The immunomodulatory effects of thymosins on the Ag-presenting capacity of macrophages were investigated. Using an in vitro antigen (Ag)-specific macrophage-dependent T-cell proliferation system, we found that both thymosin alpha 1 (T alpha 1) and thymosin beta 4 (T beta 4) augment the Ag-presenting capacity of macrophages. Macrophage monolayers were pulsed with keyhole limpet hemocyanin (KLH) in the absence or presence of thymosins, washed and overlaid with spleen cells. Splenocytes were collected, mitomycin C-treated and injected into syngeneic mice. Draining lymph node cells were tested for Ag-specific response by measuring proliferation, interleukin 2 (IL-2) secretion and expression of IL-2 receptors (IL-2R) on their cell surface. We found that the presence of thymosins during the pulsing of macrophages with KLH led to significantly enhanced lymph node cell proliferation responses to KLH was correlated to increased IL-2 production and IL-2R expression. The concentrations of T alpha 1 and T beta 4 required for amplification were 10(-8) to 10(-10) M, well within the physiological range of activity of most peptide hormones. The observed enhancement of IL-2 secretion was not accompanied by interleukin 4 (IL-4) production. This study is the first to demonstrate that thymic hormones have the ability to increase the efficiency of antigen presentation by macrophages. The results suggest that an initial step in the regulation of the immune function by T alpha 1 and T beta 4 may involve activation of the macrophages at the time of antigen presentation.