Exosomology is very big in industry right now so just
GoogleNews it.
Gram negative
bacteria, TLR2/1 agonist bearers etc, and shed exosomes. This is all *** BIG *** with the NIH. "Exosomes"
or "Vesicles" shed by these bacteria.
OspA-and
similar fungal lipoprotein-containing Blebs
or Vesicles (see arrows in
the below micrograph by Alan Barbour) are shed
lipoproteins, so everyone with "Lyme
disease," also has
LYMErix Disease, because
that is actually the disease
- the chronic exposure to Borrelial
OspA-like blebs resulting in
immunosuppression, tolerance to
mycoplasma in the blood (causes
fatigue),
results in no antibodies being produced,
and the activation of Epstein-Barr/Similars.
http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=7103461
Cadavid and the NIH say these shed
vesicles or blebs go to the brain and
inflame it:
An NIH patent, explaining how Lyme causes
LYMErix-disease:
"The invention relates to novel antigens associated with Borrelia
burgdorferi which are
exported
(or shed) in vivo and whose detection is a means of diagnosing
Lyme disease. The antigens are extracellular membrane vesicles and
other bioproducts including the major extracellular protein antigen.
Another object of the invention is to provide antibodies, monoclonal
and/or polyclonal, labeled and/or unlabeled, that are raised against
the antigens. A further object of the invention is to provide a
method of diagnosing Lyme disease by detecting the antigens in a
biological sample taken from a host using the antibodies in
conventional immunoassay formats. Another object of the invention is
to provide kits, for the diagnosis of Lyme disease, comprising the
antibodies and ancillary reagents. The advantage of the antibodies
used in the invention is that they react with the antigens from
geographically diverse strains of Borrelia burgdorferi, but do not
react with antigens from related Borrelia spirochetes."
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=5,217,872.PN.&OS=PN/5,217,872&RS=PN/5,217,872
2011, THE BLEBS ARE
COVERED IN SLYMERIX>>> "Although we uncovered the existence of at least 10
distinct OM complexes harboring several unique subunits, the complexome is
dominated by the frequent occurrence of a limited diversity of membrane
proteins, most notably P13, outer surface protein (Osp) A, -B, -C, and -D
and Lp6.6."
Characterization of multiprotein complexes of the Borrelia burgdorferi outer
membrane vesicles.
http://www.ncbi.nlm.nih.gov/pubmed/21875077
See where NIH's Martin and Marques say these OspA-ish molecules are
responsible for humoral immunosuppression with chronic brain
inflammation:
http://www.actionlyme.org/151026_OCCAMS-RAZOR.htm
30+ reports:
http://www.ncbi.nlm.nih.gov/pubmed/?term=Borrelia+and+vesicles
This one is by Dave Dorward (Rocky Mountain Bioweapons Labs, NIH), that is
what started it all and Willy Burgdorfer mentioned it in several
conferences. Particularly important, he says this only occurs in gram
negative bacteria (once again the Primers Shell Game report is important):
1990, DNA Is Packaged within Membrane-Derived Vesicles of
GramNegativebut Not Gram-Positive Bacteria
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC184538/pdf/aem00087-0464.pdf
1993, BINGO >>>
Specific adherence of Borrelia burgdorferi extracellular vesicles to human
endothelial cells in culture.
Borrelia burgdorferi
produces extracellular vesicles which contain some of the outer surface
proteins of the bacterium (e.g., OspA and OspB). Borrelial vesicles,
isolated by differential centrifugation and filtration, were tested for the
ability to bind to cultured human umbilical vein endothelial (HUVE) cells in
culture. The recently described lipoprotein OspD was expressed on vesicles.
Vesicles exhibited differential expression of OspB and OspD in a
relationship with passage number and medium serum supplement type,
respectively. Qualitative immunoblotting analyses demonstrated
dose-dependent, passage number-dependent adsorption of vesicles by HUVE
cells. This adsorption was demonstrated to be dependent upon a borrelial
component of the vesicle and not due to the presence of minor contamination
with intact spirochetes. Quantitative experiments examining inhibition of B.
burgdorferi-HUVE association as a function of prior vesicle-HUVE association
demonstrated dependence upon (i) a borrelial component(s) in the vesicle,
(ii) low passage number, and (iii) vesicle protein concentration. However,
vesicle pretreatment of the HUVE cell monolayer was not requisite for this
inhibition. Vesicles from highly passaged borrelias were noninhibitory for
B. burgdorferi-HUVE cell association, regardless of the serum used to
supplement the medium. The use of vesicles as a tool for studying B.
burgdorferi pathogenesis and/or physiology is proposed.
http://www.ncbi.nlm.nih.gov/pubmed/8359911
1993, NIH RML, Garon
HERE IS THE NIH TALKING ABOUT IT SOME MORE IN 1993:
Specific and nonspecific responses of murine B cells to membrane blebs of
Borrelia burgdorferi.
Lymphocyte blastogenesis
assays and immunoblotting were used to investigate and compare murine B-cell
responses to preparations of extracellular membrane blebs (BAg) and
spirochetes (Ag) of Borrelia burgdorferi. Immunoblotting BAg, Ag, and medium
control preparations with serum from naive and infected C57BL/10 mice
revealed that BAg and Ag had similar specific reactivity profiles except
that major antigens of 83, 60, and 41 kDa were detected in Ag but not in BAg.
It was determined that 1 microgram (dry weight) of Ag contained 0.0051 and
0.0063 microgram of outer surface proteins A (OspA) and OspB, respectively,
whereas 1 microgram (dry weight) of BAg contained 0.0024 microgram of OspA
and 0.0015 microgram of OspB. *** Both BAg and Ag caused blastogenesis in
cultures of spleen cells from both groups of mice, but BAg-stimulated
lymphocytes exhibited significantly greater (P < or = 0.05) blastogenesis
after 2 or 6 days of culture than did lymphocytes stimulated by Ag or medium
control.*** Flow cytometry and antibody capture enzyme-linked immunosorbent
assays identified responding lymphocytes as B cells which secreted
polyclonal immunoglobulin M (IgM) but not IgG or IgA. Treatment of BAg and
lipopolysaccharide controls with polymyxin B resulted in as much as 20.7 and
54.3% mean decreases in blastogenesis, respectively. *** Fractionation of
BAg or Ag by ultracentrifugation before culture with spleen cells from naive
mice indicated that B-cell blastogenesis was probably associated with
spirochetal membranes.*** The results of this study demonstrate that
specific humoral responses are directed towards extracellular membrane blebs
which lack the 83-, 60-, and 41-kDa antigens of intact spirochetes and that
blebs also possess significant nonspecific mitogenic activity for murine B
cells. This activity was not due entirely to typical lipopolysaccharide or
OspA and OspB lipoproteins.
http://www.ncbi.nlm.nih.gov/pubmed/8454350
The following reports are
citing the article above ^^^ by the NIH about how the blebs / vesicles /
exosomes cause disease:
http://www.ncbi.nlm.nih.gov/pubmed?linkname=pubmed_pubmed_citedin&from_uid=8454350
2010, Duke
Virulence and immunomodulatory roles of bacterial outer membrane vesicles.
Outer membrane (OM)
vesicles are ubiquitously produced by Gram-negative bacteria during all
stages of bacterial growth. OM vesicles are naturally secreted by both
pathogenic and nonpathogenic bacteria. Strong experimental evidence exists
to categorize OM vesicle production as a type of Gram-negative bacterial
virulence factor. A growing body of data demonstrates an association of
active virulence factors and toxins with vesicles, suggesting that they play
a role in pathogenesis. One of the most popular and best-studied pathogenic
functions for membrane vesicles is to serve as natural vehicles for the
intercellular transport of virulence factors and other materials directly
into host cells. The production of OM vesicles has been identified as an
independent bacterial stress response pathway that is activated when
bacteria encounter environmental stress, such as what might be experienced
during the colonization of host tissues. Their detection in infected human
tissues reinforces this theory. Various other virulence factors are also
associated with OM vesicles, including adhesins and degradative enzymes. As
a result, OM vesicles are heavily laden with pathogen-associated molecular
patterns (PAMPs), virulence factors, and other OM components that can impact
the course of infection by having toxigenic effects or by the activation of
the innate immune response. However, infected hosts can also benefit from OM
vesicle production by stimulating their ability to mount an effective
defense. Vesicles display antigens and can elicit potent inflammatory and
immune responses. In sum, OM vesicles are likely to play a significant role
in the virulence of Gram-negative bacterial pathogens.
http://www.ncbi.nlm.nih.gov/pubmed/20197500
2011, Bartold, SPIROCHETES, LIVING IN YOUR LYMPH NODES (WITH THEIR PARTNERs,
EBV ET AL) WRECKING YOUR IMMUNE SYSTEM. Once again, location being the Force
Multiplier . And whoever is not paying attention will be in jail for a week;
I WILL BE WATCHING YOU....
Lymphoadenopathy during lyme borreliosis is caused by spirochete
migration-induced specific B cell activation.
Lymphadenopathy is a
hallmark of acute infection with Borrelia burgdorferi, a tick-borne
spirochete and causative agent of Lyme borreliosis, but the underlying
causes and the functional consequences of this lymph node enlargement have
not been revealed. The present study demonstrates that extracellular, live
spirochetes accumulate in the cortical areas of lymph nodes following
infection of mice with either host-adapted, or tick-borne B. burgdorferi and
that they, but not inactivated spirochetes, drive the lymphadenopathy. The
ensuing lymph node response is characterized by strong, rapid
extrafollicular B cell proliferation and differentiation to plasma cells, as
assessed by immunohistochemistry, flow cytometry and ELISPOT analysis, while
germinal center reactions were not consistently observed. The
extrafollicular nature of this B cell response and its strongly IgM-skewed
isotype profile bear the hallmarks of a T-independent response. The induced
B cell response does appear, however, to be largely antigen-specific. Use of
a cocktail of recombinant, in vivo-expressed B. burgdorferi-antigens
revealed the robust induction of borrelia-specific antibody-secreting cells
by ELISPOT. Furthermore, nearly a quarter of hybridomas generated from
regional lymph nodes during acute infection showed reactivity against a
small number of recombinant Borrelia-antigens. Finally, neither the quality
nor the magnitude of the B cell responses was altered in mice lacking the
Toll-like receptor adaptor molecule MyD88. Together, these findings suggest
a novel evasion strategy for B. burgdorferi: subversion of the quality of a
strongly induced, potentially protective borrelia-specific antibody response
via B. burdorferi's accumulation in lymph nodes.
http://www.ncbi.nlm.nih.gov/pubmed/21637808
NIH SAYS...
1991, Dorward, Garon:
Immune capture and detection of Borrelia burgdorferi antigens in urine,
blood, or tissues from infected ticks, mice, dogs, and humans.
"The
results indicated that B. burgdorferi appears to shed surface antigens which
are readily detectable in urine, blood, and several organs from infected
hosts."
http://www.ncbi.nlm.nih.gov/pubmed/1864935
2010, - sounds like Barbour's explanation in 1996, re the role blebbing
(which we now know of as exosomes or outer membrane vesicles shed by these
bugs):
B cell activation by outer membrane vesicles--a novel virulence mechanism.
"DNA
associated with OMV [LIKE SPIROCHETAL BLEBS/VESICLES] induced full B cell
activation by signaling through TLR9 [HANDLES EBV, DNA]. Importantly, this
concept was verified in vivo, as OMV equipped with MID and DNA were found in
a 9-year old patient suffering from Moraxella sinusitis. In conclusion,
Moraxella avoid direct interaction with host B cells by redirecting the
adaptive humoral immune response using its superantigen-bearing OMV as
decoys."
http://www.ncbi.nlm.nih.gov/pubmed/20090836
2012,
Seattle Wash,
Membrane Vesicle Release in Bacteria, Eukaryotes, and Archaea: a
Conserved yet Underappreciated Aspect of Microbial Life
"The
production of spherical, membranous vesicles from microbial cell surfaces is
conserved among organisms from all three branches of the tree of life,
spanning both prokaryotes and eukaryotes: Gram-negative and Gram-positive
bacteria (16, 47, 58, 73), archaea (17, 18), fungi (3, 65–67), and parasites
(82, 83). For consistency in this review, we will refer to bacterial and
archaeal structures as membrane vesicles (MVs) and fungal and parasitic
vesicles as either exosomes or shedding microvesicles (two distinct
populations referred to collectively as microvesicles [55]). The microscopic
observation of microbial MVs spans more than 50 years, and numerous
functions have been attributed to these extracellular vesicles by many
investigators. The release of vesicles provides flexibility to respond to
environmental cues, secrete components destined for the cell surface,
virulence factors, and antigens, and interact with the host in the case of
pathogens. Because MV release is conserved across many organisms,
MV-mediated functions are likely to be critical to microbial life.
"Both bacterial MVs and archaeal MVs are derived from the cell surface (Fig.
1A and and2A).2A). Early observation of Gram-negative bacterial MVs revealed
the release of an antigenic complex of lipopolysaccharide (LPS) and
lipoprotein into the surrounding medium following amino acid deprivation of
an Escherichia coli lysine auxotroph (40), which was initially proposed to
be derived from the LPS-containing outer membrane (OM) of the bacteria (29).
Since these early investigations, many groups have developed methodologies
to isolate and analyze bacterial MVs. Although reconciling these differences
in experimental design often makes it difficult to draw generalized
conclusions, it is well accepted that Gram-negative bacterial MVs range from
10 to 300 nm in diameter and contain OM and periplasmic constituents,
including proteins, lipoproteins, phospholipids, and LPS (43, 58). The
contents of the inner membrane (IM) and cytoplasm were generally thought to
be excluded from MVs, although recent analyses of the bacterial MV proteome
suggest that some proteins typically annotated as having cytoplasmic
localization consistently appear in MVs (15, 45, 92, 94). In addition to
bacterial membrane proteins, toxins and signaling molecules can be
incorporated into the membrane or lumen of the MV; MV release then serves as
a secretion mechanism (42, 54, 95). Although Gram-negative bacterial MVs
have been most rigorously studied, recent observation of Gram-positive MV
release has demonstrated that this is a function more widely conserved
across all bacteria. MVs derived from Gram-positive bacteria, such as
Bacillus spp., are similarly sized (50 to 150 nm in diameter [47, 73]) and
are rich in membrane lipids as well as toxins (including the anthrax
toxin)."
....
"TLR ligands in bacterial MVs (including LPS and ***lipoproteins***)
stimulate maturation of and cytokine release by macrophages and dendritic
cells (reference 2 and unpublished observations) and likely contribute to
the pathogenesis of inflammatory infections such as neisserial sepsis (8,
9). Likewise, MVs carrying peptidoglycan, the ligand for the cytosolic
innate immune receptor NOD1, are able to initiate NOD1 signaling and
downstream NF-κB-dependent inflammatory responses (35, 36). Furthermore,
infected antigen-presenting cells may no longer be capable of properly
processing and presenting antigens, potentially inhibiting initiation of
adaptive immune responses. MVs released by pathogenic organisms are
nonviable and contain antigens recognized by the adaptive immune system (2,
4). These surface organelles may therefore represent an important source of
antigen in vivo (Fig. 4E)."
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3370574
1995,
Borrelia burgdorferi vesicle production occurs via a mechanism
independent of immunoglobulin M involvement.
"Borrelia
burgdorferi produces extracellular vesicles containing various borrelial
protein antigens when propagated in vitro in culture media. *** Commonly
observed components of borrelial vesicle preparations are borrelial surface
antigens [LIKE OSPA], bovine serum albumin, and the heavy chains of rabbit
immunoglobulin G and immunoglobulin M. This study employed
ultracentrifugation to harvest borrelial vesicles and analyzed these
preparations by sodium dodecyl sulfate-polyacrylamide gel electrophoresis
and Western immunoblotting. We demonstrated that the rabbit mu heavy-chain
band observed was devoid of OspA or at most levels below those detectable by
immunoblot. We also demonstrated the recovery of borrelial vesicles at
relative centrifugal forces as low as 25,000 x g, compared with the force of
> 200,000 x g normally employed. Further, the mu heavy-chain band was
recovered from uninoculated growth media processed at 25,000 x g, suggesting
that it behaves as a particle rather than as a soluble molecule under these
conditions. Lastly, vesicles were demonstrated to be present in preparations
harvested from growth media supplemented with fetal calf serum, suggesting
that vesicle production by B. burgdorferi can occur in the absence of
immunoglobulins."
http://www.ncbi.nlm.nih.gov/pubmed/7591146
1995, SUNY, Coyle, Benach, Schutzer, SUNY'S BLEB-CAPTURE METHOD:
Detection of Borrelia burgdorferi-specific antigen in antibody-negative
cerebrospinal fluid in neurologic Lyme disease.
"B
burgdorferi antigen can be detected in CSF that is otherwise normal by
conventional methodology, and can be present without positive CSF antibody.
Since CSF antigen implies intrathecal seeding of the infection, the
diagnosis of neurologic infection by B burgdorferi should not be excluded
solely on the basis of normal routine CSF or negative CSF antibody
analyses."
http://www.ncbi.nlm.nih.gov/pubmed/7501150
IL-10 is an immune-suppressor cytokine released also in the presence of OspA
fungo-exosomes or blebs or just plain OspA:
Exosomes released from Mycoplasma infected tumor cells activate
inhibitory B cells.
Mycoplasmas
cause numerous human diseases and are common opportunistic pathogens in
cancer patients and immunocompromised individuals. Mycoplasma infection
elicits various host immune responses. Here we demonstrate that mycoplasma-infected
tumor cells release exosomes (myco+ exosomes) that specifically activate
splenic B cells and induce splenocytes cytokine production. Induction of
cytokines, including the proinflammatory IFN-γ and the anti-inflammatory
IL-10, was largely dependent on the presence of B cells. B cells were the
major IL-10 producers. In splenocytes from B cell deficient μMT mice,
induction of IFN-γ+ T cells by myco+ exosomes was greatly increased compared
with wild type splenocytes. In addition, anti-CD3-stimulated T cell
proliferation was greatly inhibited in the presence of myco+ exosome-treated
B cells. Also, anti-CD3-stimulated T cell signaling was impaired by myco+
exosome treatment. Proteomic analysis identified mycoplasma proteins in
exosomes that potentially contribute to the effects. Our results demonstrate
that mycoplasma-infected tumor cells release exosomes carrying mycoplasma
components that preferentially activate B cells, which in turn, are able to
inhibit T cell activity. These results suggest that mycoplasmas infecting
tumor cells can exploit the exosome pathway to disseminate their own
components and modulate the activity of immune cells, in particular,
activate B cells with inhibitory activity.
http://www.ncbi.nlm.nih.gov/pubmed/22558358
