Evaluation of in vivo tolerogenicity of genetically modified recipient dendritic cells (syngeneic DC) pulsed with immunogenic peptides (allopeptides) derived from donor HLA molecule in HLA class I transgenic mouse.

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T cells of C3H.B51 recognize HLA-B^*3501 molecules expressed on C3H.B35 as allo-MHC class I antigens and rejects vascularized C3H. B35 grafts by cellular mechanism. HLA-B^*3501 derived peptide induces long-term heart graft survival in C3H.B35 into C3H.B51 cardiac transplantation model by intra-thymic injection. (Transplant proceedings, 30, 3890-3891, 1998.) We plan to clarify the putative tolerogenic capability of DC involved with HLA molecules based on this HLA TGM heart transplant model. Application of syngeneic (recipient : C3H.B51) DC exposed to allo-antigen (HLA-B*3501 derived peptides) with some modification is expected to establish donor(C3H.B35) specific tolerance. C3H.B35 and C3H.B51 HLA transgenic mouse : So far, after several months of breeding management, the specific TGM strains have been successfully recovered from cryo-preservation. The allogeneic response, which is the key factor in this study, has been confirmed by both in vitro and in vivo study. In brief, mean survival of skin graft from C3H.B35 on C3H.B51 was 15 days and survival of heterotopic heart graft was 24 days. These findings coincides with our previous report (Ando and Beck et al., Transplantation 68, 904-908, 1999.). C3H.B35 specific cytotoxic T cell was generated from splenocytes obtained from C3H.B51 primed by C3H.B35 skin grafting and co-cultured with irradiated C3H.B35 splenocytes. The specific %lysis was more than 50% at the E : T ration of 80 : 1. DC : DC obtained from mouse bone marrow cells after removal of adherent cells followed by GM-CSF and IL4 addition showed marked allo-stimulatory effect in mixed leukocyte reaction.We plan to investigate the possibility of regulating the allo-immune-response between the TGM strain by modified DC propagated from TGM.