| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
*
Institute of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106;
Department of Neurology, University of Tübingen, Tübingen, Germany;
InterMune, Brisbane, CA 94010;
Custom Computer Software, Foster City, CA 94404; and
¶ Department of Neurology, University of Texas-Southwestern Medical Center, Dallas, TX 75235
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
MHC molecules are highly polymorphic and have unique binding motifs. Therefore, results on determinant usage obtained in conventional animal models cannot necessarily be extrapolated to Ags presented in the context of human MHC molecules (14). To overcome these limitations, we have applied MHC binding algorithms to predict MOG peptide binding to HLA-DR4 (DRB1*0401), and subsequently probed HLA-DR4 (DRB1*0401) transgenic mice immunized with rhMOG for T cell reactivity to overlapping MOG peptides by computer-assisted cytokine ELISPOT assays at single cell resolution. The data showed that MOG peptide (p)97–108 (core 99–107, FFRDHSYQE) was the immunodominant HLA-DRB1*0401-restricted T cell epitope. This region had a high in vitro binding affinity for HLA-DR4, showed complete sequence homology among mice, rats, and humans, and induced severe experimental autoimmune encephalomyelitis (EAE) in the HLA-DR4 transgenic mice. Interestingly, the same peptide was presented by human HLA-DR4 homozygous B cells, suggesting a role for this epitope in the pathogenesis of MS.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
HLA-DR4 (DRB1*0401) transgenic mice were generated as described previously (15) and bred at Case Western Reserve University (Cleveland, OH) under special pathogen-free conditions. Female transgenic mice were injected at 6–10 wk of age with the Ag in CFA. Pertussis toxin (List Biological Laboratories, Campbell, CA) was added to the immunization regimen as indicated. Recombinant MOG was prepared as described subsequently. Overlapping MOG peptides were obtained from Princeton Biomolecules (Langhorne, PA). CFA was prepared by mixing IFA (Life Technologies, Grand Island, NY) with Mycobacterium tuberculosis H37RA at 5 mg/ml (Difco, Detroit, Michigan). Ags were mixed with the adjuvant to yield a 2-mg/ml emulsion, of which 50 µl was injected s.c. as specified.
Cell separations
Single cell suspensions were prepared from HLA-DR4 lymph node
cells. CD4+ or CD8+ T cells
were obtained by passing the cells through a murine
CD8+ or CD4+ T cell
enrichment column (R&D Systems, Minneapolis, MN) following the
manufacturer’s suggested protocol. FACS analysis showed >95%
enrichment for either CD4+ or
CD8+ T cells. Irradiated syngeneic spleen cells
or EBV-transfected HLA-DR4 (DRB1*0401) homozygous B cells
(16) were added at 1 x 105
cells per well as indicated in Fig. 5
.
|
Cytokine ELISPOT assays were performed as described previously
(17). Briefly, ELISPOT plates (ImmunoSpot; Cellular
Technology, Cleveland, OH) were coated overnight with IFN-
(R46A2, 4
µg/ml)- or IL-5 (TRFK5, 5 µg/ml)-specific capture Ab diluted in 1x
PBS. The plates were blocked with 1% BSA in PBS for 1 h at room
temperature, then washed four times with PBS. Cells from draining lymph
nodes were plated at 5 x 105 cells/well
alone or with MOG peptides (7 µM) in HL-1 medium supplemented with
1% L-glutamine and cultured for 24 h for IFN- and
48 h for IL-5. Subsequently, the cells were removed by washing
four times with PBS then four times with PBS/Tween, and the
biotinylated detection Ab XMG1.2-biotin (2 µg/ml) for IFN- or
TRFK4-biotin (2 µg/ml) for IL-5 were added and incubated overnight.
The plate-bound second Ab was then visualized by adding
streptavidin-alkaline phosphatase (DAKO, Carpenteria, CA) and nitroblue
tetrazolium (Bio-Rad, Hercules, CA)/5-bromo-4-chloro-3-indolyl
phosphate substrate (Sigma-Aldrich, St. Louis, MO). Image
analysis of ELISPOT assays was performed on a Series 1 ImmunoSpot Image
Analyzer (Cellular Technology) as described previously (17, 18). In brief, digitized images of individual wells of the
ELISPOT plates were analyzed for cytokine spots based on the comparison
of experimental wells (containing T cells and APC with Ag) and control
wells (T cells and APC; no Ag). After separation of spots that touched
or partially overlapped, nonspecific noise was gated out by applying
spot size and circularity analysis as additional criteria. Spots that
fell within the accepted criteria were highlighted and counted.
Stimulation index was calculated by dividing the number of cytokine
spots detected in wells pulsed with relevant Ag by the number of
cytokine spots in wells without Ag (medium only). The spot number in
unimmunized or control mice (irrelevant Ag) was in the same range as
the medium controls (not shown). Statistical analysis was performed
with the paired t test or the Mann-Whitney rank sum test
using SigmaStat software (SPSS, Chicago, IL).
Evaluation of clinical disease
Mice were monitored daily for 30 days and on alternate days thereafter. A mean clinical score was assigned for each group using the following scale (19): 0, no abnormality; 1, limp tail; 2, moderate hind limb weakness; 3, complete hind limb paralysis; 4, quadriplegia or premoribund state; 5, death.
Histopathology
At the time of the experiment the brain and spinal cord of the mice were removed and either preserved in 10% formalin or snap-frozen in 2-methyl-butane. Five-micrometer slices of the CNS tissue were prepared and stained with H&E. The tissue was then examined by light microscopy in a blinded fashion by a neuropathologist and evaluated for the extent of inflammation and graded as follows: -, no inflammation; +/-, a few mononuclear cells; +, organization of inflammatory infiltrates around positive vessels; ++, extensive perivascular cuffing with extension into the subarachnoid space; and +++, extensive perivascular cuffing with increasing subarachnoid inflammation (19, 20). Immunofluorescence staining of the brain tissue was performed as described (21). In brief, 5-µm sections of snap-frozen brain tissue were fixed with 4% paraformaldehyde and probed with FITC- or PE-conjugated anti-mouse CD3, CD4, CD8, CD19, MAC-1, and F4/80 Abs (BD PharMingen, San Diego, CA). Images of Ab-labeled tissue sections were captured using a Leica fluorescence microscope equipped with a charge-coupled device camera and image analysis software. Sections were analyzed in a blinded fashion.
Prediction of MOG sequences with high affinity for HLA-DRB1*0401
To identify potential core peptides with predicted high binding affinity for HLA-DRB1*0401, the MOG amino acid sequence was analyzed with two published HLA-DR4 binding algorithms (22, 23). Briefly, a computer program was written that parsed the MOG sequence into successive 9-mers, each beginning one amino acid after the start of the previous 9-mer. The contribution toward binding of each amino acid was summed based on the matrix published by Hammer et al. (22), yielding a peptide score. Higher peptide scores indicate relatively higher affinity binding to HLA-DRB1*0401. Several immunodominant peptides documented in the literature yield relative affinity scores in the range of 4–6 (22). This hierarchy of predicted high affinity peptides was compared with predictions based on the published algorithm of Marshall et al. (23), which calculates an IC50 concentration rather than a relative score. The Marshall et al. (23) algorithm was based on the effects of single amino acid substitutions within the 11-residue core of a 13-mer polyalanine peptide (AAYAAAKAAAAAA) that binds promiscuously to most HLA-DR alleles. The contributions of each of the 11 amino acid residues were multiplied consecutively, and the sum was multiplied by the IC50 of the parent polyalanine peptide (14.7 nM) to derive a predicted IC50 for the sequence. Lower IC50 values indicate higher predicted affinity of a peptide containing the sequence. To compare the affinity of an amino acid sequence predicted by the two algorithms, 9-mer sequences with the anchor residue (F, I, L, M, V, W, or Y) at position 1 were first evaluated by the Hammer et al. (22) algorithm. Each 9-mer, along with the N- and C-flanking amino acids, were then evaluated by the Marshall et al. (23) algorithm, providing an 11-mer peptide with the anchor residue (F, I, L, M, V, W, or Y) at the second position as required by this algorithm.
In vitro peptide binding studies
In vitro binding competition assays were performed as described previously (24). In brief, biotinylated tracer peptides were used in a direct binding assay to establish optimal binding conditions for the purified HLA-DR4 (DRB1*0401) molecules. Relative affinities of MOG peptides for the HLA-DR4 molecules were assessed by an inhibition ELISA based on a dissociation-enhanced lanthanide fluoro-immunoassay (DELFIA; Wallac, Turku, Finland). In the inhibition assay, HLA-DR4 molecules (10 nM) were incubated with fixed amounts of the tracer peptide (class II-associated invariant chain peptide (CLIP)97–120) in the presence of various concentrations (10-fold dilutions between 1 nM and 100 µM) of the unlabeled MOG peptides. The concentration yielding 50% inhibition of binding of the tracer peptide (IC50) was measured by plotting the percentage of inhibition vs the concentration of MOG peptide. Peptides were tested in three independent experiments.
Cloning and bacterial expression of human MOG
The cDNA encoding amino acids 23–247 of human MOG was obtained by reverse transcription of total RNA from the human glioma cell line 45/8. RNA was prepared with TRIzol Reagent (Life Technologies) and first-strand cDNA was synthesized using Moloney murine leukemia virus reverse transcriptase, RNase H Minus (Promega, Madison, Wisconsin), and the gene-specific primer R785. The amplified PCR product was separated by agarose gel electrophoresis on a 1.2% gel and purified with a QIAEXII kit (Qiagen, Hilden, Germany). The cDNA fragment was cloned in Bluescript II KS+ vector (Stratagene, La Jolla, California), which was previously treated with SmaI and alkaline phosphatase according to conventional protocols. The nucleotide sequences of the various clones were determined for both strands by using the PRISM Ready Reaction Dye Deoxy Terminator cycle sequencing kit in conjunction with an Applied Biosystems model 310 DNA Sequencer (both from Applied Biosystems, Foster City, CA). After the purification of the resulting PCR product by phenol:chloroform extraction and ethanol precipitation, the fragment was digested with NcoI and BamHI and ligated into the NcoI/BamHI linearized pQE60 vector (Qiagen). To improve the bacterial expression level, it was necessary to replace some of the original, N-terminal MOG codons by PCR mutation. The nucleotide sequences of the resulting clones were confirmed by sequence analysis, and clone mog-1His was used for expression in Escherichia coli. The His-tagged fusion protein representing the extracellular domain of MOG (amino acids 1–125) was purified under denaturing conditions by metal chelate affinity chromatography on Ni-NTA Agarose columns (Qiagen) according to the manufacturer’s guidelines.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
To characterize the complement of MOG peptides that could bind to
the human HLA-DR4 molecule and could therefore play a role in MS, we
screened the MOG protein sequence with peptide binding algorithms which
had been reported previously to predict the binding of peptides to
HLA-DRB1*0401 (22, 23). These algorithms take into account
the properties of the Ag binding site and of the amino acid side chains
of a peptide to predict its binding to MHC molecules (25).
According to the method developed by Hammer et al. (22), a
numeric value is generated for all 9-mer amino acid sequences of a
given protein, and only peptides with binding scores greater than 2 are
expected to bind to HLA-DRB1*0401. The results for the screening of the
extracellular region of the MOG protein are summarized in Table I. Of these, the sequence MOGp99–107
(FFRDHSYQE) was the peptide with the highest binding score (3.9). This
peptide has an aromatic amino acid (phenylalanine) at the anchor
position P1, and an amino acid with a hydroxyl group (serine) at
position P6, consistent with optimal binding to HLA-DRB1*0401
(26, 27, 28). Several other peptides achieved predicted
binding scores above 2, such as MOGp83–91 (LRIRNVRFS), which has an
aliphatic anchor at P1 (leucine) and a valine at position P6 of the
sequence, and was scored at 2.8. These results were confirmed overall
when compared with a second binding algorithm developed by Marshall et
al. (23). However, the predictions conflicted for some of
the peptides. For example, MOGp83–91 had a good binding score of 2.8
with the Hammer method (22) but had an
IC50 of 17.83 with the Marshall algorithm
(23), indicative of low-affinity binding. Furthermore, the
Hammer algorithm (22) predicted several peptides to be
nonbinders for HLA-DRB1*0401, which were, however, estimated to be
high-affinity binders by the Marshall method (Ref. 23 and
data not shown). Hence, the binding algorithms alone were not
sufficient to unambiguously determine the complement of MOG sequences
that could be targeted by T cells in HLA-DR4+ MS
patients.
|
To test whether or not the predicted and/or any of the other MOG
epitopes were generated in vivo and induced T cell immunity, we used a
transgenic mouse line that expressed HLA-DR4
(DRA-I-E
/HLA-DRB1*0401-I-E
) molecules under the mouse MHC class
II promoter (15). These transgenic mice are deficient for
endogenous mouse MHC class II molecules, have no significant deletions
in their T cell repertoire, and have previously been shown to mount
strong T cell responses to various self and foreign Ags (15, 29).
As shown in Fig. 1, HLA-DR4 transgenic
mice were immunized with the extracellular portion of MOG (rhMOG, amino
acids 1–125), and T cell responses were measured in draining lymph
node cells after recall with overlapping 20-mer MOG peptides by
cytokine ELISPOT assay. Vigorous IFN- responses were detected after
recall with the peptides MOGp91–110 and MOGp81–100, and occasionally
with MOGp21–40. Furthermore, the mice mounted a vigorous Ag-specific
IL-2, but not IL-5, recall response to these peptides (data not shown),
consistent with the induction of Th1 immunity. None of the other MOG
peptides induced significant cytokine production. The same MOG
determinant hierarchy was seen when the mice were tested in parallel in
proliferation assays for recall responses to the overlapping MOG
peptides (data not shown). Mice not immunized, or immunized with
irrelevant control Ags (e.g., hen egg white lysozyme, MBP), showed no
responses to any of the MOG peptides (data not shown), demonstrating
the specificity of the T cell response.
|
B). Cell separation for
CD4 and CD8 T cells demonstrated that the observed T cell responses
were mediated by CD4+, but not by
CD8+, T cells (data not shown).
Taken together, the data showed that MOGp99–107 was the immunodominant
HLA-DR4-restricted MOG peptide, and MOGp80–91 and MOGp21–40 were
subdominant determinants. The results correlated well with the binding
predictions (Table I) and demonstrate that these peptides were
processed and presented in vivo after immunization with the rhMOG
protein. Interestingly, not all of the predicted peptides induced T
cell responses in vivo. These epitopes may either not be processed, or
may not bind well enough to HLA-DRB1*0401 to be presented. There may be
other mechanisms preventing T cell responses for these determinants,
such as tolerance at the T cell level.
In vitro binding of the region MOGp99–107 (FFRDHSYQE) to HLA-DR4 (DRB1*0401)
The data on the predicted binding and ex vivo T cell reactivity
suggested that the region MOGp99–107 had a high affinity for the
HLA-DR4 molecule. However, to formally determine the binding affinity
of this region, purified HLA-DRB1*0401 molecules were incubated with a
biotinylated reference peptide (CLIP, 250 nM), and serial dilutions of
the respective overlapping 9-mer peptides (100–0.001 µM) covering
amino acids 91–108 of the MOG sequence were added. As shown in Fig. 2, this region of the MOG sequence indeed
bound with a high affinity to HLA-DR4, as demonstrated by
IC50 values of 4, 0.07, and 7.4 µM for the
peptides MOGp97–105 (TCFFRDHSY), MOGp98–106 (CFFRDHSYQ), and
MOGp99–107 (FFRDHSYQE) respectively. Thus, the in vitro binding data
confirmed the algorithm prediction.
|
Several HLA-DR4 (DRB1*0401)-restricted MOG epitopes, including the
region MOGp91–110 (core 99–107, FFRDHSYQE), induced T cell immunity
in HLA-DR4 (DRB1*0401) transgenic mice. To establish whether or not
these MOG epitopes played a role in the induction of EAE, we immunized
the HLA-DR4 transgenic mice with MOGp97–108
(TCFFRDHSYQEE) and observed the mice for clinical and
histological signs of EAE (Fig. 3A, •).
Interestingly, immunization of the transgenic mice with this peptide
resulted in severe EAE at 8–14 days after immunization. Brain sections
of mice with EAE stained by H&E showed perivascular and periventricular
inflammatory infiltrates consisting of lymphocytes, macrophages, and
occasional neutrophils (Fig. 4, A and B). Immunofluorescence staining of brain
sections with mAbs demonstrated that the infiltrates consisted of
macrophages/microglia (MAC-1+,
F4/80+) and CD4+ T cells
(Fig. 4, C and D). No B cells or
CD8+ T cells were detected (data not shown).
Furthermore, immunization of the transgenic mice with the subdominant
MOG epitopes MOGp21–40 or MOGp80–91 also induced EAE in the
HLA-DR4 transgenic mice (Fig. 3B, 
and 
,
respectively), demonstrating that these determinants were
encephalitogenic.
|
|
Human B cells process and present the immunodominant MOG epitope
To test whether or not human APCs could present the
encephalitogenic MOG peptides, we purified CD4+ T
cells from MOGp97–108-immunized transgenic mice and stimulated them
with Ag and a human HLA-DR4 homozygous B cell line as APCs
(16). As shown in Fig. 5, the human B cells, pulsed either with the MOGp97–108 peptide or with
the whole rhMOG protein, induced vigorous cytokine production by the
MOGp97–108-specific T cells (filled bars). No T cell response was
induced with the irrelevant control peptide, human collagen type II
p261–273. In contrast, hCIIp261–273-specific CD4+ T cells
(gray bars) responded to the hCII peptide, but not to MOG or the MOG
peptide. Furthermore, fixing of the HLA-DR4+
human B cells in 1% paraformaldehyde completely abrogated the
induction of T cell responses when pulsed with the MOG protein, whereas
the fixed cells induced strong T cell responses to the MOGp97–108
(data not shown). Similarly, HLA-DR4- B cells
failed to induce specific T cell responses to either the MOG protein or
the MOG peptide (data not shown).
Taken together, the data show that human APCs processed and presented the immunodominant HLA-DR4-restricted MOG peptide that we have identified in the transgenic mice. Therefore, the data suggest that this MOG peptide could also play a role in the pathogenesis of human MS.
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
). Hence, our results suggest that it is not sufficient to
identify myelin epitopes as targets for the treatment of autoimmune
diseases based on peptide binding predictions or in vitro binding
assays. Confirmation of the relevance of the autoantigenic epitopes by
in vivo studies is required, such as we have demonstrated in the
HLA-DR4 transgenic mice. Theoretically, myelin epitopes need to be presented in the CNS to activate encephalitogenic T cells. That this actually occurs in vivo has recently been shown for MBPp84–102 in the context of HLA-DR2 (38). However, MBP is an abundant Ag in the CNS, whereas MOG comprises only 0.01–0.05% of the myelin mass (39). Therefore, it is not known whether it is processed and presented in the CNS, and by which type of APCs. It is particularly interesting that in our studies, MOGp97–108-reactive T cells induced EAE, suggesting that this peptide was presented in the CNS. Furthermore, the complete sequence homology of this region with human MOG (and >90% homology for the other MOG peptides identified) suggested that this peptide could similarly be presented in the CNS of MS patients and play a role in the pathogenesis of this disease. The significance of MOG for the autoimmune process is supported by the observation that MOG-reactive T cells are readily detectable in MS patients (40, 41), and MOG immunization induced severe EAE in a non-human primate model of MS (42).
The presented data confirm reports by other investigators that Ag
processing and presentation by human and mouse APCs are
sufficiently similar to permit the identification of antigenic epitopes
within foreign- and self-Ags based on the binding properties of the MHC
molecule (43, 44). Furthermore, by using in our studies
transgenic mice that expressed HLA-DRA-I-E/HLA-DRB1*0401-I-E
molecules under the control of the mouse MHC class II
promoter (15), it was ensured that T cell maturation and
Ag recognition in these mice were unaffected. Thus, the TCR repertoire
of the transgenic mice had no significant deletions, and hence there
was enough plasticity of the T cell repertoire to detect the MOG
epitopes presented in the context of HLA-DRB1*0401 (45).
Most importantly, human B cells pulsed with the MOG protein activated
MOGp97–108-reactive T cells (Fig. 5), demonstrating that this peptide
could be naturally processed and presented to high-avidity T cells in
HLA-DR4-expressing MS patients.
If MOG-reactive T cells have a high avidity for their Ag (experiments are under way in our laboratory to formally test for this), then it may be easier to activate the naive T cell precursors, by molecular mimicry, for example. Furthermore, the frequency of MOG-reactive T cells necessary to induce MS may be very low, perhaps even beyond the detection limit of current assays. Subsequently, T cells specific for other myelin Ags (MBP, PLP) or bystander T cells (specific for unrelated Ags) may cross the blood-brain barrier and perpetuate the MOG-induced disease process.
In summary, we have defined for the first time the nature of the MOG T cell epitopes restricted by human HLA-DR4. Our results show that the HLA-DR4 molecule selected for T cell responses to a high-affinity MHC binding peptide. This peptide was encephalitogenic and was also processed and presented by human APCs. Thus, the identified MOG epitopes are potential targets for Ag-specific immunotherapy.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Thomas G. Forsthuber, Institute of Pathology, School of Medicine, Case Western Reserve University, BRB 936, 2109 Adelbert Road, Cleveland, OH 44106-4943. E-mail address: tgf2{at}pop.cwru.edu
3 Abbreviations used in this paper: MS, multiple sclerosis; EAE, experimental autoimmune encephalomyelitis; MOG, myelin oligodendrocyte glycoprotein; PLP, proteolipid protein; MBP, myelin basic protein; rh, recombinant human; CLIP, class II-associated invariant chain peptide; p, peptide.
Received for publication August 8, 2001. Accepted for publication October 11, 2001.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. J. Immunol. 146:3012.[Abstract]
element usage in response to myoglobin. J. Exp. Med. 174:83.This article has been cited by other articles:
|
|
 |
|
  K. Kawamura, K. Yao, J. A. Shukaliak-Quandt, J. Huh, M. Baig, L. Quigley, N. Ito, A. Necker, H. F. McFarland, P. A. Muraro, et al. Different Development of Myelin Basic Protein Agonist- and Antagonist-Specific Human TCR Transgenic T Cells in the Thymus and Periphery J. Immunol., October 15, 2008; 181(8): 5462 - 5472. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kawamura, K. A. McLaughlin, R. Weissert, and T. G. Forsthuber Myelin-Reactive Type B T Cells and T Cells Specific for Low-Affinity MHC-Binding Myelin Peptides Escape Tolerance in HLA-DR Transgenic Mice J. Immunol., September 1, 2008; 181(5): 3202 - 3211. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. L. Greidinger, Y. J. Zang, K. Jaimes, L. Martinez, M. Nassiri, and R. W. Hoffman CD4+ T Cells Target Epitopes Residing within the RNA-Binding Domain of the U1-70-kDa Small Nuclear Ribonucleoprotein Autoantigen and Have Restricted TCR Diversity in an HLA-DR4-Transgenic Murine Model of Mixed Connective Tissue Disease J. Immunol., June 15, 2008; 180(12): 8444 - 8454. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. K. Murthy, Y. Cong, C. Murphey, M. N. Guentzel, T. G. Forsthuber, G. Zhong, and B. P. Arulanandam Chlamydial Protease-Like Activity Factor Induces Protective Immunity against Genital Chlamydial Infection in Transgenic Mice That Express the Human HLA-DR4 Allele Infect. Immun., December 1, 2006; 74(12): 6722 - 6729. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Burster, A. Beck, E. Tolosa, P. Schnorrer, R. Weissert, M. Reich, M. Kraus, H. Kalbacher, H.-U. Haring, E. Weber, et al. Differential Processing of Autoantigens in Lysosomes from Human Monocyte-Derived and Peripheral Blood Dendritic Cells J. Immunol., November 1, 2005; 175(9): 5940 - 5949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. D. Margot, M. L. Ford, and B. D. Evavold Amelioration of Established Experimental Autoimmune Encephalomyelitis by an MHC Anchor-Substituted Variant of Proteolipid Protein 139-151 J. Immunol., March 15, 2005; 174(6): 3352 - 3358. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Hall, M. A. Vickers, E. McLeod, and R. N. Barker Rh autoantigen presentation to helper T cells in chronic lymphocytic leukemia by malignant B cells Blood, March 1, 2005; 105(5): 2007 - 2015. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Bischof, A. Bins, M. Durr, Y. Zevering, A. Melms, and A. M. Kruisbeek A Structurally Available Encephalitogenic Epitope of Myelin Oligodendrocyte Glycoprotein Specifically Induces a Diversified Pathogenic Autoimmune Response J. Immunol., July 1, 2004; 173(1): 600 - 606. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Vandenbark, C. Rich, J. Mooney, A. Zamora, C. Wang, J. Huan, L. Fugger, H. Offner, R. Jones, and G. G. Burrows Recombinant TCR Ligand Induces Tolerance to Myelin Oligodendrocyte Glycoprotein 35-55 Peptide and Reverses Clinical and Histological Signs of Chronic Experimental Autoimmune Encephalomyelitis in HLA-DR2 Transgenic Mice J. Immunol., July 1, 2003; 171(1): 127 - 133. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Khare, M. Rodriguez, and C. S. David HLA class II transgenic mice authenticate restriction of myelin oligodendrocyte glycoprotein-specific immune response implicated in multiple sclerosis pathogenesis Int. Immunol., April 1, 2003; 15(4): 535 - 546. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Kikuchi, T. Fukazawa, M. Niino, I. Yabe, R. Miyagishi, T. Hamada, S. A. Hashimoto, and K. Tashiro HLA-related subpopulations of MS in Japanese with and without oligoclonal IgG bands Neurology, February 25, 2003; 60(4): 647 - 651. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Weissert, J. Kuhle, K. L. de Graaf, W. Wienhold, M. M. Herrmann, C. Muller, T. G. Forsthuber, K.-H. Wiesmuller, and A. Melms High Immunogenicity of Intracellular Myelin Oligodendrocyte Glycoprotein Epitopes J. Immunol., July 1, 2002; 169(1): 548 - 556. [Abstract] [Full Text] [PDF]
|
|
