This study investigated the effects of carotenoid and capsaicin constituents of Capsicum on intestinal immune responses in mice. Peyer's patch (PP) cells were isolated from mice orally administered with capsaicin, or one of three carotenoids (beta-carotene, beta-cryptoxanthin, or lycopene), at 5 mg/kg/day for 7 consecutive days. Collagenase-separated PP cells were then cultured in the presence or absence of concanavalin A (Con A). PP cells from mice treated with capsaicin, beta-carotene, or beta-cryptoxanthin all showed significantly enhanced interleukin (IL)-2 and interferon (IFN)-gamma production when costimulated with 5 microg/mL Con A, with capsaicin having the greatest effect (approximately two times greater than in normal mice). No increase in the production of IL-2 or IL-4 was observed when PP cells from mice were cultured without Con A. We further tested the combined efficacy of carotenoids and capsaicin on intestinal T-cell cytokine production. Oral administration of capsaicin with beta-carotene, both at 5 mg/kg/day for 7 days, increased IFN-gamma and IL-2 production in cultured PP cells costimulated with Con A. In contrast, oral administration of beta-cryptoxanthin counteracted the stimulatory effect of capsaicin treatment on T-helper cytokine production. Flow cytometric analysis revealed that the population of IFN-gamma(+) and IL-4(+) cells in PPs from mice administered capsaicin and/or carotenoids did not change, which suggested that the effects of carotenoids and capsaicin on cytokine production were not due to changes in the lymphoid population in PPs. These results indicate that carotenoids and capsaicin, which are common components of foods such as Capsicum, mutually modulate T-cell immune responses to exogenous or endogenous inducers such as antigens in PPs, without changing the lymphoid population. Carotenoids modulate the potentiality of cytokine production in T cells or indirectly activate T cells but have no triggering effect such as Con A.