Further studies on the mechanism of suppression of human lymphocyte transformation by human alpha fetoprotein

Abstract

We investigated the possibility that noncovalent binding of negatively charged molecules such as prostaglandins (PGs) might be partly responsible for human alpha fetoprotein (HAFP) heterogeneity with respect to charge and the suppression of human lymphocyte responses, since PGs are potent suppressors of lymphocyte transformation. Indomethacin, an inhibitor of PG synthesis, had no effect upon the transformation of adherent-cell-depleted human lymphocytes, nor did it interfere with the capacity of HAFP to inhibit lymphocyte transformation. When a partially suppressive dose of HAFP was added to mitogen-stimulated lymphocytes together with varying doses of prostaglandin E 1 or E 2 (PGE 1 or PGE 2 ), no evidence of inhibitory synergy was demonstrable; their combined suppressive action was either less than or approximately equal to the sum of their respective inhibitory effects. Addition of PGE 2 to a nonsuppressive dose of an impotent HAFP preparation resulted in no greater lymphocyte suppression than that achieved by PGE 2 alone. Analysis of the kinetics of suppression of lymphocyte transformation by PG and HAFP yielded unequivocal evidence for their disparate mechanisms of action. Suppression of lymphocyte transformation by PGE 2 is evident by 24 to 40 hr of culture, persists throughout the entire culture period (including peak deoxyribonucleic acid [DNA] synthesis at 72 to 88 hr and beyond), and does not vary in profundity at any time. By contrast, HAFP-induced suppression of lymphocyte DNA synthesis is lacking at 24 to 40 hr, reaches a peak at 72 to 88 hr, and is waning by 96 to 112 hr. Cultures containing HAFP synthesize more DNA than control cultures during the 120 to 136 hr culture period. These observations, together with our earlier demonstration of a subpopulation of human lymphocytes resistant to HAFP inhibition, suggest that, after the first 24 to 40 hr of culture, HAFP may enhance the proliferation of a suppressor population which gradually dampens the proliferative response. HAFP is capable of suppressing the proliferative response of periodate-treated lymphocytes; this confirms that it does not act via competition with the cell membrane for a mitogen-binding site. Immunofluorescence studies indicate that human peripheral blood lymphocytes lack membrane-associated HAFP or HAFP receptors of high affinity. Our results indicate that PG, either present as a putative contaminant in HAFP isolates or endogenously synthesized by lymphocytes (or monocytes), plays no role in lymphocyte suppression by HAFP.