The effects of beta-carotene, retinol, and retinoic acid on function of mononuclear cells during the peripartum period was assessed in vitro. Blood was collected from 14 Holstein cows on wk -4, -1, 0, 1, and 4 postpartum, and mononuclear cells were obtained by gradient centrifugation. Mononuclear cell proliferation induced by concanavalin A was measured in the presence of beta-carotene, retinol (1 x 10(-9) and 1 x 10(-8) M), and retinoic acid (1 x 10(-10) and 1 x 10(-9) M). Retinol and beta-carotene had no effect on spontaneous cell proliferation, whereas retinoic acid was suppressive. However, 1 x 10(-9) M beta-carotene enhanced concanavalin A-induced proliferation at wk -1, whereas 1 x 10(-8) M beta-carotene was suppressive at wk -4. Retinoic acid suppressed concanavalin A-induced proliferation at wk 0, but retinol had no effect. These results suggest a mechanism by which beta-carotene affords the mammary gland protection against infection immediately prepartum.