1, 25-Dihydroxyvitamin D3 [1, 25-(OH)2D3], like the immune response modulators prostaglandin E2 (PGE2) and histamine, inhibits mitogen-induced proliferation of human peripheral blood mononuclear cells. 1, 25-(OH)2D3 acts synergistically with PGE2 and histamine to inhibit lymphocyte mitogenesis. This is apparent at a wide concentration range of 1, 25-(OH)2D3 (3 × 10−11−10−8 mol/L). Cholera toxin, forskolin, and isobutylmethylxanthine, which like PGE2 and histamine increase intracellular concentrations of cAMP, also act synergistically with 1, 25-(OH)2D3 in this system. Culture of mitogenstimulated adherent cell-depleted mononuclear cells with PGE2 increases the number of high affinity binding sites for 1, 25-(OH)2D3. This finding may account for the synergistic interaction between the two agents. © 1990 by The Endocrine Society.