Summary
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-IFNγ 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.
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References
Abo T, Balch CM (1981) A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1). J Immunol 127:1024
Abo T, Sugawara S, Amenomori A, Itoh H, Rikiishi H, Moro I, Kumagai K (1986) Selective phagocytosis of Gram-positive bacteria and interleukin 1-like factor production by a subpopulation of large granular lymphocytes. J Immunol 136:3189
Bonavida B, Katz J, Hoshino T (1986) Mechanism of NK activation by OK-432 (streptococcus pyogenes). I. Spontaneous release of NKCF and augmentation of NKCF production following stimulation with NK target cells. Cell Immunol 102:126
Christmas SE, Meager A, Moore M (1986) Studies of the enhancement of natural cytotoxicity by the streptococcal immunopotentiator OK-432. Int J Immunopharmacol 8:83
Golub SH, Golightly MG, Zielske JV (1979) “NK-like” cytotoxicity of human lymphocytes cultured in media containing fetal bovine serum. Int J Cancer 24:273
Ichimura O, Suzuki S, Saito M, Sugawara Y, Ishida N (1985) Augmentation of interleukin 1 and interleukin 2 production by OK-432. Int J Immunopharmacol 7: 263
Ishida N, Saito M, Nanjo M (1984) Induction of LAK cells in peritoneal cavity of mice after IP injection of OK-432 and antitumor effect of these LAK cells. Jpn J Cancer Chemother 11:2681
Katano M, Torisu M (1983) New approach to management of malignant ascites with a streptococcal preparation, OK-432. II. Intraperitoneal inflammatory cell-mediated tumor cell destruction. Surgery 93:365
Kim YB (1985) Effect of biological response modifier-streptococcal preparation OK-432 on NK/K cell system of gnotobiotic miniature swine. Prog Clin Biol Res 181:337
Mihich E (1986) Future perspectives for biological response modifiers: a viewpoint. Semin Oncol 13:234
Nara K, Odagiri H, Fujii M, Yamanaka Y, Yokoyama M, Morita T, Sasaki M, Kon M, Abo T (1987) Increased production of tumor necrosis factor and prostaglandin E2 by monocytes in cancer patients and its unique modulation by their plasma. Cancer Immunol Immunother 25:126
Oshima K, Kano S, Takaku F, Okumura K (1980) Augmentation of mouse natural killer cell activity by a streptococcal preparation, OK-432. JNCI 65:1265
Saito M, Ichimura O, Kataoka M, Moriya Y, Ueno K, Sugawara Y, Nanjo M, Ishida N (1986) Pronounced antitumor effect of LAK-like cells induced in the peritoneal cavity of mice after intraperitoneal injection of OK-432, a killed streptococcal preparation. Cancer Immunol Immunother 22:161
Suzuki R, Suzuki S, Igarashi M, Kumagai K (1986) Induction of interleukin 3 but not interleukin 2 or interferon production in the syngeneic mixed lymphocyte reaction. J Immunol 137:1564
Suzuki R, Suzuki S, Takahashi T, Kumagai K (1986) Production of a cytokine with interleukin 3-like properties and cytokine-dependent proliferation in human autologous mixed lymphocyte reaction. J Exp Med 164:1682
Uchida A, Hoshino T (1980) Clinical studies on cell-mediated immunity in patients with malignant disease. I. Effect of immunotherapy with OK-432 on lymphocyte subpopulation and phytomitogen responsiveness in vitro. Cancer 45:476
Uchida A, Klein E (1985) Activation of human blood lymphocytes and monocytes by the streptococcal preparation OK-432: enhanced generation of soluble cytotoxic factors. Immunol Lett 10:177
Uchida A, Michsche M (1981) In vitro augmentation of natural killing activity by OK-432. Int J Immunopharmacol 3:365
Wakasugi H, Kasahara T, Minato N, Hamuro J, Miyata M, Morioka Y (1982) In vitro potentiation of human natural killer cell activity by a streptococcal preparation, OK-432: interferon and interleukin-2 participation in the stimulation with OK-432. JNCI 69:807
Yamamoto A, Nagamuta M, Usami H, Sugawara Y, Watanabe N, Niitsu Y, Urushizaki I (1986) Release of tumor necrosis factor (TNF) into mouse peritoneal fluids by OK-432, a streptococcal preparation. Immunopharmacology 11:79
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Abbreviations used: NK, natural killer; IFN, interferon; TNF, tumor necrosis factor; IL-1, interleukin-1; IL-2, interleukin-2; LAK, lymphokine-activated killer; MNC, mononuclear cells; FCS, fetal calf serum; 3H-TdR, 3H-Thymidine
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Fujii, M., Abo, T. & Kumagai, K. Cytokines produced by blood mononuclear cells stimulated with the streptococcal preparation OK-432: effect on production by supplementing the medium with xenogeneic serum. Cancer Immunol Immunother 27, 97–102 (1988). https://doi.org/10.1007/BF00200011
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DOI: https://doi.org/10.1007/BF00200011