Discrimination of HLA-B5 crossreactive group antigens by human allospecific CTL clones

Human CTL clones discriminating serologically closely related HLA-Bw52, B51, and B35, which belong to HLA-B5 crossreacting group (CREG), were established from peripheral blood lymphocytes by repeated in vitro stimulations. Five HLA-Bw52-specific CTL clones from an individual with HLA-B5 CREG antigens and four CTL clones from another individual with HLA-B51 were generated. The specificity of these CTL clones was ascertained by their lysis of EBV-transformed B cells with HLA-Bw52, but not those with HLA-B51 or B35, and Bw52-transfected Hmy2CIR cells but not HLA-B51 or B35 transfectants. Conversely HLA-B51-specific clones were generated from the HLA-B5 CREG-negative individual, as well as another individual with HLA-Bw52. Their specificity was determined in a similar fashion. Since HLA-B51 differed from HLA-Bw52 only by two amino acid substitutions on the alpha helical region of the alpha 1 domain, these results demonstrated that allospecific CTLs can be produced and discriminate the epitopes formed by the subtle difference in the structure of these HLA class I molecules. Furthermore, three HLA-B35-specific CTL clones were generated from the HLA-B5 CREG-negative individual that discriminated HLA-B35 from HLA-Bw52 and B51. Taken together these results demonstrated that human CTL clones could definitively discriminate the three serologically related HLA-B5 CREG specificities.