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B-Crystallin, a Candidate Autoantigen in Multiple Sclerosis1
Division of Immunological and Infectious Diseases, TNO Prevention and Health, Leiden, The Netherlands
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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B-crystallin, a small stress protein that
was identified previously as an immunodominant Ag of CNS myelin in
multiple sclerosis patients. EBV-induced expression of B-crystallin
in B cells leads to HLA-DR-restricted presentation of the protein and
to activation of proinflammatory B-crystallin-specific Th cells.
While B-crystallin is present in EBV-infected human B cells, the
protein is absent from human lymphoid tissues under normal conditions.
This is in sharp contrast to other stress proteins such as heat-shock
protein (hsp)27 and hsp60 that are ubiquitously expressed in these
tissues. In addition, the absence of B-crystallin from lymphoid
tissues in humans is unique as compared with other mammals. All other
species examined, including rodents, sheep, and primates, showed
constitutive expression of B-crystallin in secondary lymphoid
tissues and sometimes even in the thymus. Since constitutive lymphoid
expression most likely results in immunologic tolerance, such a state
of tolerance to B-crystallin can be expected for all of these
species, but not for humans. When taken together, our data provide
evidence for a novel mechanism by which common viral infections can
trigger myelin-directed autoimmunity in a way that is unique for
humans. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The target Ag(s) of the autoimmune response in MS is still subject of
extensive study. In view of the restricted localization of MS lesions
to CNS white matter, it is likely that pathogenic T cells are locally
triggered by one or more components of the myelin sheath. Previously,
we have shown that in myelin from the brains of MS patients the stress
protein B-crystallin serves as an immunodominant autoantigen to
human T cells (15). Especially at the early active stages of
inflammation and myelin breakdown in the CNS during the formation of MS
lesions, the oligodendrocyte/myelin unit contains elevated levels of
B-crystallin (16).
B-Crystallin is a member of the family of small heat-shock proteins
(hsp) (17). Previous studies have indicated that the in vivo expression
of B-crystallin is different from many other stress proteins,
including family members of hsp60, hsp70, and hsp90, as well as small
stress proteins such as hsp27 (18, 19, 20, 21). As a rule, most stress proteins
are expressed ubiquitously in vertebrates, and they exert routine
housekeeping functions during protein biosynthesis and protein
transport across membranes. Stress-induced accumulation of, for example
hsp60 and hsp70, has been documented for a large variety of cell types
and for many different types of stress. In contrast, constitutive
expression of B-crystallin has only been found in a restricted
number of tissues, including the eye lens and cardiac and skeletal
muscle, while low-level expression has been reported for renal
epithelial cells, lung tissue, and CNS glia cells; in most other
tissues, B-crystallin is undetectable (22, 23, 24). Stress-induced
expression of B-crystallin appears to be similarly restricted. In
humans, stress-induced accumulation of B-crystallin has been
documented for only a limited number of disorders, notably
neurodegenerative diseases, neurotropic infections, and some
malignancies (16, 20, 25, 26, 27, 28). Stress-induced accumulation of
B-crystallin is particularly well documented for human and rodent
astrocytes or cell lines derived thereof (29, 30).
In this study, we show that contrary to hsp27 and hsp60,
B-crystallin is absent from healthy human lymphoid tissues,
including spleen, thymus, and PBL. In contrast, healthy lymphoid cells
from several other mammalian species, including rodents, sheep, and
primates, contain readily detectable levels of B-crystallin.
Infection of human peripheral blood B cells with EBV results in the
expression of B-crystallin and its HLA-DR-restricted presentation to
human T cells. The activated T cells are proinflammatory Th1 cells in
that they secrete large amounts of IFN-
, but little, if any, IL-4.
When taken together, these data indicate that virus-induced
presentation of B-crystallin by human B cells is exceptional since
it is likely to occur in a nontolerized background. The resulting T
cell response is cross-reactive with CNS myelin (15) and, therefore,
may well contribute to the development of MS.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Human PBMC from healthy donors were obtained from the Central Laboratory of The Netherlands Red Cross Blood Transfusion Service (Amsterdam, The Netherlands). PBMC from MS patients, diagnosed as having definite MS, were obtained through Dr. C. H. Polman (Department of Neurology, Free University Hospital, Amsterdam, The Netherlands). Cord blood mononuclear cells (CBMC) were kindly supplied by Dr. C. J. P. Boog, and human spleen by Dr. N. M. Lardy (Central Laboratory of The Netherlands Red Cross Blood Transfusion Service). Human thymus was provided by Dr. M. Verschuren (Erasmus University, Rotterdam, The Netherlands). EBV-negative B cell lymphomas of patients diagnosed with monoclonal gammopathy were kindly provided by Dr. J. W. Gratama (The Daniel den Hoed Clinic, Rotterdam, The Netherlands).
Female SJL mice were obtained from Erasmus University. Female BALB/cbyJIco (BALB/c), C57BL/6JIco (C57BL/6J), and 129/SvPasIco (129/Sv) mice were obtained from the Broekman Institute (Someren, The Netherlands). Male Lewis/CrIBr (Lewis) rats were purchased from Charles River (Wilmington, MA)/the Broekman Institute. Female C57BL/6KH mice and male BN/BiRy (Brown Norway) rats were bred in our animal facilities. Peripheral blood of a sheep was a kind gift of veterinary surgeon Dr. P. Vugts (Oud-Gastel, The Netherlands). Peripheral blood samples of various primates were kindly provided by Dr. B.‘t Hart and H. Brok (Biomedical Primate Research Center, Rijswijk, The Netherlands).
Single cell suspensions of thymus, spleen, and cardiac muscle were prepared. PBMC and splenocytes were isolated by density-gradient centrifugation. Cells were washed twice with PBS, frozen in liquid nitrogen, and stored at -70°C until RNA isolation was performed.
Generation of B-crystallin-specific T cell lines
PBMC from either healthy control subjects or MS patients were
cultured in RPMI 1640 (Dutch modification) supplemented with 100 U ·
ml-1 penicillin, 0.1 mg · ml-1
streptomycin, 2 mM L-glutamine, 1 mM sodium pyruvate, 10 mM
HEPES (pH 7.4), and 5% (v/v) pooled human serum at 2 x
105 cells per 100 µl in round-bottom wells at 37°C in
the presence of 5 µg · ml-1 B-crystallin.
B-Crystallin was purified from bovine eye lenses by reversed-phase
HPLC (15). On days 5 and 8, 10% (v/v) Lymphocult-T (Biotest Seralc,
Zaventem, Belgium) was added as a source of growth-promoting cytokines
including IL-2. Ag specificity was evaluated by split-well assays on
day 11 or 12. Specific T cell lines (TCL) were kept in culture by
weekly stimulation with 106 ml-1 irradiated
(30 Gy) autologous PBMC in the presence of 5
µg · ml-1 B-crystallin for at least 7 wk. Bovine
B-crystallin is identical to the human homologue, except for
conservative substitutions at 4 of the 175 amino acid positions.
T cell proliferation assays
Proliferation assays were performed by seeding 5 x
104 T cells with 105 irradiated (30 Gy)
autologous PBMC and varying doses of B-crystallin in 200 µl medium
in round-bottom wells. After 72 h of culture, 20 kBq
[3H]thymidine (Amersham Life Sciences, Arlington Heights,
IL) was added per well, and, after another 16 h, thymidine
incorporation was determined by using a beta plate counter (Canberra
Packard, Meriden, CT). To determine TCL responses using EBV-LCL
as APC, TCL were seeded at 5 x 104/well in the
presence of 5 x 104 irradiated (50 Gy) autologous
EBV-LCL as APC and varying doses of purified B-crystallin. The
effect of anti-HLA Abs on TCL responses was examined by
proliferation assays in the presence of 10 µg · ml-1
B-crystallin. APC were preincubated for 30 min with mAbs B8.11.2
(anti-HLA-DR, IgG2b), SPV-L3 (anti-HLA-DQ, IgG2a), or B9.12.1
(anti-HLA-class I) (31).
Flow cytometry
The expression of various surface Ags on
B-crystallin-specific TCL was analyzed by direct immunofluorescence
dual staining. For this purpose, T cells were incubated with mAbs
directed at CD3, CD4, CD8, ß-TCR, or 
-TCR conjugated with
FITC or phycoerythrin (Becton Dickinson) and analyzed on a FACScan flow
cytofluorometer using Cell Quest software (Becton Dickinson).
Cytokine assays
To examine cytokine release by TCL in response to
B-crystallin, 5 x 105 T cells were seeded with
106 autologous, irradiated PBMC and 5 µg B-crystallin
in 1 ml of culture medium. After 3 days, culture supernatants were
examined by ELISA for the presence of IFN- (Pelikine Compact,
Central Laboratory of The Netherlands Red Cross Blood Transfusion
Service) and IL-4 (32).
RT-PCR
mRNA was isolated using the RNAzol B method (Campro Scientific,
Veenendaal, The Netherlands) and isopropanol precipitation.
Using 1 µg of mRNA as a template, copy DNA was produced using the
Reverse Transcription System (Promega, Madison, WI). For amplification,
cDNA (1 µl) was added to 1 µl 10 mM dNTP mix, 5 µl 10x
Taq polymerase buffer (500 mM KCl, 600
µg · ml-1 BSA, 100 mM Tris-HCl (pH 8)), and 1 unit
Taq DNA polymerase (Life Technologies, Amsterdam, The
Netherlands) in a final volume of 50 µl. Primers and amplification
conditions were as follows: B-crystallin, 5'-AGCTGGTTTGACACTGGACT-3'
and 5'-CAATTCAAGAAAGGGCATC-3', 35 cycles of melting at 94°C,
annealing at 65°C, and extension at 72°C for 30 s each in the
presence of MgCl2 at a final concentration of 3 mM;
ß-actin, 5'-AAGATGACCCAGATCATGTTTGAG-3' and
5'-AGGAGGAGCAATGATCTTGATCTT-3', 35 cycles of melting at 94°C,
annealing at 65°C, and extension at 72°C for 30 s each in the
presence of MgCl2 at a final concentration of 2.25 mM;
Hsp27, 3'-CAAAAGAACACACAGGTGCG-5' and 3'-TCCCTGGATGTCAACCACTT-5',
40 cycles of melting at 94°C, annealing at 60°C, and extension at
72°C for 60 s each in the presence of MgCl2 at a
final concentration of 1.5 mM; Hsp60, 3'-TTGGGCTTCCTGTCACAGTT-5' and
3'-TTCGATGCATTCCAGCCTTG-5', 40 cycles of melting at 94°C,
annealing at 60°C, and extension at 72°C for 60 s each in the
presence of MgCl2 at a final concentration of 2.25 mM.
PCR was conducted in a Perkin-Elmer Cetus GenAmp System 9600
(Perkin-Elmer, Norwalk, CT). PCR products were run on 2% agarose gel
by electrophoresis. The transformed astroglioma cell line U373 that
constitutively expresses B-crystallin was used as a source of
reference mRNA.
To amplify B-crystallin-encoding mRNA from various species,
pan-B-crystallin-specific primers were designed to match all known
sequences. cDNA (3 µl) was added to 1 µl 10 mM dNTP mix, 5 µl
10x Taq polymerase buffer (500 mM KCl, 600
µg · ml-1 BSA, 100 mM Tris-HCl (pH 8), 30 mM
MgCl2), and 1 unit Taq DNA polymerase (Life
Technologies) in a final volume of 50 µl.
Pan-B-crystallin primers were 5'-TGCRGTGACAGCAGGCTTCT-3' and
5'-GAGAGCACCTGTTGGAGTCT-3'. Reaction conditions included premelting at
94°C for 4 min, followed by 40 cycles of melting at 94°C, annealing
at 65°C, and extension at 72°C for 30 s each.
Western blot analysis
Whole cell lysates derived from EBV-LCL or the corresponding
original PBMC were prepared by lysis in 20% (v/v) acetic acid supplied
with 8 M urea. Standard SDS-PAGE analysis was done using an 8–25%
gradient polyacrylamide gel (Pharmacia Biotech, Piscataway, NJ). For
Western blotting, a 1/25 dilution was used of polyclonal rabbit Abs
against human B-crystallin that was isolated from MS-affected brains
(16).
Viral infection of CBMC
CBMC were cultured in RPMI 1640 supplemented with 5% (v/v) pooled human serum at 106 ml-1 in a final volume of 10 ml in 25-cm2 culture flasks. The cells were incubated for 48 h with 2.5 ml marmoset B95-8 cells culture supernatant containing EBV, 25 x 103 ml-1 Tissue Culture Infectious Dose50 HHV-6 (strain U110Z) (33), 5 x 105 plaque-forming units per ml MV (strain Edmonston) (34), 33 hemagglutinating units per ml influenza virus A/HK/68, or 10 µg · ml-1 LPS (Escherichia coli 0127:B8; Difco, Brunschwig Chemie, The Netherlands).
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B-crystallin-specific T cells proliferate in
response to autologous EBV-LCL in the absence of exogenous Ag
Panels of TCL were generated from PBMC by weekly stimulation with
purified B-crystallin in the presence of irradiated, autologous PBMC
as APC. As a rule, B-crystallin-specific TCL were
CD4+CD8-, expressed a TCR-ß, and
were HLA-DR restricted in their response to B-crystallin
(representative data for selected TCL are given in Fig. 1
, A and B). All
TCL were of the Th1 phenotype in that they produced significant amounts
of the proinflammatory cytokine IFN- in response to B-crystallin
and little, if any, IL-4 (Fig. 1C). To date, no qualitative
differences were found between TCL derived from either MS patients or
healthy control subjects.
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B-crystallin-specific TCL showed the
expected dose-dependent response to their target Ag in the presence of
autologous PBMC, they responded vigorously to autologous EBV-LCL in the
absence of any exogenously supplied Ag. Representative data for three
TCL are shown in Fig. 2A. In
some (e.g., line S5L5), but not in all cases, addition of exogenous
B-crystallin resulted in a further enhancement of proliferative
responses in a dose-dependent manner. The proliferative response to
EBV-LCL both in the absence or presence of exogenously added
B-crystallin was HLA-DR restricted in that no responses were
observed to HLA-DR-mismatched allogeneic EBV-LCL (data for S5L5 given
in Fig. 2B) and that responses to autologous EBV-LCL could
be blocked by mAbs to HLA-DR, but not to HLA-DQ (data not shown).
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B-crystallin expression
One explanation for the above data would be HLA-DR-restricted
presentation of endogenously produced B-crystallin by EBV-LCL.
Expression of B-crystallin was examined by amplifying mRNA by the
reverse-transcriptase PCR (RT-PCR) using human B-crystallin-specific
primers. In all cases, readily detectable levels of
B-crystallin-encoding mRNA were present in EBV-LCL, but not in
populations of PBMC before infection and transformation. Fig. 3A shows representative data
obtained for two different donors. In accordance with these data,
Western blot analysis of whole cell lysates revealed detectable levels
of B-crystallin only in EBV-transformed cells and not in untreated
PBMC from the same donors (Fig. 3B). Finally,
B-crystallin expression in EBV-LCL was confirmed by
immunocytochemistry, which revealed cytosolic and nuclear expression of
the protein, while no such staining could be detected in unmanipulated
PBMC (data not shown). The above findings indicate that upon infection
and transformation by EBV, peripheral blood B cells start to synthesize
B-crystallin, leading to its presentation by HLA-DR molecules.
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B-crystallin expression in human lymphocytes results from viral
infection rather than from transformation
Next, we examined whether or not endogenous expression of
B-crystallin in human lymphocytes is a unique consequence of EBV
infection or whether perhaps other viruses or stimuli could have a
similar effect. Human umbilical CBMC, free of prior viral infection,
were exposed to a variety of stimuli, including infectious viruses.
Expression of B-crystallin-encoding mRNA was determined by RT-PCR
following a 48-h culture period. Fig. 4
shows that B-crystallin-encoding mRNA cannot be detected in
unmanipulated cord blood cells nor in cells cultured in the absence of
any stimulus. In vitro infection of cord blood cells with EBV, MV,
HHV-6, or, to a lesser extent, influenza A virus triggered the
production of detectable levels of B-crystallin-specific mRNA within
a 48-h period. B cell-stimulating LPSs failed to produce such an
effect.
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B-crystallin expression in EBV-LCL could
perhaps be associated with B cell transformation rather than with
virus-associated events, we examined a panel of 10 EBV-negative
human B cell lymphomas derived from patients with monoclonal
gammopathies for the presence of B-crystallin-encoding mRNA. In 5 of
10 EBV-negative B cell lymphomas, only trace levels of
B-crystallin-encoding mRNA could be detected at levels much lower
than what can routinely be found in EBV-LCL. In the other five
lymphomas, no specific message was detectable at all (data not shown).
Apparently, accumulation of B-crystallin in lymphocytes is not
uniquely associated with EBV infection, but still is an event of some
selectivity and associated with viral infection rather than with mere
transformation of B cells.
B-Crystallin expression in peripheral lymphoid tissues is
different from other hsp and varies among mammalian species
While the data in Figs. 3
and 4 already indicate that
B-crystallin is absent from healthy human PBL, the expression of
B-crystallin in PBMC was further examined in a larger panel of
subjects and compared with other stress proteins by RT-PCR analysis.
For this comparison, we focused on hsp27 as another small stress
protein and on hsp60 as a representative of large stress proteins. The
data consistently showed that B-crystallin-encoding mRNA is absent
from human PBMC, while readily detectable levels of mRNA-encoding hsp27
and hsp60 were found in all samples. Fig. 5 gives the results for a panel of six
healthy donors.
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B-crystallin
expression to other lymphoid tissues that are relevant to the induction
and maintenance of immunologic tolerance, i.e., thymus and spleen. At
the same time, we compared the expression of B-crystallin in human
tissues with corresponding tissues from several other mammalian
species. For this purpose, PBMC and, when possible, spleen and thymus
were collected from various strains of mice and rats, and sheep and
primates, including chimpanzees, rhesus monkeys, marmosets, aotus,
cynomolgous monkeys, and cotton-top tamarins. To deal with the
(limited) sequence differences in B-crystallin-encoding mRNAs in
these different species, a set of pan-B-crystallin primers was
designed that match all known sequences. As a positive control, cardiac
muscle from as many different species as possible was taken as a source
of constitutively expressed species-specific mRNA. The human
astroglioma cell line U373 that constitutively expresses
B-crystallin was used as a positive control for the amplification of
human B-crystallin-encoding mRNA.
The results of these analyses, shown in Fig. 6, clearly illustrate that in contrast to
the absence of expression in human secondary lymphoid tissues,
B-crystallin-encoding mRNA is readily detectable both in PBMC and in
spleen cells of other mammals (Fig. 6, A and B).
This holds for all rodents examined and for most primates as well. As
an exception, B-crystallin-encoding mRNA was undetectable in SJL
spleen despite expression in PBMC. At present, we have no explanation
for this apparent discrepancy. Amplifications using pan-B-crystallin
primers failed to produce amplicons from chimpanzee as well as from
cotton-top tamarin PBMC. When the amplification was repeated with the
set of human-specific primers, cotton-top tamarin and chimpanzee
PBMC remained negative (data not shown). When cardiac muscle was
available as a control, amplification of species-specific mRNAs using
the pan-B-crystallin primers was always positive (Fig. 6D). In the case of chimpanzees and cotton-top tamarins, no
cardiac muscle was available as a positive control nor information on
the sequence of B-crystallin in these species. Therefore, the
possibility that in these two cases no specific message could be
amplified from PBMC as the result of inadequate primer matching cannot
be excluded.
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C further show that expression of
B-crystallin in the thymus is more restricted. Clearly, detectable
levels of B-crystallin were found in the thymus of only 129/Sv and
C57BL/6KH mice and Lewis rats, while trace amounts were detected in the
BALB/c sample. All other samples analyzed were negative. |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B-crystallin. Contrary to other major
stress proteins, B-crystallin is not expressed constitutively in
human secondary or primary lymphoid tissues, including PBMC, spleen,
and thymus. This lack of expression is a property that humans may only
share with chimpanzees and cotton-top tamarins, but absence of
expression in PBMC of these latter species could not be formally proven
due to the lack of positive control tissue. All other mammals examined
displayed readily detectable constitutive levels of B-crystallin in
secondary lymphoid tissues, and in some cases even in the thymus.
Most likely, constitutive expression of B-crystallin in secondary
(and in some cases primary) lymphoid organs in most mammals results in
a state of natural peripheral (and in some cases central) tolerance for
the protein. In agreement with this notion, earlier reports have
documented murine T cell tolerance for eye lens crystallins, including
B-crystallin (35, 36). Recent data confirm that thymic expression of
autoantigens in rodents results in resistance to experimental
autoimmune disease as can otherwise be induced by these Ags (37).
Conversely, the lack of expression of B-crystallin in human lymphoid
cells may render the human immune system not tolerant to the protein.
Not only would this condition of humans be unique as compared with
other mammals, it would also be quite different from the state of
functional tolerance that must exist for other stress proteins,
including hsp27, hsp60, hsp70, and hsp90, since these are expressed
constitutively at appreciable levels in all human lymphoid tissues.
Against the above background, the appearance of B-crystallin in
virus-infected human B cells and the resulting presentation of the
protein via HLA-DR to T cells may have interesting consequences in
humans. Different from what would happen with other stress proteins and
different from what would happen in most other mammals, generation of a
T cell response to B-crystallin is the likely result. All
B-crystallin-specific T cells that we have isolated to date from
either MS patients or healthy control subjects are classical
CD4+Th1 cells that bear the ß-TCR and produce IFN-
and little, if any, IL-4 in response to Ag. At least in vitro, EBV, MV,
HHV-6, and, to a lesser extent, influenza A virus appear to be able to
switch on expression of B-crystallin in human lymphocytes. Our data
suggest that EBV is particularly effective in this respect.
As was shown previously, B-crystallin is a dominant CNS myelin Ag
for human Th cells in the CNS of MS patients (15). In addition, recent
data show that subdominant or cryptic epitope of B-crystallin is
capable of inducing experimental allergic encephalomyelitis in mice
(N.-M. Thoua, J. M. Van Noort, M. M. Morris, D. Baker, A.
Bose, A. C. Van Sechel, M. J. B. Van Stipdonk, P.
J. Travers, and S. Amor, in preparation). Thus, a virus-generated
memory T cell repertoire specific for B-crystallin that primarily
involves Th1 cells is cross-reactive with CNS myelin and should be
considered as potentially pathogenic. When such a T cell repertoire
would be recruited into the CNS under conditions that also lead to the
local accumulation and presentation of B-crystallin, a pathogenic
inflammatory burst could well be the result. Yet, each of the
contributing factors, including the presence of a virus-induced
peripheral repertoire against B-crystallin, would not necessarily be
pathogenic on their own. Recent data show that recruitment of
lymphocytes and accumulation of B-crystallin in the
oligodendrocyte/myelin are precisely what can be observed in early
active stages of lesional development in MS, while they are not
observed in inactive plaque areas or healthy control tissue (16).
It is tempting to relate the presently observed capacity of EBV to
trigger presentation of B-crystallin in secondary lymphoid cells to
the existing epidemiologic evidence implying EBV infection as a risk
factor in the development of MS (11, 13, 14). Several studies have
shown that 99–100% of MS patients are EBV seropositive (38, 39),
consistent with EBV infection being a necessary (but by no means
sufficient) contributing factor. In addition, evidence has been
reported to indicate that an episode of infectious mononucleosis, which
is characterized by an exaggerated T cell response to EBV-infected B
cells, significantly increases the risk to develop MS later on in life
(11, 12, 14, 40, 41). While evidence for an involvement of EBV in the
development of MS remains only circumstantial, data such as the above
cannot be fully ignored. Our present study provides a novel mechanism
by which the contributions to MS of relatively common viral infections
on one hand and CNS myelin-directed T cell responses on the other may
be reconciled.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. J. M. van Noort, Division of Immunological and Infectious Diseases, TNO Prevention and Health, P.O. Box 2215, 2301 CE Leiden, The Netherlands. E-mail address:
3 Abbreviations used in this paper: MS, multiple sclerosis; CBMC, cord blood mononuclear cell; CNS, central nervous system; EBV-LCL, EBV-transformed lymphoblastoid cell line; HHV-6, human herpes virus-6; hsp, heat-shock protein; MV, measles virus; TCL, T cell line.
Received for publication June 2, 1998. Accepted for publication September 4, 1998.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B-crystallin as candidate autoantigen in multiple sclerosis. Nature 375:798.[Medline]
B-crystallin in glia cells during lesional development in multiple sclerosis. J. Neuroimmunol. 78:143.[Medline]
B-Crystallin is a small heat shock protein. Proc. Natl. Acad. Sci. USA 88:3652.B-crystallin in the absence of stress. J. Cell Biol. 120:639.B-crystallin and hsp27 expression in response to physiological stress. J. Cell. Physiol. 159:41.[Medline]
B-crystallin gene is not restricted to the lens. Mol. Cell. Biol. 9:1083.B-crystallin in non-lenticular tissues. J. Histochem. Cytochem. 38:31.[Abstract]
ß-crystallin in skeletal muscle: purification and localization. J. Biochem. 110:812.B-Crystallin expression in non-lenticular tissues and selective presence in ubiquitinated inclusion bodies in human disease. J. Pathol. 166:61.[Medline]
B-Crystallin and 27-kd heat shock protein are regulated by stress conditions in the central nervous system and accumulate in Rosenthal fibers. Am. J. Pathol. 143:487.[Abstract]
B-crystallin and HSP27 in Alexander Disease. Am. J. Pathol. 143:1743.[Abstract]
B-crystallin in epithelial tumors: a useful tumor marker?. J. Pathol. 174:209.[Medline]
B-Crystallin accumulation in human astroglioma cell line U373 MG is stress-dependent and phosphorylation-independent. J. Biochem. 116:973.B-crystallin in C6 glioma cells is induced by agents that promote the disassembly of microtubules. J. Biol. Chem. 271:26989.-crystallin in knockout mice. Invest. Ophthalmol. Vis. Sci. 38:S491. (Abstr.).
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 A. Ascherio, K. L. Munger, E. T. Lennette, D. Spiegelman, M. A. Hernan, M. J. Olek, S. E. Hankinson, and D. J. Hunter Epstein-Barr Virus Antibodies and Risk of Multiple Sclerosis: A Prospective Study JAMA, December 26, 2001; 286(24): 3083 - 3088. [Abstract] [Full Text] [PDF]
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 K.-P. Wandinger, W. Jabs, A. Siekhaus, S. Bubel, P. Trillenberg, H.-J. Wagner, K. Wessel, H. Kirchner, and H. Hennig Association between clinical disease activity and Epstein-Barr virus reactivation in MS Neurology, July 25, 2000; 55(2): 178 - 184. [Abstract] [Full Text] [PDF]
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 J. J. Bajramovic, A. C. Plomp, A. v. d. Goes, C. Koevoets, J. Newcombe, M. L. Cuzner, and J. M. van Noort Presentation of {alpha}B-Crystallin to T Cells in Active Multiple Sclerosis Lesions: An Early Event Following Inflammatory Demyelination J. Immunol., April 15, 2000; 164(8): 4359 - 4366. [Abstract] [Full Text] [PDF]
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B. L. Banwell and A. G. Engel {alpha}B-Crystallin immunolocalization yields new insights into inclusion body myositis Neurology, March 14, 2000; 54(5): 1033 - 1041. [Abstract] [Full Text] [PDF]
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