.
RESULTS:
Pam3CSK4 and Bifidobacterium pseudocatenulatum
JCM 7041 suppressed degranulation of IgE-sensitized mast cells upon antigen
stimulation in vitro. Pam3CSK4 also suppressed leukotriene C(4) production
triggered by engagement of the high-affinity IgE receptor, FcepsilonRI.
Intracellular Ca(2+) mobilization and phosphorylation of Erk were suppressed by
pretreatment with Pam3CSK4, suggesting that the TLR2 ligand suppresses
activation of mast cells by interrupting FcepsilonRI-mediated intracellular
signaling. Pam3CSK4 treatment of bone marrow-derived mast cells reduced the
increase in vascular permeability in recipient W/W(v) mice upon intravenous
injection of antigen; the decrease was by about half, in a TLR-dependent manner.
CONCLUSION:
Collectively, these results demonstrate that the
FcepsilonRI-mediated inflammatory responses of mast cells are suppressed by
stimulation through TLR2, suggesting that probiotics exert potential
anti-allergic effects, at least in part, through direct effects on mast cells.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
2009, July;
Soluble TLR2 reduces inflammation
without compromising bacterial clearance
by
disrupting TLR2 triggering.
TLR overactivation may lead to end organ damage and serious acute and chronic inflammatory conditions. TLR responses must therefore be tightly regulated to control disease outcomes. We show in this study the ability of the soluble form of TLR2 (sTLR2) to regulate proinflammatory responses, and demonstrate the mechanisms underlying sTLR2 regulatory capacity. Cells overexpressing sTLR2, or stimulated in the presence of the sTLR2 protein, are hyporesponsive to TLR2 ligands. Regulation was TLR2 specific, and affected NF-kappaB activation, phagocytosis, and superoxide production. Natural sTLR2-depleted serum rendered leukocytes hypersensitive to TLR2-mediated stimulation. Mice administered sTLR2 together with Gram-positive bacteria-derived components showed lower peritoneal levels of the neutrophil (PMN) chemoattractant, keratinocyte-derived chemokine; lower PMN numbers; and a reduction in late apoptotic PMN. Mononuclear cell recruitment remained unaffected, and endogenous peritoneal sTLR2 levels increased. Notably, the capacity of sTLR2 to modulate acute inflammatory parameters did not compromise the ability of mice to clear live Gram-positive bacteria-induced infection. Mechanistically, sTLR2 interfered with TLR2 mobilization to lipid rafts for signaling, acted as a decoy microbial receptor, and disrupted the interaction of TLR2 with its coreceptor, CD14, by associating with CD14. These findings establish sTLR2 as a regulator of TLR2-mediated inflammatory responses, capable of blunting immune responses without abrogating microbial recognition and may inform the design of novel therapeutics against acute and chronic inflammatory conditions.
http://www.ncbi.nlm.nih.gov/pubmed/19542461
2009;
Down-Regulation of MHC Class II Expression through
Inhibition of CIITA Transcription by Lytic Transactivator Zta during
Epstein-Barr Virus Reactivation1
It
has been reported that the lytic cycle of EBV correlates with the
diminution of cell surface MHC class I molecules, and down-regulation
of surface MHC class I expression is maintained throughout the lytic
cycle of EBV, which could conceivably have a significant effect on Ag
presentation (13).
In EBV-negative B cells, Zta completely inhibits the up-regulation of
surface MHC class I expression induced, and also directly inhibits
the constitutive activation of NF-B
(13,
14). In addition, it has
recently been shown that the expression
of the IFN-receptor gene is down-regulated at both the mRNA and protein levels
following introduction of an adenovirus vector expressing Zta (15).
It
has also been proposed that MHC class II molecules and the Ag-processing pathway may be targets of EBV for escaping CD4+
T
lymphocyte immunosurveillance. The studies with human CMV (HCMV) and
mouse CMV reveal that CMV blocks IFN--inducible
MHC class II transcription (16,
17). By inhibiting Jak1 expression
and disrupting IFN--stimulated
signaling pathway, HCMV interferes with MHC class II transcription
and CIITA activation. Other strategies for the down-modulation MHC
class II surface expression by HCMV are promoting the
proteasome-mediated degradation of DR-and DM-molecules by glycoprotein US2 (18,
19). In addition,
US3 protein of HCMV competes with Ii for
binding to MHC class II molecules, interfering with Ag presentation (19,
20). Only
recently have analogous immune evasion strategies been
described for EBV. It has been demonstrated that the EBV gp42 protein
binds to HLA class II molecules at their various stages of maturation
and impairs TCR-mediated activation of Ag-specific Th cells
in
an HLA class II-dependent manner (21).
A truncated soluble EBV gp42 protein, generated by proteolytic
cleavage of full-length gp42 in the endoplasmic reticulum during the
EBV lytic infection, can mediate Ag-specific evasion from Th
cell recognition by blocking the TCR-HLA class II-peptide association
(22).
It has also been reported that EBV lytic infection is associated with
the reduction of MHC class II expression (13).
However, the mechanism underlying modulation of MHC class II
expression by EBV has not yet been established.
MHC class II is exquisitely controlled mainly at the level of
transcription initiation by a highly conserved regulatory module.
A
very tight regulation of MHC class II expression is crucial for the
control of the immune response. CIITA is the major regulator of MHC
class II gene expression and functions as a non-DNA-binding transcriptional coactivator by interacting with almost all of
the
transcription regulatory proteins, forming a stable enhanceosome bound to the SXY regulatory module of MHC class II promoters
and
driving transcription of MHC class II genes (23,
24).
The present study aims to explore how MHC class II expression
was
down-regulated in EBV lytic cycle. We first observed that the
expression of MHC class II was reduced in Raji cells induced by 12-O-tetradecanoylphorbol-13-acetate
(TPA) and sodium butyrate (NaB) concomitantly with a significant
decrease of CIITA expression. Computational analysis revealed two
potential ZREs within the CIITA promoter. Experimental studies
demonstrated that Zta inhibited the promoter activity and
transcription of CIITA. The binding of Zta to the promoter of CIITA
was confirmed both in vitro and in vivo. Furthermore, the expression
of MHC class II and CIITA was restored by small-interfering
(si)RNA-directed knockdown of Zta. Our data indicate that
Zta-mediated suppression of CIITA
transcription is a distinct
mechanism of down-regulation of MHC class II expression during EBV
reactivation.
http://www.jimmunol.org/cgi/content/full/182/4/1799
http://www.ncbi.nlm.nih.gov/pubmed/19201831
2009; Infectious agents and lymphoma development: molecular and clinical
aspects.
This review is focused on the role of
infectious agents in the development of some lymphoma entities. Associations
involving bacterial infections mostly regard marginal zone B-cell lymphomas
of mucosa-associated lymphoid tissue (MALT)-type. Some paradigmatic examples
of these associations include the Helicobacter pylori-related gastric MALT
lymphoma and the more recently reported links between Chlamydophila psittaci
and ocular adnexal MALT lymphomas and Borrelia burgdorferi and cutaneous
MALT lymphomas. The well-documented association between Epstein-Barr virus
infection and related lymphoproliferative disorders are analysed as an
example of lymphotropic virus with tumourigenic activity. Molecular,
biological and clinical features as well as therapeutic implications of
these associations are analysed and future perspectives in this field are
discussed.
http://www.ncbi.nlm.nih.gov/pubmed/19298458
2009, Sep,
Persistent exposure to Mycoplasma induces
malignant transformation of human prostate cells.
Recent
epidemiologic, genetic, and molecular studies suggest infection and inflammation
initiate certain cancers, including those of the prostate. The American Cancer
Society, estimates that approximately 20% of all worldwide cancers are caused by
infection. Mycoplasma, a genus of bacteria that lack a cell wall, are among the
few prokaryotes that can grow in close relationship with mammalian cells, often
without any apparent pathology, for extended periods of time. In this study, the
capacity of Mycoplasma genitalium, a prevalent sexually transmitted infection,
and Mycoplasma hyorhinis, a mycoplasma found at unusually high frequency among
patients with AIDS, to induce a malignant phenotype in benign human prostate
cells (BPH-1) was evaluated using a series of in vitro and in vivo assays. After
19 weeks of culture, infected BPH-1 cells achieved anchorage-independent growth
and increased migration and invasion. Malignant transformation of infected BPH-1
cells was confirmed by the formation of xenograft tumors in athymic mice.
Associated with these changes was an increase in karyotypic entropy, evident by
the accumulation of chromosomal aberrations and polysomy. This is the first
report describing the capacity of M. genitalium or M. hyorhinis infection to
lead to the malignant transformation of benign human epithelial cells and may
serve as a model to further study the relationship between prostatitis and
prostatic carcinogenesis.
http://www.ncbi.nlm.nih.gov/pubmed/19721714
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2730529/?tool=pubmed
2009;
Allen Steere's *LIES* about seronegative Lyme to Volkman:
Allen Steere feels he has to keep lying
about seronegative Lyme because that way he can deny OspA's Great Imitator/Immunosupression
outcomes. "Lyme Disease" is Allen Steere's falsified testing version
of it. That's why people have to start referring to this as Relapsing
Fever. "Lyme Disease" is an imaginary thing that happened in Europe,
conducted by Allen Steere, alone, which was meant to falsify LYMErix'
outcome and over which Willy Burgdorfer called Allen Steere both retarded
and a liar:
http://underourskin.com/blog/?p=191
Dr. Burgdorfer:
"The controversy in Lyme disease research is a shameful affair. And I say
that because the whole thing is politically tainted. Money goes to people
who have, for the past 30 years, produced the same thing—nothing.
Serology has to be started from scratch with people who don’t know
beforehand the results of their research."
Yale:
"Sex Cures All Diseases" and "Scientific discovery could interfere
with the intended propaganda or marketing of LYMErix"
2009;
Anti-apoptotic genes in the survival of monocytic cells during infection.
Macrophages are cells of the immune system that protect organisms against invading pathogens by fulfilling critical roles in innate and adaptive immunity and inflammation. They originate from circulating monocytes and show a high degree of heterogeneity, which reflects the specialization of function given by different anatomical locations. Differentiation of monocytes towards a macrophage phenotype is also accompanied by an increase of resistance against various apoptotic stimuli, a required characteristic that allows macrophages to accomplish their function in a stressful environment.Apoptosis, a form of programmed cell death, is a tightly regulated process, needed to maintain homeostasis by balancing proliferation with cellular demise. Caspases, a family of cysteine proteases that are highly conserved in multicellular organisms, function as central regulators of apoptosis. FLIP (FLICE-inhibitory protein), anti-apoptotic members of the Bcl2 family and inhibitors of apoptosis (IAP) are the main three groups of anti-apoptotic genes that counteract caspase activation through both the extrinsic and intrinsic apoptotic pathways. Modulation of the apoptotic machinery during viral and bacterial infections, as well as in various malignancies, is a well established mechanism that promotes the survival of affected cells. The involvement of anti-apoptotic genes in the survival of monocytes/macrophages, either physiological or pathological, will be described in this review. How viral and bacterial infections that target cells of the monocytic lineage affect the expression of anti-apoptotic genes is important in understanding the pathological mechanisms that lead to manifested disease. The latest therapeutic approaches that target anti-apoptotic genes will also be discussed.
http://www.ncbi.nlm.nih.gov/pubmed/20119528
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2729995/?tool=pubmed
M.tb also exploits TLRs to induce anti-apoptotic genes that enhance cell survival and promote bacterial persistence [109]. Exploiting TLRs is not a mechanism unique to M.tb. As mentioned earlier, we have shown that TLR3, TLR4 and TLR9, when stimulated by their ligands PolyI:C, LPS and CpG DNA, respectively, protected monocytic cells from HIV-Vpr induced apoptosis by induction of NFκB and anti-apoptotic cIAP genes (unpublished data).
Stimulation of TLR2, found in abundance at sites of M.tb infection, by components of M.tb cell wall, has been shown to protect human macrophages against apoptosis. THP1-derived macrophages when stimulated with 19kDa mycobacterial lipoprotein or mannosylated LAM were shown to induce resistance to apoptosis via activation of NFκB and subsequent induction of anti apoptotic cFLIP which inhibits death receptor-mediated apoptosis [25, 109].
2009;
Decreased T cell reactivity to
Epstein-Barr virus infected
lymphoblastoid cell lines in multiple
sclerosis.
OBJECTIVE: To investigate T cell and antibody immunity to Epstein-Barr virus (EBV) in multiple sclerosis (MS).
METHODS: Immunoglobulin G (IgG) immunity to EBV nuclear antigen 1 (EBNA1) and viral capsid antigen was measured by enzyme linked immunosorbent assays, and T cell immunity was assessed using enzyme linked immunospot assays to measure the frequency of peripheral blood mononuclear cells (PBMC) producing interferon gamma in response to autologous EBV infected B cell lymphoblastoid cell lines (LCL) in 34 EBV seropositive healthy subjects and 34 EBV seropositive patients with MS who had not received immunomodulatory therapy in the previous 3 months.
RESULTS: Patients with MS had increased levels of anti-EBNA1 IgG but a decreased frequency of LCL specific T cells compared with healthy subjects. Using purified populations of CD4(+) T cells and CD8(+) T cells, we showed that the LCL specific response resides predominantly in the CD8(+) population, with a frequency 5-7-fold higher than in the CD4(+) population. The decreased CD8(+) T cell response to LCL in MS was not caused by decreased HLA class I expression by LCL, and LCL from MS patients could be killed normally by HLA matched EBV specific cytotoxic CD8(+) T cell clones from healthy subjects. Furthermore, the decreased CD8(+) T cell immunity to EBV was not due to a primary defect in the function of CD8(+) T cells because EBV specific cytotoxic CD8(+) T cell lines could be generated normally from the PBMC of patients with MS.
CONCLUSION: This quantitative deficiency in CD8(+) T cell immunity to EBV might be responsible for the accumulation of EBV infected B cells in the brains of patients with MS.
http://www.ncbi.nlm.nih.gov/pubmed/19015225
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2663364/?tool=pubmed
2010, Jan; Chinese investigate
how to reverse/prevent the immune damage
of the
Newest Great Imitator,
LYMErix, which results in all those
horrible outcomes of Lyme and LYMErix
[activated
latent viral infections, tolerance to
mycoplasma (ALS);
Gulf War
Illness, Chronic Fatigue/
Activated, , Chronic
Active Mono (Epstein-Barr), Cancer,
MS, FibroFemino-NotEnoughSex-itis
Lupus, etc.]:
[The influence of over expression
of interleukin-1 receptor-associated
kinase 1 on bacterial
lipoprotein-induced tolerance.]
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue.
2010 Jan;22(1):8-11.
Li CH,
Wang JJ,
Gao LJ,
Wang JH,
Huang ZQ.
Hepatobiliary Surgery Institute, PLA General Hospital, Beijing 100853, China.
OBJECTIVE: To investigate Toll-like receptor 2 (TLR2) and interleukin-1 receptor-associated kinase 1 (IRAK-1) in
bacterial lipoprotein (BLP) tolerance. METHODS: Western blotting was used to confirm the over expression of TLR2
and IRAK-1 in human embryo kidney 293 (HEK293) cells. Plasmids for dual luciferase reporter gene with nuclear
factor-KappaB promoter (pNF-KappaB-Luc) or CMV promoter (phRL-CMV internal control vector) were used to
etect the NF-KappaB activation and the induction of BLP tolerance in HEK-TLR2 cells. RESULTS: BLP stimulation
resulted in dose-dependent NF-KappaB activation in HEK293 cells stably expressing TLR2. And BLP pretreatment
could reduce NF-KappaB activation and induce BLP tolerance in HEK-TLR2 cells. The NF-KappaB activation was
0.329+/-0.010 and 0.168+/-0.010 in BLP-activated and BLP-tolerant HEK-TLR2 cells, respectively. After transfection
with 0.02 mug IRAK-1 plasmid, NF-KappaB activation in the two groups was 0.493+/-0.010 and 0.427+/-0.035, respectively (both P<0.01). So over expression of IRAK-1 could increase NF-KappaB activation in a dose-dependent manner.
CONCLUSION: These results demonstrated that over expression of IRAK-1 could reverse BLP tolerance, whereas over expression of TLR2 failed to prevent the induction of BLP tolerance. Therefore reduced IRAK-1 protein expression is an important mechanism in the development of BLP-induced tolerance, suggesting that it could be a potentially important target for future therapeutic strategies in bacterial infection and sepsis.
http://www.ncbi.nlm.nih.gov/pubmed/20092701
2010,
Toll-like receptor agonists
synergistically increase proliferation
and activation of B cells by
epstein-barr virus.
Epstein-Barr virus
(EBV) efficiently drives proliferation of human primary B cells in vitro, a
process relevant for human diseases such as infectious mononucleosis and
posttransplant lymphoproliferative disease. Human B-cell proliferation is also
driven by ligands of Toll-like receptors (TLRs), notably viral or bacterial DNA
containing unmethylated CpG dinucleotides, which triggers TLR9. Here we
quantitatively investigated how TLR stimuli influence EBV-driven B-cell
proliferation and expression of effector molecules. CpG DNA synergistically
increased EBV-driven proliferation and transformation, T-cell costimulatory
molecules, and early production of interleukin-6. CpG DNA alone activated only
memory B cells, but CpG DNA enhanced EBV-mediated transformation of both memory
and naive B cells. Ligands for TLR2 or TLR7/8 or whole bacteria had a weaker but
still superadditive effect on B-cell transformation. Additionally, CpG DNA
facilitated the release of transforming virus by established EBV-infected
lymphoblastoid cell lines. These results suggest that the proliferation of
EBV-infected B cells and their capability to interact with immune effector cells
may be directly influenced by components of bacteria or other microbes present
at the site of infection.
http://www.ncbi.nlm.nih.gov/pubmed/20089650
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2838115/?tool=pubmed
SEE ALL RELATED:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=20089650
2010, Jan;
Stimulation of axon
regeneration in the mature optic nerve by intravitreal application of the
toll-like receptor 2 agonist Pam3Cys.
PURPOSE:
After injury of the optic nerve,
mature retinal ganglion cells (RGCs) are normally unable to regenerate axons
and undergo apoptosis. However, inflammatory stimulation in the eye induced
by the release of beta/gamma-crystallins from the injured lens or
intravitreal zymosan injection transforms RGCs into an active regenerative
state, protecting these neurons from cell death and allowing them to
regenerate axons back into the optic nerve.
METHODS:
The authors tested whether
intravitreal application of the selective, water-soluble, toll-like receptor
2 agonist Pam(3)Cys can delay axotomized RGC cell death and stimulate the
regeneration of axons using an in vitro and in vivo paradigm.
RESULTS:
Intravitreal injection of
Pam(3)Cys, as lens injury (LI), induced the upregulation of ciliary
neurotrophic factor and glial fibrillary acidic protein expression in
retinal glia accompanied by the activation of the JAK/STAT3 pathway in
RGCs. As a consequence, RGCs switched to a regenerative state, indicated by
a significant upregulation of GAP43 expression and increased neurite
outgrowth of RGCs in culture. Repeated intravitreal Pam(3)Cys application in
vivo induced neuroprotective effects and caused stronger axon regeneration
in the injured optic nerve than observed after LI.
CONCLUSIONS: Pam(3)Cys may be a
suitable agent for stimulating CNS regeneration.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
2010, Feb; Extracellular BCL2
proteins are danger-associated molecular
patterns that reduce tissue damage in
murine models of ischemia-reperfusion
injury.
BACKGROUND: Ischemia-reperfusion (I/R) injury contributes to organ dysfunction in a variety of clinical disorders, including myocardial infarction, stroke, organ transplantation, and hemorrhagic shock. Recent investigations have demonstrated that apoptosis as an important mechanism of cell death leading to organ dysfunction following I/R. Intracellular danger-associated molecular patterns (DAMPs) released during cell death can activate cytoprotective responses by engaging receptors of the innate immune system.
METHODOLOGY/PRINCIPAL FINDINGS: Ischemia was induced in the mouse hind limb by tourniquet or in the heart by coronary artery ligation. Reperfusion injury of skeletal or cardiac muscle was markedly reduced by intraperitoneal or subcutaneous injection of recombinant human (rh)BCL2 protein or rhBCL2-related protein A1 (BCL2A1) (50 ng/g) given prior to ischemia or at the time of reperfusion. The cytoprotective activity of extracellular rhBCL2 or rhBCL2A1 protein was mapped to the BH4 domain, as treatment with a mutant BCL2 protein lacking the BH4 domain was not protective, whereas peptides derived from the BH4 domain of BCL2 or the BH4-like domain of BCL2A1 were. Protection by extracellular rhBCL2 or rhBCL2A1 was associated with a reduction in apoptosis in skeletal and cardiac muscle following I/R, concomitant with increased expression of endogenous mouse BCL2 (mBCL2) protein. Notably, treatment with rhBCL2A1 protein did not protect mice deficient in toll-like receptor-2 (TLR2) or the adaptor protein, myeloid differentiation factor-88 (MyD88).
CONCLUSIONS/SIGNIFICANCE: Treatment with cytokine-like doses of rhBCL2 or rhBCL2A1 protein or BH4-domain peptides reduces apoptosis and tissue injury following I/R by a TLR2-MyD88-dependent mechanism. These findings establish a novel extracellular cytoprotective activity of BCL2 BH4-domain proteins as potent cytoprotective DAMPs.
http://www.ncbi.nlm.nih.gov/pubmed/20161703
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2816997/?tool=pubmed
2010, Mar;
Preconditioning by toll-like receptor 2
agonist Pam3CSK4 reduces CXCL1-dependent leukocyte recruitment in murine
myocardial ischemia/reperfusion injury.
OBJECTIVE:
To test whether preconditioning with a toll-like
receptor (TLR) 2 agonist protects against myocardial ischemia and reperfusion by
interfering with chemokine CXCL1 release from cardiomyocytes.
DESIGN:
C3H mice were challenged with vehicle or
synthetic TLR2 agonist Pam3Cys-Ser-Lys4 (Pam3CSK4; 1 mg/kg) 24 hrs before
myocardial ischemia (20 mins) and reperfusion (2 hrs or 24 hrs). Infarct size,
troponin T release, and leukocyte recruitment were quantified. In murine
cardiomyocytes (HL-1), we studied the expression/activation profile of TLR2 in
response to stimulation with Pam3CSK4 (0.01-1 mg/mL). Furthermore, we studied
the chemokine ligand 1 (CXCL1) response to Pam3CSK4 and ischemia/reperfusion in
vivo and in vitro.
SETTING:
University hospital research laboratory.
SUBJECTS:
Anesthetized male mice and murine
cardiomyocytes.
MEASUREMENTS AND MAIN RESULTS:
Preconditioning by Pam3CSK4 reduced infarct size
and troponin T release. This was accompanied by a decreased recruitment of
leukocytes into the ischemic area and an improved cardiac function. In HL-1
cells, TLR2 activation amplified the expression of the receptor in a
time-dependent manner and led to CXCL1 release in a concentration-dependent
manner. Preconditioning by Pam3CSK4 impaired CXCL1 release in response to a
second inflammatory stimulus in vivo and in vitro.
CONCLUSIONS:
Preconditioning by TLR2 agonist Pam3CSK4 reduces
myocardial infarct size after myocardial ischemia/reperfusion. One of the
mechanisms involved is a diminished chemokine release from cardiomyocytes, which
subsequently limits leukocyte infiltration.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
2010, March;
Immunology and the elusive AIDS vaccine.
Developing a human immunodeficiency virus (HIV) vaccine is
critical to end the global acquired immunodeficiency syndrome (AIDS)
epidemic, but many question whether this goal is achievable. Natural
immunity is not protective, and despite immunogenicity of HIV vaccine
candidates, human trials have exclusively yielded disappointing results.
Nevertheless, there is an indication that success may be possible, but this
will be dependent on understanding the antiviral immune response in
unprecedented depth to identify and engineer the types of immunity required.
Here we outline fundamental immunological questions that need to be answered
to develop a protective HIV vaccine, and the immediate need to harness a
much broader scientific community to achieve this goal.
Reactivation of HIV from latency is regulated by multiple overlapping
factors including chromatinization of the provirus, methylation, T-cell
activation, NFkB
and cytokines including tumour-necrosis factor-a97–99.
Notably, the immune system can regulate the nature of latency in cells
infected with herpesviruses100, and thus the nature of latency is not
necessarily cell intrinsic—apotentially important clue that needs to be
exploited. Latency in macrophages or other cells that may have a role in
persistent infection is less well understood than inTcells.
http://www.ncbi.nlm.nih.gov/pubmed/20220841
2010, March;
Neisseria gonorrhoeae enhances HIV-1
infection of primary resting CD4+ T
cells through TLR2 activation.
Sexually transmitted infections increase the likelihood of HIV-1 transmission. We investigated the effect of Neisseria gonorrheae (gonococcus [GC]) exposure on HIV replication in primary resting CD4(+) T cells, a major HIV target cell during the early stage of sexual transmission of HIV. GC and TLR2 agonists, such as peptidylglycan (PGN), Pam(3)CSK(4), and Pam(3)C-Lip, a GC-derived synthetic lipopeptide, but not TLR4 agonists including LPS or GC lipooligosaccharide enhanced HIV-1 infection of primary resting CD4(+) T cells after viral entry. Pretreatment of CD4(+) cells with PGN also promoted HIV infection. Anti-TLR2 Abs abolished the HIV enhancing effect of GC and Pam(3)C-Lip, indicating that GC-mediated enhancement of HIV infection of resting CD4(+) T cells was through TLR2. IL-2 was required for TLR2-mediated HIV enhancement. PGN and GC induced cell surface expression of T cell activation markers and HIV coreceptors, CCR5 and CXCR4. The maximal postentry HIV enhancing effect was achieved when PGN was added immediately after viral exposure. Kinetic studies and analysis of HIV DNA products indicated that GC exposure and TLR2 activation enhanced HIV infection at the step of nuclear import. We conclude that GC enhanced HIV infection of primary resting CD4(+) T cells through TLR2 activation, which both increased the susceptibility of primary CD4(+) T cells to HIV infection as well as enhanced HIV-infected CD4(+) T cells at the early stage of HIV life cycle after entry. This study provides a molecular mechanism by which nonulcerative sexually transmitted infections mediate enhancement of HIV infection and has implication for HIV prevention and therapeutics.
http://www.ncbi.nlm.nih.gov/pubmed/20147631
2010, May; [Brazilian lyme-like disease
or baggio-yoshinari syndrome: exotic and
emerging brazilian tick-borne zoonosis.]
Lyme disease (LD) is a frequent zoonosis found in the Northern Hemisphere and is considered an infectious disease caused by spirochetes belonging sensu lato to the Borrelia burgdorferi complex transmitted by ticks of the Ixodes ricinus group. In 1992, first cases similar to LD were described in Brazil, when brothers, after a tick bite episode developed symptoms , as erythema migrans, general flu-like symptoms and arthritis. Careful analysis of Brazilian LD-like illness casuistry showed that epidemiological, clinical and laboratorial features in the country were very different from those exhibited by North American and Eurasian LD patients. Human blood-suckers Ixodes ricinus complex ticks were absent at risk areas; the disease is recurrent in the country; Borrelia burgdorferi was never isolated in Brazil and specific serologic tests have shown little positivity with inconsistent results. Furthermore, peripheral blood analysis of patients on electron microscopy exhibited structures resembling Mycoplasma spp, Chlamydia spp and spirochete-like microorganisms. In fact, they were assumed to be latent forms of spirochetes (L form or cell wall deficient bacteria) adapted to survive at inhospitable conditions in vertebrate and invertebrate hosts. For these reasons, the Brazilian zoonosis was named Baggio-Yoshinari Syndrome (BYS) and defined as: "Exotic and emerging Brazilian infectious disease, transmitted by ticks not belonging to the Ixodes ricinus complex, caused by latent spirochetes with atypical morphology, which originates LD-like symptoms, except for occurrence of relapsing episodes and auto-immune disorders".
http://www.ncbi.nlm.nih.gov/pubmed/20676548
Researches conducted at the Laboratory of Investigations in Rheumatology of the Hospital das Clínicas, School of Medicine, Universidade de São Paulo (LIM-17 HCFMUSP) show an occurrence of microorganisms with morphologic structures similar to Mycoplasma spp, Chlamydia spp, and spirochetes without flagella in the peripheral blood of patients with SIRLS, when visualized with Electronic Microscopy (EM).15
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0104-42302010000300025&lng=en&nrm=iso&tlng=en
2010, April;
Primary central nervous system large
B-cell lymphoma with prolific, mixed
T-cell and macrophage infiltrates,
mimicking multiple sclerosis.
Although tissue confirmation is essential for a diagnosis of primary central nervous system large B-cell lymphoma (PCNSBL), accurate assessment may still be difficult, even when tissue is obtained. We report a 59-year-old man, first diagnosed as multiple sclerosis by open biopsy at another institution, who was then correctly diagnosed as PCNSBL after stereotactic biopsy at our hospital. The initial biopsy showed heavy lymphoid and macrophage influx with visible demyelination. On rebiopsy, a diffuse infiltrate of small to medium-sized lymphocytes was prominent and largely stained as T cells (CD3) by immunohistochemistry. There was also an admixture of macrophages, but this time, relatively low numbers of large malignant cells were also identified. The latter stained as B cells (CD20), enabling a diagnosis of B-cell lymphoma, and the condition responded fully to high-dose methotrexate. It is thus possible for PCNSBL to be histologically misinterpreted as a result of ancillary inflammation, characterized here as a profusion of T cells and macrophages.
http://www.ncbi.nlm.nih.gov/pubmed/20425050
AUTISM-RELATED
2010,
May;
Exposure of Toll-like receptors 4 to
bacterial lipopolysaccharide (LPS)
impairs human colonic smooth muscle cell
function.
Endotoxemia by bacterial lipopolysaccharide (LPS) has been reported to affect gut motility specifically depending on Toll-like receptor 4 activation (TLR4). However, the direct impact of LPS ligation to TLR4 on human smooth muscle cells (HSMC) activity still remains to be elucidated. The present study shows that TLR4, its associated molecule MD2, and TLR2 are constitutively expressed on cultured HSMC and that, once activated, they impair HSMC function. The stimulation of TLR4 by LPS induced a time- and dose-dependent contractile dysfunction, which was associated with a decrease of TLR2 messenger, a rearrangement of microfilament cytoskeleton and an oxidative imbalance, i.e., the formation of reactive oxygen species (ROS) together with the depletion of GSH content. An alteration of mitochondria, namely a hyperpolarization of their membrane potential, was also detected. Most of these effects were partially prevented by the NADPH oxidase inhibitor apocynin or the NFkappaB inhibitor MG132. Finally, a 24 h washout in LPS-free medium almost completely restored morphofunctional and biochemical HSMC resting parameters, even if GSH levels remained significantly lower and no recovery was observed in TLR2 expression. Thus, the exposure to bacterial endotoxin directly and persistently impaired gastrointestinal smooth muscle activity indicating that HSMC actively participate to dysmotility during infective burst. The knowledge of these interactions might provide novel information on the pathogenesis of infection-associated gut dysmotility and further clues for the development of new therapeutic strategies.
http://www.ncbi.nlm.nih.gov/pubmed/20112289
Yale's Congenital Lyme/Syphilis Brain
Damage Explained by the NIH (It's
Pam3Cys or LYMErix):
2010, Jul, NIH;
TLR2 activation
inhibits embryonic neural progenitor
cell proliferation.
Toll-like receptors (TLRs) play essential roles in innate immunity, and increasing evidence indicates that these receptors are expressed in neurons, astrocytes, and microglia in the brain, where they mediate responses to infection, stress, and injury. To address the possibility that TLR2 heterodimer activation could affect progenitor cells in the developing brain, we analyzed the expression of TLR2 throughout mouse cortical development, and assessed the role of TLR2 heterodimer activation in neuronal progenitor cell (NPC) proliferation. TLR2 mRNA and protein was expressed in the cortex in embryonic and early postnatal stages of development, and in cultured cortical NPC. While NPC from TLR2-deficient and wild type embryos had the same proliferative capacity, TLR2 activation by the synthetic bacterial lipopeptides Pam(3)CSK(4) and FSL1, or low molecular weight hyaluronan, an endogenous ligand for TLR2, inhibited neurosphere formation in vitro. Intracerebral in utero administration of TLR2 ligands resulted in ventricular dysgenesis characterized by increased ventricle size, reduced proliferative area around the ventricles, increased cell density, an increase in phospho-histone 3 cells, and a decrease in BrdU(+) cells in the sub-ventricular zone. Our findings indicate that loss of TLR2 does not result in defects in cerebral development. However, TLR2 is expressed and functional in the developing telencephalon from early embryonic stages and infectious agent-related activation of TLR2 inhibits NPC proliferation. TLR2-mediated inhibition of NPC proliferation may therefore be a mechanism by which infection, ischemia, and inflammation adversely affect brain development.
http://www.ncbi.nlm.nih.gov/pubmed/20456021
2010; Chinese grow stealth,
antibiotic-resistant mycoplasma:
2010, July, Mutations in 23S rRNA
gene associated with decreased
susceptibility to tiamulin and
valnemulin in Mycoplasma gallisepticum.
Mycoplasma gallisepticum is a major etiological agent of chronic respiratory disease (CRD) in chickens and sinusitis in turkeys. The pleuromutilin antibiotics tiamulin and valnemulin are currently used in the treatment of M. gallisepticum infection. We studied the in vitro development of pleuromutilin resistance in M. gallisepticum and investigated the molecular mechanisms involved in this process. Pleuromutilin-resistant mutants were selected by serial passages of M. gallisepticum strains PG31 and S6 in broth medium containing subinhibitory concentrations of tiamulin or valnemulin. A portion of the gene encoding 23S rRNA gene (domain V) and the gene encoding ribosome protein L3 were amplified and sequenced. No mutation could be detected in ribosome protein L3. Mutations were found at nucleotide positions 2058, 2059, 2061, 2447 and 2503 of 23S rRNA gene (Escherichia coli numbering). Although a single mutation could cause elevation of tiamulin and valnemulin MICs, combinations of two or three mutations were necessary to produce high-level resistance. All the mutants were cross-resistant to lincomycin, chloramphenicol and florfenicol. Mutants with the A2058G or the A2059G mutation exhibited cross-resistance to macrolide antibiotics erythromycin, tilmicosin and tylosin.
http://www.ncbi.nlm.nih.gov/pubmed/20487023
2010, Aug, Clifford
Harding;
Mycobacterium tuberculosis and TLR2
agonists inhibit induction of type I IFN
and class I MHC antigen cross processing
by TLR9.
Dendritic cells (DCs) cross process exogenous Ags and present them by class I MHC (MHC-I) molecules to CD8(+) T cells specific for Ags from viruses and bacteria such as Mycobacterium tuberculosis. Unmethylated CpG DNA signals through TLR9 to induce type I IFN (IFN-alpha/beta), which enhances MHC-I Ag cross processing, but lipoproteins that signal through TLR2 do not induce IFN-alpha/beta. In these studies we observed that M. tuberculosis, which expresses agonists of both TLR9 and TLR2, did not induce production of IFN-alpha/beta or cross processing by murine DCs. Furthermore, M. tuberculosis and TLR2 agonists inhibited induction of IFN-alpha/beta and DC cross processing by CpG DNA. Exogenous IFN-alpha/beta effectively enhanced cross processing of M. bovis bacillus Calmette-Guérin expressing OVA, bypassing the inhibition of induction of endogenous IFN-alpha/beta. In addition, inhibition of TLR9-induced cross processing of M. bovis bacillus Calmette-Guérin expressing OVA could be circumvented by pretreating cells with CpG DNA to induce IFN-alpha/beta and MHC-I cross processing before inhibitory mycobacterial TLR2 agonists were present. Inhibition of the response to one TLR by another may affect the ultimate response to pathogens like M. tuberculosis that express agonists of multiple TLRs, including TLR2 and TLR9. This mechanism may contribute to immune evasion and explain why IFN-alpha/beta provides little contribution to host immunity to M. tuberculosis. However, downregulation of certain TLR responses may benefit the host by preventing detrimental excessive inflammation that may occur in the presence of persistent infection.
http://www.ncbi.nlm.nih.gov/pubmed/20660347
2010, Aug, Wormser, Klempner, Latov;
Anti-neural antibody reactivity in
patients with a history of Lyme
borreliosis and persistent symptoms.
Some Lyme disease patients report debilitating chronic symptoms of pain, fatigue, and cognitive deficits despite recommended courses of antibiotic treatment. The mechanisms responsible for these symptoms, collectively referred to as post-Lyme disease syndrome (PLS) or chronic Lyme disease, remain unclear. We investigated the presence of immune system abnormalities in PLS by assessing the levels of antibodies to neural proteins in patients and controls. Serum samples from PLS patients, post-Lyme disease healthy individuals, patients with systemic lupus erythematosus, and normal healthy individuals were analyzed for anti-neural antibodies by immunoblotting and immunohistochemistry. Anti-neural antibody reactivity was found to be significantly higher in the PLS group than in the post-Lyme healthy (p<0.01) and normal healthy (p<0.01) groups. The observed heightened antibody reactivity in PLS patients could not be attributed solely to the presence of cross-reactive anti-borrelia antibodies, as the borrelial seronegative patients also exhibited elevated anti-neural antibody levels. Immunohistochemical analysis of PLS serum antibody activity demonstrated binding to cells in the central and peripheral nervous systems. The results provide evidence for the existence of a differential immune system response in PLS, offering new clues about the etiopathogenesis of the disease that may prove useful in devising more effective treatment strategies. http://www.ncbi.nlm.nih.gov/pubmed/20227484
2010, Aug;
Impaired Functions of Peripheral Blood
Monocyte Subpopulations in Aged Humans.
Aging is associated with increased susceptibility to microbial infections, and
monocytes play an important role in microbial defense. In this study, we have
identified and compared four subpopulations of monocytes (CD14(++(high))CD16(-),
CD14(+(low))CD16(-), CD14(++(high))CD16(+), and CD14(+(low))CD16(+)) in the
peripheral blood of young and aged subjects with regard to their numbers,
cytokine production, TLR expression, and phosphorylation of ERK1/2 in response
to pam3Cys a TLR-1/2 ligand. Proportions and numbers of
CD14(++(high))CD16(+) and CD14(+(low))CD16(+) monocytes were significantly
increased, whereas proportions of CD14(+(low))CD16(-) monocytes were decreased
in aged subjects as compared to young subjects. In aged subjects, IL-6
production by all four subsets of monocytes was significantly decreased, whereas
TNF-alpha production was decreased in monocyte subsets, except the
CD14(+(low))CD16(-) subset. A significantly reduced expression of TLR1 was
observed in CD14(++(high))CD16(+) and CD14(+(low))CD16(+) monocyte subsets in
aged subjects. Furthermore, following pam3Cys stimulation, ERK1/2
phosphorylation was significantly lower in CD14(+(low))CD16(+),
CD14(++(high))CD16(+), and CD14(+(low))CD16(-) subsets of monocytes from aged
subjects. This is the first study of four subpopulations of monocytes in aging,
which demonstrates that their functions are differentially impaired with regard
to the production of cytokines, expression of TLR, and signaling via the
ERK-MAPK pathway. Finally, changes in the number of monocyte subsets, and
impairment of TLR1 expression, TNF-alpha production, and EK1/2 phosphorylation
was more consistent in CD16(+) monocyte subsets regardless of expression of
CD14(high) or CD14(+low), therefore highlighting the significance of further
subdivision of monocytes into four subpopulations.
http://www.ncbi.nlm.nih.gov/pubmed/19661221
2010, Aug;
The synthetic bacterial lipopeptide Pam3CSK4 modulates
respiratory syncytial virus infection independent of TLR activation.
Respiratory syncytial virus (RSV) is an
important cause of acute respiratory disease in infants, immunocompromised
subjects and the elderly. However, it is unclear why most primary RSV
infections are associated with relatively mild symptoms, whereas some result
in severe lower respiratory tract infections and bronchiolitis. Since RSV
hospitalization has been associated with respiratory bacterial
co-infections, we have tested if bacterial Toll-like receptor (TLR) agonists
influence RSV-A2-GFP infection in human primary cells or cell lines. The
synthetic bacterial lipopeptide Pam3-Cys-Ser-Lys4 (Pam3CSK4), the prototype
ligand for the heterodimeric TLR1/TLR2 complex, enhanced RSV infection in
primary epithelial, myeloid and lymphoid cells. Surprisingly,
enhancement was optimal when lipopeptides and virus were added
simultaneously, whereas addition of Pam3CSK4 immediately after infection had
no effect. We have identified two structurally related lipopeptides without
TLR-signaling capacity that also modulate RSV infection, whereas
Pam3CSK4-reminiscent TLR1/2 agonists did not, and conclude that modulation
of infection is independent of TLR activation. A similar TLR-independent
enhancement of infection could also be demonstrated for wild-type RSV
strains, and for HIV-1, measles virus and human metapneumovirus. We show
that the effect of Pam3CSK4 is primarily mediated by enhanced binding of RSV
to its target cells. The N-palmitoylated cysteine and the cationic lysines
were identified as pivotal for enhanced virus binding. Surprisingly, we
observed inhibition of RSV infection in immortalized epithelial cell lines,
which was shown to be related to interactions between Pam3CSK4 and
negatively charged glycosaminoglycans on these cells, which are known
targets for binding of laboratory-adapted but not wild-type RSV. These data
suggest a potential role for bacterial lipopeptides in enhanced binding of
RSV and other viruses to their target cells, thus affecting viral entry or
spread independent of TLR signaling. Moreover, our results also suggest a
potential application for these synthetic lipopeptides as adjuvants for
live-attenuated viral vaccines. http://www.ncbi.nlm.nih.gov/pubmed/20808895
2010, Sep,
Coinfection with EBV/CMV and other respiratory agents in
children with suspected infectious mononucleosis.
BACKGROUND: Numerous studies
have shown that Epstein-Barr virus (EBV) and cytomegalovirus (CMV) can
infect immunocompetent patients simultaneously with other agents.
Nonetheless, multiple infections with other agents in EBV/CMV-infected
children have received little attention. We conducted a retrospective study
of children with suspected infectious mononucleosis. Peripheral blood
samples were analyzed by indirect immunofluorescence to detect EBV, CMV and
other respiratory agents including respiratory syncytial virus; adenovirus;
influenza virus types A and B; parainfluenza virus types 1, 2 and 3;
Chlamydia pneumoniae and Mycoplasma pneumoniae. A medical history was
collected for each child.
RESULTS: The occurrence of
multipathogen infections was 68.9%, 81.3% and 63.6% in the children with
primary EBV, CMV or EBV/CMV, respectively, which was significantly higher
than that in the past-infected group or the uninfected group (p < 0.001). Of
the multipathogen-infected patients, the incidence of C. pneumoniae in
children with primary infection was as high as 50%, significantly higher
than in the other groups (p < 0.001). In the patients with multipathogen
infection and EBV/CMV primary infection, fever, rash, lymphadenopathy,
hepatomegaly, splenomegaly, atypical lymphocytes and abnormal liver function
were more frequent and the length of hospital stay and duration of fever
were longer than in other patients.
CONCLUSION: Our study suggests
that there is a high incidence of multipathogen infections in children
admitted with EBV/CMV primary infection and that the distribution of these
pathogens is not random.
http://www.ncbi.nlm.nih.gov/pubmed/20858235
2010, Sep, Clifford
Harding;
Mycobacterium tuberculosis promotes HIV
trans-infection and suppresses major histocompatibility complex class II
antigen processing by dendritic cells.
Mycobacterium tuberculosis is a leading killer of HIV-infected individuals worldwide, particularly in sub-Saharan Africa, where it is responsible for up to 50% of HIV-related deaths. Infection by HIV predisposes individuals to M. tuberculosis infection, and coinfection accelerates the progression of both diseases. In contrast to most other opportunistic infections associated with HIV, an increased risk of M. tuberculosis infection occurs during early-stage HIV disease, long before CD4 T cell counts fall below critical levels. We hypothesized that M. tuberculosis infection contributes to HIV pathogenesis by interfering with dendritic cell (DC)-mediated immune control. DCs carry pathogens like M. tuberculosis and HIV from sites of infection into lymphoid tissues, where they process and present antigenic peptides to CD4 T cells. Paradoxically, DCs can also deliver infectious HIV to T cells without first becoming infected, a process known as trans-infection. Lipopolysaccharide (LPS)-activated DCs sequester HIV in pocketlike membrane invaginations that remain open to the cell surface, and individual virions are delivered from the pocket into T cells at the site of contact during trans-infection. Here we report that M. tuberculosis exposure increases HIV trans-infection and induces viral sequestration within surface-accessible compartments identical to those seen in LPS-stimulated DCs. At the same time, M. tuberculosis dramatically decreases the degradative processing and major histocompatibility complex class II (MHC-II) presentation of HIV antigens to CD4 T cells. Our data suggest that M. tuberculosis infection promotes a shift in the dynamic balance between antigen processing and intact virion presentation, favoring DC-mediated amplification of HIV infections.
http://www.ncbi.nlm.nih.gov/pubmed/20592078
2010,
Sep; Roles of
Toll-like receptor 2 (TLR2) and
superantigens on adaptive immune
responses during CNS staphylococcal
infection.
Staphylococcus aureus is a common etiologic agent of brain abscesses and possesses numerous virulence factors that manipulate host immunity. One example is superantigens (SAG) that clonally expand T cell subsets bearing specific Vβ receptors. Toll-like receptor 2 (TLR2) is one receptor implicated in S. aureus recognition. However, the interplay between TLR2, SAG, and adaptive immunity during brain abscess formation has not yet been investigated and could reveal novel insights into host-pathogen interactions for regulating protective immunity. A comprehensive analysis of abscess-associated T cell populations in TLR2 KO and WT mice was performed following infection with a S. aureus clinical isolate. Both natural killer T (NKT) and γδ T cell infiltrates were increased in brain abscesses of TLR2 KO mice and produced more IL-17 and IFN-γ compared to WT populations, which could have resulted from elevated bacterial burdens observed in these animals. Analysis of SAG-reactive T cells revealed a predominant Vβ(8.1,8.2) infiltrate reactive with staphylococcal enterotoxin B (SEB), whereas SEA-reactive Vβ(11) T cells were less numerous. Brain abscesses of TLR2 KO mice had fewer Vβ(8.1,8.2) and Vβ(11) T cells and produced less TNF-α and IFN-γ compared to WT animals. Treatment of primary microglia with purified SEB augmented TNF-α production in response to the TLR2 ligand Pam3Cys, which may serve to amplify proinflammatory cascades during CNS S. aureus infection. Collectively, these studies demonstrate that TLR2 impacts adaptive immunity to S. aureus infection and modulates SAG responses.
http://www.ncbi.nlm.nih.gov/pubmed/20868736
2010, Oct;
Accurate and efficient gp120 V3 loop structure based models for
the determination of HIV-1 co-receptor usage
Background
HIV-1 targets human cells expressing both the CD4 receptor, which binds the
viral envelope glycoprotein gp120, as well as either the CCR5 (R5) or CXCR4
(X4) co-receptors, which interact primarily with the third hypervariable
loop (V3 loop) of gp120. Determination of HIV-1 affinity for either the R5
or X4 co-receptor on host cells facilitates the inclusion of co-receptor
antagonists as a part of patient treatment strategies. A dataset of 1193
distinct gp120 V3 loop peptide sequences (989 R5-utilizing, 204 X4-capable)
is utilized to train predictive classifiers based on implementations of
random forest, support vector machine, boosted decision tree, and neural
network machine learning algorithms. An in silico mutagenesis procedure
employing multibody statistical potentials, computational geometry, and
threading of variant V3 sequences onto an experimental structure, is used to
generate a feature vector representation for each variant whose components
measure environmental perturbations at corresponding structural positions.
Results
Classifier performance is evaluated based on stratified 10-fold
cross-validation, stratified dataset splits (2/3 training, 1/3 validation),
and leave-one-out cross-validation. Best reported values of sensitivity
(85%), specificity (100%), and precision (98%) for predicting X4-capable
HIV-1 virus, overall accuracy (97%), Matthew's correlation coefficient
(89%), balanced error rate (0.08), and ROC area (0.97) all reach critical
thresholds, suggesting that the models outperform six other state-of-the-art
methods and come closer to competing with phenotype assays.
Conclusions
The trained classifiers provide instantaneous and reliable predictions
regarding HIV-1 co-receptor usage, requiring only translated V3 loop
genotypes as input. Furthermore, the novelty of these computational
mutagenesis based predictor attributes distinguishes the models as
orthogonal and complementary to previous methods that utilize sequence,
structure, and/or evolutionary information. The classifiers are available
online at http://proteins.gmu.edu/automute
http://www.biomedcentral.com/content/pdf/1471-2105-11-494.pdf
???
We don't know what ▲that is. We don't know what HIV gp120 is, and
neither does the head of the NIAID, Anthony Fauci. All we know is that
Pam3Cys as an HIV vaccine analog/adjuvant could not and did not work.
2010, Dec, Iran: Seroprevalence of human herpes simplex, hepatitis B and epstein-barr viruses in children with acute lymphoblastic leukemia in southern Iran.
Mahjour SB, Ghaffarpasand F, Fattahi MJ, Ghaderi A, Fotouhi Ghiam A, Karimi M.
Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract
To investigate the seroprevalence of Herpes Simplex Viruses (HSV1 and HSV2), Ebstein-Barr Virus (EBV) and Hepatitis B Virus (HBV) in children with acute lymphoblastic leukemia (ALL) in southern Iran. 90 patients with ALL and 90 age-sex matched healthy participants were enrolled in this study. Antibodies (IgG) against HBsAg, HSV1, HSV2, EBV different antigens including Epstein-Barr nuclear antigen-1 (EBNA-1), viral capsid antigen (VCA) and early antigen (EA) were measured by enzyme-linked immunosorbent assay (ELISA). There were 54 (60%) male and 36 (40%) female in both study groups with mean age of 8.47 ± 3.61 and 8.61 ± 2.84 years in case and control group respectively (P = 0.812). The prevalence of antibodies against HBsAg (P = 0.002), HSV1 (P < 0.0001), VCA (P = 0.021) and EA (P < 0.0001) antigens of EBV were significantly higher in ALL patients. With the results of this study, we could not exclude a connection between these viral infections and later leukemogenesis in childhood ALL, although nor latent infection nor congenital infection cannot be excluded by this method.
http://www.ncbi.nlm.nih.gov/pubmed/20309661
2010, Oct;
Interactions between bacterial pathogens
and mitochondrial cell death pathways.
"The modulation of host cell death pathways by bacteria has been recognized as a major pathogenicity mechanism. Among other strategies, bacterial pathogens can hijack the cell death machinery of host cells by influencing the signalling pathways that converge on the mitochondria. In particular, many bacterial proteins have evolved to interact in a highly specific manner with host mitochondria, thereby modulating the decision between cell life and death. In this Review, we explore the intimate interactions between bacterial pathogens and mitochondrial cell death pathways."
http://www.ncbi.nlm.nih.gov/pubmed/20818415
http://www.nature.com/nrmicro/journal/v8/n10/full/nrmicro2421.html
2010, Oct, Brazil; Mycoplasmal
lipid-associated membrane proteins and
Mycoplasma arthritidis mitogen
recognition by serum antibodies from
patients with rheumatoid arthritis.
Mycoplasmal lipid-associated membrane proteins (LAMPs) and Mycoplasma arthritidis mitogen (MAM superantigen) are potent stimulators of the immune system. The objective of this work was to detect antibodies to MAM and LAMPs of Mycoplasma hominis and M. fermentans in the sera of patients affected by rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) to identify mycoplasmal products that can be involved in the etiopathogenesis of these autoimmune diseases. Serum samples from female RA and SLE patients and controls, recombinant MAM, and LAMPs of M. hominis PG21 and M. fermentans PG18 were used in Western blot assays. A similar frequency of sera from patients and controls reactive to MAM was detected. A larger number of M. hominis and M. fermentans LAMPs were recognized by sera from RA patients than controls, but no differences were detected between sera from SLE patients and controls. Among the LAMPs recognized by IgG antibodies from RA patients, proteins of molecular masses in a range of <49 and ≥20 KDa (M. hominis) and <102 and ≥58 KDa (M. fermentans) were the most reactive. These preliminary results demonstrate the strong reactivity of antibodies of RA patients with some M. hominis and M. fermentans LAMPs. These LAMPs could be investigated as mycoplasmal antigens that can take part in the induction or amplification of human autoimmune responses.
http://www.ncbi.nlm.nih.gov/pubmed/21052674
2010, Nov;
MULTICOMPONENT MORAXELLA CATARRHALIS
OUTER MEMBRANE VESICLES INDUCE AN
INFLAMMATORY RESPONSE AND ARE
INTERNALIZED BY HUMAN EPITHELIAL CELLS.
Moraxella catarrhalis is an emerging human respiratory pathogen in patients with chronic obstructive pulmonary disease (COPD) and in children with acute otitis media. The specific secretion machinery known as outer membrane vesicles (OMV) is a mechanism by which Gram negative pathogens interact with host cells during infection. We identified 57 proteins in M. catarrhalis OMV using a proteomics approach combining 2-dimensional SDS-PAGE and MALDI-TOF mass spectrometry analysis. The OMV contained known surface proteins such as ubiquitous surface proteins (Usp) A1/A2, and Moraxella IgD-binding protein (MID). Most of the proteins are adhesins/ virulence factors triggering the immune response, but also aid bacteria to evade the host defense. FITC-stained OMV bound to lipid raft domains in alveolar epithelial cells and were internalized after interaction with Toll-like receptor 2 (TLR2) suggesting a delivery to the host tissue of a large and complex group of OMV-attributed proteins. Interestingly, OMV modulated the pro-inflammatory response in epithelial cells, and UspA1-bearing OMV were found to specifically down-regulate the reaction. When mice were exposed to OMV, a pulmonary inflammation was clearly seen. Our findings indicate that Moraxella OMV are highly biologically active, transport main bacterial virulence factors, and may modulate the epithelial pro-inflammatory response.
http://www.ncbi.nlm.nih.gov/pubmed/21044239
2010, Dec, Clifford
Harding; Mycobacterium tuberculosis synergizes
with ATP to induce release of microvesicles and exosomes containing
major histocompatibility complex class
II molecules capable of antigen
presentation.
Major histocompatibility complex class II (MHC-II) molecules are released by murine macrophages upon lipopolysaccharide (LPS) stimulation and ATP signaling through the P2X7 receptor. These studies show that infection of macrophages with Mycobacterium tuberculosis or M. bovis strain BCG enhances MHC-II release in synergy with ATP. Shed MHC-II was contained in two distinct organelles, exosomes and plasma membrane-derived microvesicles, which were both able to present exogenous antigenic peptide to T hybridoma cells. Furthermore, microvesicles from mycobacterium-infected macrophages were able to directly present M. tuberculosis antigen (Ag) 85B(241-256)-I-A(b) complexes that were generated by the processing of M. tuberculosis Ag 85B in infected cells to both M. tuberculosis-specific T hybridoma cells and naïve P25 M. tuberculosis T-cell receptor (TCR)-transgenic T cells. In the presence of prefixed macrophages, exosomes from mycobacterium-infected macrophages provided weak stimulation to M. tuberculosis-specific T hybridoma cells but not naïve P25 T cells. Thus, infection with M. tuberculosis primes macrophages for the increased release of exosomes and microvesicles bearing M. tuberculosis peptide-MHC-II complexes that may generate antimicrobial T-cell responses.
http://www.ncbi.nlm.nih.gov/pubmed/20837713
Steere's ▲ kinda Lyme?? The complex, itself, because of binding kinetics, creates a new antigen and is shed for that reason? The complex - especially when it's Allen Steere's HLA set - is toxic?
20101226; NOTES TO SELF: Previous notes are at the bottom of this page, for whoever is interested.
News: Johns Hopkins Autism and the Pediatric Journal that reveals the problems with vaccines (expansions of virus and mutations of them).
1) Johns Hopkins MD whose daughter won an Autism lawsuit against vaccine mfgs. = Fungal-Viral Synergy:
"bacterial pathogens can hijack the cell death machinery of host cells by influencing the signalling pathways that converge on the mitochondria." http://www.nature.com/nrmicro/journal/v8/n10/full/nrmicro2421.html
2) "Vaccination Causes Mutations" - Pediatric Journal
"This analysis demonstrates that additional antiviral resistance can rapidly develop in a previously single-resistant strain as a result of mutation, ***drug response or gene exchange with another virus,*** the researchers wrote. "
"According to the researchers, immunocompromised [Johns Hopkins Autism and LYMErix Disease clue] patients were more susceptible to the emergence of oseltamivir-resistant H1N1 virus on treatment and also transmitted the virus to others, despite often having no influenza symptoms or having completed antiviral therapy."
3) A treatment of severe rheumatoid arthritis is TNF-inhibitors. People with RA taking TNF-inhibitors are susceptible to opportunistic infections - just like LYMErix Disease victims. One of the mechanisms of Pam3Cys or LYMErix Disease (un-inhibition of latent Opportunistics and tolerance to unusual fungal lipids, Tlr2/1 agonists, mycoplasma/mycobacteria) is also the inhibition of TNF production. (Search for TNF in this page.)
http://www.ncbi.nlm.nih.gov/pubmed?term=pam3cys[All%20Fields]%20AND%20tnf[All%20Fields]&cmd=DetailsSearch
4) Brazil (scroll down to May, 2010) scientists and Garth Nicolson make these same observations.
5) Birds carry Borrelia, Influenza and their own version of a Tuberculosis. You get what that means.
http://www.ncbi.nlm.nih.gov/pubmed/18798030
6) Bioweaponeer Joe Tully also revealed to us that there are spirosplasmas etc in ticks.
7) Complications of Sepsis - Streptococcal infections and TLR2:
http://www.ncbi.nlm.nih.gov/pubmed/20946179
8) The Same ▲ Strep Dysimmunity is also related to ADHD. (You know, Chronic Strep, the Fidget-Disease.)
1) New Chinese testing proposal: Use band 41 and each other band is as accurate for diagnosis as its assigned percent specificity: China-Lyme Reverse Breakthru on Diagnosis
2) "Immature B-cells and Mycoplasma" (one result from the search):
"In contrast to the polyclonal B cell mitogen LPS, they were unable to promote Ig isotype switching." (OspA renders immune-incompetent):
http://www.ncbi.nlm.nih.gov/pubmed?term=17698916[uid]&cmd=DetailsSearch
3) IMMATURE B-CELLS AND EPSTEIN-BARR:
http://www.ncbi.nlm.nih.gov/pubmed?term=%28%22precursor%20cells%2C%20b-lymphoid%22[
LINK REMOVED]
4) http://www.ncbi.nlm.nih.gov/pubmed/9341792?dopt=Abstract (fungal anti-apoptosis)
See All Related and sort by ▲date.
5) JPMorgan Chase just sent Chinese Medicine $69 million dollars in investment cash because they
happen to tell the *TRUTH* whereas the CT AG had to *sue* them [IDSociety.org] and they *STILL*
haven't cooperated,... [BLUMENTHAL_IDSA_FEB_2010.pdf] ... while UConn begs for $$ to refurbish
that hell hole of a hospital... (100411)
Almost nothing happened in the 1990s (See DeFoort, Koreans and HIV gp120/Pam3Cys).
Why?
The confluence of Yale lying about OspA vaccines and the NIH's shift to focus on HIV because of the high frequency of mycoplasma and mycobacteria (Tuberculosis) and Epstein-Barr that occurs in AIDS:
All along a primary culprit in all these outcomes was Pam3Cys or the synthetic Braun lipoprotein or OspA, which, amazingly ended up on both vaccines, despite not ever working as Tuberculosis vaccines alone.
Yale had decided to lie about vaccines outcomes ahead of time, which is why Allen Steere went to Europe in 1992 to falsify the diagnostic standard (leaving OspA and B out deliberately, for the post-LYMErix-FDA-approval era and the monopoly on blood, as explained by Robert Schoen in 1998 and Schoen and Persing in their 1995 RICO patent).
We think it is CERTAIN that Yale looked at the blood of recombinant OspA-vaccinated mice and primates in the early 1990s and found the Western Blots to be unreadable because in no studies - not human and not animal studies - have they ever produced a Western Blot from early trials (Phase I and II) and they always assessed vaccines outcomes on the basis of arthritis in the joints of animals. And they always CLAIMED that these vaccines did not produce protective antibodies for humans, but rather, worked by disinfecting ticks. If you can believe it.
One of the "studies" where bogus science was applied, was the "immunoflourescing antibodies" experiment where the Lyme Mafia - despite having a valid DNA method at the ready in 1991 and for which they applied for a patent in 1993 - Bb specific flagellin (See Chapter 1 of CRYME DISEASE) - disclaims itself as an advertisement:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC49303/pdf/pnas01086-0226.pdf
There is not ▲going to be any of the vaccine-brand OspA in tick-mash because of selection pressure and there will be no fluorescing anti-flagellin because flagellin are internal.
These crooks knew their OspA vaccines weren't going to work so they just made up a bunch of crazy shit and shoved it down everyone's throat with a healthy dose of You-Have-Lacka-Wacka-Pee-Pee-itis, which gave psychiatrists something to do because real science has been squeezing their Blame-The-Victim playground for the last 5 decades.
6) http://www.ncbi.nlm.nih.gov/pubmed?term=%22chronic%20epstein-barr%22[All%20Fields]&cmd=DetailsSearch
7) http://www.ncbi.nlm.nih.gov/pubmed?term=%28%22fatigue%20syndrome%2C%20chronic%22[MeS///{LINK REMOVED, AND MYCOPLASMA]
8)
http://www.ncbi.nlm.nih.gov/pubmed?term=nicolson[All%20Fields]%20AND%20%28%22mycoplasma%22[ LINK REMOVED, NICOLSON]
“To be most effective, advances in regulatory science must be fully
integrated into the entire product development process.”
Says the new FDA chief, Margaret
Hamburg:
http://www.fda.gov/ScienceResearch/SpecialTopics/RegulatoryScience/ucm228206.htm
Which is the End of the Almighty Checklist - a sign of the
auto-detonation of the adherents to Psychiatry:
The Biomarkers published
by the same gang who now says Lyme is a consequence of inadequate sexual
release. After all, the head of the FDA now recommends that we do:
1)
Using Dark Field to study the
modified erythrocytes, modified via the tolerance induced from chronic
stimulation of TLR2 by spirochetal blebbing of the likes of TLR2 agonist OspA,
and subsequent colonization by mycoplasma (recall that many sufferers of
“Chronic Fatigue Syndrome” have been found to own a colony of Candida (a
fungus).
2)
DNA/RNA of all the mycoplasma and
activated viral infections that won't be producing antibodies (because some are also TLR2 agonists
like Epstein-Barr, and chronic TLR2 agonism results in no-antibodies (Justin
Radolf: ,” Inhibition of MHC-II
Ag processing by either MTB bacilli or purified MTB 19-kDa
lipoprotein was dependent on Toll-like receptor (TLR) 2 and
independent of TLR 4. Synthetic analogs of lipopeptides from
Treponema pallidum also inhibited Ag processing. Despite the ability
of MTB 19-kDa lipoprotein to activate microbicidal and innate immune
functions early in infection, TLR 2-dependent inhibition of MHC-II
expression and Ag processing by MTB 19-kDa lipoprotein during later
phases of macrophage infection may prevent presentation of MTB Ags
and decrease recognition by T cells.”
http://www.jimmunol.org/cgi/content/full/167/2/910 )]
3)
QEEG
or Lenny Sigal’s findings that persons with chronic Lyme have changes to
electroencephalograms.
http://www.ncbi.nlm.nih.gov/pubmed/7554300 “Abnormal QEEG and/or EPs were
found in 75% of the active Lyme disease patients and in 54% of the post CNS Lyme
disease patients.”
4)
modified cytokine profile
suggested by Lenny Sigal, in the Cold Spring Harbor book mentioned above,
to which Paul Duray attended and re-iterated his “EBV-modified lymphocytes”
findings:
page 205: Says Sigal: “Since cytokines can alter endothelial cell function and
increase the entry of inflammatory cells to the organ whose vasculature has been
modified, the presence of cytokines in the Lyme synovium may have an important
indirect effect on local inflammation [Yednock et al. 1992]. The presences of
circulating levels of these cytokines may be the cause of the certain clinical
features of Lyme disease; eg., sleep disorder due to elevated levels of IL-1
(Opp and Krueger 1991) may be part f the cause of fibromyalgia seen during and
after active Lyme disease (Sigal 1990).”
5)
MMP-130 and GFAp in the CSF
of Mark Klempner's
Neuroborreliosis (which is not "Lyme Disease," BTW,
because Lyme Disease became "only the HLA-linked knee-presentation" at
Dearborn),
6)
Allen Steere's
Nitric Oxide in the CSF of Lyme victims (we now know that the exposure to
these reactive oxygen species/byproducts are downregulators of TLR2)
http://www.ncbi.nlm.nih.gov/pubmed/7513330
7)
CORRECT Borrelia (all species) DNA/RNA
primers and not the OspA gene,
because that changes according to
Alan Barbour; we can throw out all "studies" in which the
OspA gene was the determinant for infection with the "Lyme" spirochete
8)
Quinolinic acid
and the degradants of monoamines in the CSF of borreliosis victims (JJ
Halperin),
9)
brain SPECT and PET imaging
(we now know that the metabolism of erythrocytes are hijacked by the mycoplasma
in the blood - infections we can't fight off because they have OspA or TLR2
agonizing lipoproteins in them), NOTE: psychiatry
likes to suggest that changes to the brain perfusion seen in chronic
Lyme/Fatigue victims is a result of Bipolar, when not ever once was anyone who
was diagnosed with Bipolar have shown hyper- or hypo- perfusion as a result of
any of the “poles” without other true, real, scientific valid biomarkers of real
illness first performed as a rule out. Secondarily, when studies have been
performed in which there are brain structural changes to the likes of 17 year
olds who are determined to be psychopathic or sociopathic, have these conditions
studied a contribution by trauma. As a third outstanding parameter, why, we
wonder, are lawyers undable to view anything from a scientific standpoint? Why
do they score so low on the visual-spatial scale of cognitive abilities? Why
are lawyers so left-brain dominant and why do they have no understanding of
vocabulary as a simple means to draw a picture in the mind of another.
Most of all, why is this seen in both lawyers and psychiatrists? Why are the
best scientists and engineers known to be visual-spatial or right-brain
dominant? Is this nature or nurture? The victims of their chronic fraud crime
are not entitled to know why self-alleged scholars as these are clearly unable
to think. We are not entitled to see their own cognitive differentials and
potentials.
10)
Allen Steere's
anti-phospholipid antibodies (now known to be caused by activated
Epstein-Barr, as reported by Allen Steere's own Lyme/Lupus reporting team
member, Joe Craft at Yale),
http://www.ncbi.nlm.nih.gov/pubmed/3408508
Full Text:
LYME_AND_LUPUS_STEERE.htm
http://www.ncbi.nlm.nih.gov/pubmed/8410057
11)
the auto-reactive
band 41
[specific to borrelia or not; scientists seem to think an antibody against
flagellin cross reacts with nerve and other tissues, such as skin and intestine
(H. pylori and Campylobacter, producing autoimmune hives, thyroid disease,
GERD...)],
http://www.ncbi.nlm.nih.gov/pubmed/7678336
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC45624/?tool=pubmed
12)
anti-heat shock proteins (a
sign of MS), (and which is a result of anti-flagellar antibodies; Sigal)
http://www.ncbi.nlm.nih.gov/pubmed/7678336
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC45624/?tool=pubmed
13)
anti-gangliosides,
Benach
http://www.ncbi.nlm.nih.gov/pubmed/7558329
14)
GFAp or glial fribrillary acidic protein in the CSF - signs of gliosis
mentioned by Yale's Robert Schoen when he mentioned reasons why we need a
Lyme vaccine,
http://www.annals.org/content/132/8/661.full.pdf+html (gliosis would be a
sign of multiple CNS degenerative diseases),
15)
Gadolinium-Contrast MRI
- performed in monkeys with Lyme but never Lyme-MS Victims which show Lyme is an
active meningitis, and
16)
EMG studies to show whether
or not Lyme victims suffer “hysteria” or the electrification of our hysters.
I think this most of all would be a fun study:
Could we find out via EMG that not-enough-sex really causes Lyme-ALS, or Lyme-MS
or Lyme-Lupus, or Lyme-stroke, or bad knees, or Guillain-Barre?
Who wants to volunteer for that study? And what about men with Low-T? Can that
study be performed? Can we women watch?
We’ll rate their manliness
while this EMG study of their genitals to determine
if not-enough sex is “The Reason for Low T or the Inability of Psychiatrists and
Lawyers to Think like Scientists or Engineers” is performed with our own Checklist.
None of which were
applied by Mark Klempner from 1997 to 2001 when he determined that “there is no
sush thing as Chronic Lyme,” despite being the author of two of the main signs
that it is. The first was Klempner’s MMP-130 only found in the spinal fluid of
chronic Lyme victims and his other two studies from 1992 where he determined and
published the reasons he believed ceftriaxone failed to eradicate all
spirochetes [REF]
