Ex vivo detection of myelin basic protein-reactive T cells in multiple sclerosis and controls using specific TCR oligonucleotide probes.
T cell reactivity to candidate myelin autoantigens, such as myelin basic protein (MBP), may play an important role in the pathogenesis of multiple sclerosis (MS). Although MBP-reactive T cells have been found to undergo in vivo activation in patients with MS, their true precursor frequency in MS is unknown as current frequency analysis is commonly based on the T cell functional responses to MBP. In this study, we developed a TCR sequence-based ex vivo detection system using colony hybridization with oligonucleotide probes specific for CDR3 of selected T cell clones for the analysis of true T cell precursor frequency in PBMC. The results revealed that the precursor frequency of five independent T cell clones recognizing the immunodominant MBP(83-99) region was found to be in the range of 1.6 x 10(-4) in total T cells in three HLA-DR2 patients with MS compared to that of 0.25 x 10(-4) in HLA-DR2 healthy individuals. The observed frequency of MBP(83-99)-reactive T cells in MS patients was considerably higher than those measured in parallel by cell culture-based analysis (2.3 x 10(-6)) or by enzyme-linked immunospot assay (3.9 x 10(-5)) in the same peripheral blood mononuclear cell specimens. Furthermore, the study showed that MBP(83-99)-reactive T cells detected ex vivo belonged to CD45RA+, CD25+ and CD95- T cell subsets as evidenced by preferential expression of specific TCR transcripts in these cell fractions.
Hong
Jian
J
Department of Neurology and Baylor-Methodist Multiple Sclerosis Center, Baylor College of Medicine, Houston, USA.|Joint Immunology Laboratory of Health Science Center and Shanghai Institute of Immunology, Shanghai Institutes of Biological Sciences - Shanghai Second Medical University, Shanghai, China.
Zang
Ying C Q
YCQ
Department of Neurology and Baylor-Methodist Multiple Sclerosis Center, Baylor College of Medicine, Houston, USA.|Joint Immunology Laboratory of Health Science Center and Shanghai Institute of Immunology, Shanghai Institutes of Biological Sciences - Shanghai Second Medical University, Shanghai, China.
Li
Sufang
S
Department of Neurology and Baylor-Methodist Multiple Sclerosis Center, Baylor College of Medicine, Houston, USA.
Rivera
Victor M
VM
Department of Neurology and Baylor-Methodist Multiple Sclerosis Center, Baylor College of Medicine, Houston, USA.
Zhang
Jingwu Z
JZ
Department of Neurology and Baylor-Methodist Multiple Sclerosis Center, Baylor College of Medicine, Houston, USA.|Joint Immunology Laboratory of Health Science Center and Shanghai Institute of Immunology, Shanghai Institutes of Biological Sciences - Shanghai Second Medical University, Shanghai, China.|Immunology Division, E-Institutes of Shanghai Universities, Shanghai, China.
D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D001324:Autoantigens / Q000276:immunology*; D001483:Base Sequence; D002478:Cells, Cultured; D002999:Clone Cells; D022801:Complementarity Determining Regions / Q000235:genetics; D005260:Female; D019672:Genes, T-Cell Receptor; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000276:immunology*; D004676:Myelin Basic Protein / Q000276:immunology*; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013234:Stem Cells / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D015703:Antigens, CD; D001324:Autoantigens; D022801:Complementarity Determining Regions; D004676:Myelin Basic Protein; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; C104448:myelin basic protein 83-99