Staphylococcal enterotoxins, like several plant lectins, have been shown to stimulate mitogenic activity and lymphokine production in lymphocytes. The effect of enterotoxins A and B on the primary in vitro plaque-forming cell (PFC) response of mouse (C57BL/6) spleen cells to sheep red blood cells (SRBC) was examined. PFC responses in 4- or 5-day cultures were inhibited greater than 90% by 0.1 mug enterotoxin A and by 3- to 6-mug enterotoxin B when the toxins were added to the cultures either at the time of SRBC addition or 24 hr before. Both enterotoxins A and B were shown to be T lymphocyte mitogens. Kinetic data indicated that the enterotoxins (10 mug) inhibited the PFC response as early as day 3, but were relatively more inhibitory at days 4 and 5. The PFC response was inhibited when enterotoxins were added to cultures at the same time as antigen or a day later; it was enhanced when added at day 2; and it was unaffected when added at day 3 of 4-day cultures. Further, the presence of enterotoxin A during the first 24 hr of culture and subsequent removal was still as effective in inhibiting the PFC response as when it was present throughout the culture period. The PFC inhibitory properties of enterotoxins appear, then, to affect some early event(s) in the in vitro immune response. Alpha-methyl-D-mannopyranoside blocked the PFC inhibitory effect of concanavalin A (con A), but it had no effect on enterotoxin A. The two mitogens appear, then, to react with different receptors on the lymphocytes or to differ in the dynamics of their reactivity. The effects of staphylococcal enterotoxins on the PFC response of spleen cells were remarkably similar to those reported for lectins such as con A. Enterotoxins are structurally simpler than con A and should therefore be quite useful in studying various biologic activities of lymphocytes.