Cytokines produced by blood mononuclear cells stimulated with the streptococcal preparation OK-432: effect on production by supplementing the medium with xenogeneic serum

Abstract

Human blood MNC were stimulated for 7 days with a streptococcal preparation, OK-432, in a medium supplemented with either 10% autologous serum (autoserum) or 10% FCS. Cytokines, including IL-2, IL-1, IFN, and TNF in the supernatants, and antitumor cytotoxicity of MNC were analyzed. None of the IL-2 was detectable during the culture in medium with autoserum, although significant enhancement of cytotoxicity was observed at day 1. On the other hand, production of IL-2 and higher cytotoxicity were induced in the medium with FCS. Even a control culture of MNC unstimulated with OK-432 in the medium with FCS, showed a slight but significant amount of IL-2 and considerable cytotoxicity. However, such a culture in the medium with autoserum showed no such IL-2 production or cytotoxicity. The cytotoxicity induced in the medium with FCS was significantly inhibited by the addition of anti-IL-2 antibody to the culture as well as by the addition of anti-IFN gamma antibody, whereas the cytotoxicity in the medium with autoserum was not inhibited at all by anti-IL-2 antibody. Therefore, the cytotoxicity induced in the medium with FCS included IL-2-induced killer activity, i.e., LAK activity. Similarly, the levels of IL-1, IFN, and TNF production in the supernatants were variable depending on the serum used to supplement the medium. The nonspecific production of IL-2 and the unexpected induction of cytotoxicity were consistently provoked in the medium using several different lots of FCS, but not in the medium with human allogeneic sera or pooled AB serum nor in that with autoserum. It was revealed that the source of sera used to supplement the medium is an important factor affecting the results of analysis of cytokine production and cytotoxicity of MNC induced by certain stimulants.