Human peripheral lymphocytes in tissue culture synthesize immunoglobulin Ig which is secreted into the medium. The small amounts of Ig produced can be detected in nanogram quantities by a modification of the ammonium sulfate test. Ig formation by peripheral human lymphocytes was affected by hydrocortisone when the cells were briefly exposed to the hormone at the onset of incubation. When hydrocortisone was dissolved in culture medium or when the more soluble hydrocortisone-21-sodium succinate was employed, significantly increased in vitro synthesis of Ig was noted at 10––9 to 10––5 and 10––10 to 10––6 M, respectively (both p < 0.01). With hydrocortisone-21-sodium succinate, in the concentration range 10––5 to 10––3M, no stimulation of Ig synthesis was observed and at 10––3 M there was decreased Ig synthesis compared to the controls. Average lymphocyte viability at all concentrations was in the range of 39–52% but at the highest concentrations of hydrocortisone tended to be somewhat less, but not significantly. Similar results were obtained when hydrocortisone was dissolved in ethanol but lymphocyte viability was more variable in these experiments. Radioimmunoelectrophoretic and radioimmunodiffusion analysis of culture supernatants from culture including 14C-l-leucine showed that the only Ig formed was IgG; a β-globulin was also formed (radioimmunoelectrophoresis). These experiments demonstrate that human peripheral lymphocytes cultured in vitro interact with hydrocortisone to modify Ig synthesis.