The oral administration of soluble protein antigen results in profound immunological tolerance. However, the tissue location and function of antigen-presenting cells (APC) that stimulate this response remain unclear. We have hypothesized that the properties of cells presenting antigen to naive T cells within the gut are involved, and therefore gut APC should stimulate T-cell responses with different characteristics to those induced by other APC. To test this, we studied in vitro primary T-cell responses following presentation of soluble protein antigen by cells from the Peyer's patches (PPC) and lamina propria (LPC) of the murine small intestine and the spleen (SPLC). Each APC population stimulated antigen-specific proliferative responses with similar anamnestic characteristics; however, analysis of the cytokines produced revealed marked differences. Whereas SPLC stimulated the balanced production of T-helper type 1 (Th1) and Th2 cytokines, PPC induced a profile consistent with the provision of T-cell help for IgA production. Interestingly, presentation of antigen by LPC stimulated high levels of interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) in the absence of other cytokines [interleukin-2 (IL-2), IL-4, IL-5]. Evidence from analysis of cell activation and division within the cultures suggested that this profile may result from the preferential activation of CD8+ T cells by LPC; however, the lack of conventional CD4+ T-cell cytokines indicated a defect in the normal function of these cells. Adoptive transfer of antigen-pulsed LPC to syngeneic animals abrogated the induction of delayed-type hypersensitivity (DTH) responsiveness, which followed a subsequent conventional antigen challenge further suggesting a role for lamina propria APC in tolerance induction.