Genetic analysis of the determinants causing an early proliferation in MLC after priming with allogeneic cells in vivo was performed. The active determinants were localized in the I-A subregion of the H-2 complex by using appropriate recombinant mouse strains. Upon priming with antigens coded by the whole H-2 complex, the lymphocytes gave a strong MLC response only to the priming stimulator cells and cells sharing the I-A subregion determinants of the latter. The response to third party stimulator cells and cells not sharing I-A subregion determinants was low. Upon priming with various H-2 region products, only lymphocytes primed against I-A subregion products gave an early proliferative MLC response to the priming stimulator cells, whereas lymphocytes primed against other H-2 region products gave none.