Inactivated, partially purified simian immunodefiency virus (SIV(mac)) protected macaques from intravenous challenge with homologous and heterologous strains of SIV that had been grown on human cells but no protection against challenge with monkey peripheral blood mononuclear cell-grown SIV(mac) was afforded. Human immunodeficiency virus type 1 prepared in an analogous way to the SIV(mac) vaccine on the C8166 human T cell line protected macaques against challenge with human cell-grown SIV(mac). These results suggest that protection may be mediated by xenoimmunization with the vaccine cell substrate proteins. All vaccinated macaques had anti-cell antibodies. Major reactivity to MHC class I antigens was found as well as to a 70-kD protein detectable only under nonreducing conditions.