The effects of purified mitogenic proteins (Pa-1 and Pa-2) from pokeweed on human T and B lymphocytes in vitro.

Purified proteins (Pa-1 and Pa-2) from pokeweed have been compared with commercial pokeweed mitogen (PWM-G) and other mitogens in their ability to stimulate human lymphocytes. With cultures of T and B cells separated from tonsil lymphocytes, thymidine uptake, blast transformation and immunoglobulin (Ig) synthesis have been measured. IgM and IgG was measured in supernates of stimulated cultures by radioimmunoassay. Pa-1, Pa-2 and PWM-G were found to be potent mitogens for unseparated tonsil lymphocytes or nylon column purified T cells. Pa-2 was found to be active at lower concentrations than Pa-1, and PWM-G was less potent than the purified mitogens. These three mitogens all stimulated unseparated lymphocytes to secrete large quantities of Ig (20-100 mug/ml) during 7 days in culture. With increasing amounts of mitogens severe decreases in immunoglobulin synthesis were observed at day 6 even with doses which were still optimal for stimulation of thymidine uptake at days 3 and 6. With purified B cells (less than 2% T cells) Pa-1 was the best mitogen for thymidine incorporation. However, the secretory response was very variable. In some experiments B cells did not secrete Ig in response to mitogens; in others Pa-1 was clearly more effective at stimulating secretion than Pa-2 or PWM-G and in some experiments B cells were stimulated by all three. In one experiment Pa-1 stimulated prolymphocytic leukaemia cells to blast transformation and the secretion of IgM. It is concluded that Pa-1, Pa-2 and PWM-G are much better activators of Ig synthesis in human cultures than either PHA or LPS and that Pa-1 is the most reliable B-cell stimulant of the three.