We have described techniques for induction of primary and secondary human immune responses in vitro to lymphoid cells modified with trinitrophenyl, dinitrophenyl, and fluorescein isothiocyanate. Optimal secondary proliferative responses required the presentation of hapten on stimulator cells that shared HLA-D region determinants with the responder cell and/or the original stimulator cell. In contrast, hapten-specific cytotoxic responses assessed on modified allogeneic targets with no detected HLA homology with the responder were comparable in magnitude to those detected on modified autologous targets. Furthermore, secondary proliferative, but not cytotoxic, responses required presentation of Ia+ stimulator populations. Modified B cells, surface-immunoglobulin-negative, non T cells (null-cells), and Ia+ activated T cells all induced proliferative responses at least as effectively as equal numbers of hapten-conjugated macrophage/monocytes. Conversely, Ia(-) null cells and macrophages were entirely unable to stimulate. The data thus suggest that for proliferative responses, primed human T cells respond to modified lymphoid cells only when hapten is recognized in the context of Ia molecules.