Selection of encephalitogenic rat T-lymphocyte clones recognizing an immunodominant epitope on myelin basic protein.
Using the soft agar-cloning technique, we isolated 13 T-cell clones from guinea pig basic protein (GP-BP)-specific T-cell lines derived from Lewis rats. The clonal frequency was approximately 2.5 x 10(-5). Each of these clones had a similar but not identical pattern of response to a battery of synthetic peptides representing overlapping epitopes in the encephalitogenic region for Lewis rats (69-89 sequence). All clones responded to the minimal encephalitogenic sequence (residues 72-84) restricted by I-A but not I-E molecules, and all transferred clinical experimental autoimmune encephalomyelitis (EAE) and delayed-type hypersensitivity (DTH) reaction to naive rats. Phenotypically, the clones were W3/13+ (total T), W3/25+ (T helper), and OX-22+ (DTH associated). This report demonstrates for the first time the applicability of the soft agar-cloning technique for obtaining encephalitogenic T-cell clones. The exclusive recovery of 72-84-specific T-cell clones after only two rounds of stimulation with GP-BP indicates the immunodominance of this epitope and the power of the line selection technique for obtaining encephalitogenic T-cell specificities.
Chou
Y K
YK
Neuroimmunology Research Laboratory, Veterans Administration Medical Center, Portland, Oregon 97201.
Vandenbark
A A
AA
Jones
R E
RE
Hashim
G
G
Offner
H
H
D000362:Agar; D000818:Animals; D002478:Cells, Cultured; D002999:Clone Cells; D046508:Culture Techniques / Q000379:methods; D004261:DNA Replication; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology; D000939:Epitopes / Q000032:analysis*; D006968:Hypersensitivity, Delayed; D007116:Immunization, Passive; D008213:Lymphocyte Activation; D004676:Myelin Basic Protein / Q000276:immunology*; D051381:Rats; D011917:Rats, Inbred Lew; D013601:T-Lymphocytes / Q000276:immunology* / Q000528:radiation effects
D000939:Epitopes; D004676:Myelin Basic Protein; D000362:Agar