A translatable molecular approach to determining CD8 T-cell epitopes in TMEV infection.
Defining the epitope specificity of CD8+ T cells is an important goal in autoimmune and immune-mediated disease research. We have developed a translational molecular approach to determine the epitope specificity of CD8+ T cells using the Theiler's murine encephalomyelitis virus (TMEV) model of multiple sclerosis (MS). TMEV-specific CD8+ T cells were isolated from brains and spleens of 7-day TMEV-infected C57BL/6J mice and stimulated by Cos-7 cells that were co-transfected with expression vectors encoding the D(b) class I molecule along with overlapping segments of the TMEV genome. Both brain-infiltrating and spleen-derived CD8+ T cells expressed IFN-gamma when Cos-7 cells were co-transfected with D(b) class I molecule and the TMEV genomic segment that encoded the immunodominant TMEV epitope. This demonstrated that peripheral and brain-infiltrating CD8+ T-cell responses were focused on peptide epitope(s) encoded by the same region of the TMEV genome. We propose that a similar molecular approach could also be used to determine the antigen specificity of suppressor CD8 T cells by the measurement of transforming growth factor-beta (TGF-beta) production. In addition, with a randomly generated library and peripheral blood or isolated CSF CD8+ T cells, this would be an effective method of predicting the epitope specificity of CD8+ T cells in human inflammatory CNS diseases, in animal models of MS or other organ-specific inflammatory diseases with a protective or pathogenic role of CD8 T cells.
McDole
Jeremiah
J
Department of Neurology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Suidan
Georgette
G
Boespflug
Erin
E
Cheng
Jie
J
Chen
Yi
Y
Pirko
Istvan
I
Johnson
Aaron
A
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D001921:Brain / Q000276:immunology* / Q000473:pathology / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000473:pathology; D019556:COS Cells; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D016679:Genome, Viral / Q000235:genetics / Q000276:immunology; D006801:Humans; D008297:Male; D051379:Mice; D009103:Multiple Sclerosis / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D011051:Poliomyelitis / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D017945:Theilovirus / Q000235:genetics / Q000276:immunology*
D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D010455:Peptides