We have shown earlier increased intracellular levels of cAMP in peripheral lymphocytes from HIV-seropositive subjects and that a chemically induced decrease in this level increases cell proliferation and cytotoxicity. Others have shown that serotonin indirectly decreases intracellular cAMP levels in normal peripheral lymphocytes. In this study, we show that addition of serotonin decreases intracellular levels of cAMP in lymphocytes from HIV-seropositive subjects and significantly increases the proliferative capacity in vitro. However, the effect of serotonin varies with the initial proliferative response; e.g., these with the highest initial responses have the highest increases. An increase in IL-2 production may be a part of this mechanism since addition of serotonin to in vitro cultures of PHA-stimulated cells increases the expression of mRNA for IL-2 and IFN-gamma. The effect on lymphocyte proliferation was most likely mediated through the serotonin 5HT1a receptor because similar results could be obtained by using DPAT, a specific activator of this receptor. Changes in the expression of 5HT1a receptors as judged by the expression of mRNA could not explain why serotonin in vitro had a stronger enhancing effect on cell proliferation in some HIV-seropositive individuals than in others.