CD28 and cytotoxic T-lymphocyte antigen-4 (CTLA-4), which have opposing functions in T-cell responses, share the two ligands, CD80 (B7-1) and CD86 (B7-2). To understand the functions of CD80 and CD86, respectively, CD80(low) dendritic cells (DCs) and CD86(low) DCs were prepared by using RNA interference. Then CD80 and CD86 functions were analysed by in vitro mixed lymphocyte reaction and cytokine production assay. Effect on cardiac allograft survival was assayed in vivo. In this study, graded stimulatory function of CD80 and CD86, stronger inhibition of proliferation, and stronger prolongation of transplant survival were observed when CD80 and CD86 were blocked simultaneously.