Generation of cytotoxicity with various MHC class I restrictions against autologous LCL by stimulating the lymphocytes with autologous and/or allogeneic LCLs sharing HLA alleles with the responder

Abstract

We exposed human blood lymphocytes to autologous and to allogeneic lymphoblastoid lines (LCLs), each alone or in combination, and analyzed the MHC Class I restriction pattern of the generated auto-LCL reactive cytotoxicity. In the cultures of two EBV-seropositive, HLA All-positive individuals the majority of cytotoxic lymphocytes generated after repeated stimulation with autologous LCL were restricted by this molecule. One of the cultures was subjected to various stimulation strategies. A relatively low proportion of HLA A2- and HLA B7-restricted cytotoxic T cells could be detected in the autostimulated cultures. Such cells were enriched at the expense of All-restricted ones by stimulating with allogeneic LCLs which lacked HLA All but expressed A2 or B7. Interestingly, stimulation of the lymphocytes with only allogeneic LCL also generated autoreactive CTLs. Thus, by including or using exclusively allogeneic LCL stimulators, the CTL fractions represented by few cells could be enriched.