Sadlik J R, Hoyer M, Leyko M A, Horvat R, Parmely M, Whitacre C, Zwilling B, Rinehart J J
Cancer Res. 1985 May;45(5):1940-5.
Recently, gamma-interferon (IFN-gamma) has been shown to be have the capacity to activate macrophages in several murine and human systems. The studies reported here were undertaken to determine the identity of the lymphokine responsible for activation of human monocytes to a tumoricidal state. Macrophage-activating factor (MAF) activity was assessed using a 24-h 51Cr release assay with human monocytes as effector cells and K-562 targets. Stimulated lymphocyte supernatants were produced by stimulation of peripheral blood mononuclear cells with concanavalin A in serum-free media. Interferon was detected in an antiviral assay. Four lines of evidence lead to the conclusion that MAF and IFN-gamma are identical in this system: (a) fractionation of stimulated lymphocyte supernatants by adsorption chromatography, followed by anion or cation exchange chromatography (Mono-S, Mono-Q columns), resulted in nearly identical elution profiles of MAF and IFN activities. All of the individual fractions containing MAF activity were found to contain IFN in amounts corresponding to MAF activity. (b) Monoclonal antibody specific for IFN-gamma neutralized the ability of stimulated lymphocyte supernatants to induce human monocyte tumoricidal activity. This antibody also neutralized the MAF activity of purified IFN-gamma but not alpha-interferon. (c) The biological MAF activity of activated lymphocyte supernatants and IFN-gamma were similar. Dilution versus MAF activity for IFN-gamma and stimulated lymphocyte supernatants exhibited identical slopes. Lymphocyte supernatants and IFN-gamma demonstrated similar MAF activity on three effector cells: monocytes, in vitro-differentiated macrophages, and dexamethasone-differentiated macrophages. (d) Analysis of supernatants produced by five antigen-stimulated human T-cell clones demonstrated coordinate production of MAF and IFN. These results provide compelling evidence for support of the concept that IFN-gamma is the major human lymphokine capable of inducing monocyte-macrophage tumoricidal activity.
最近，γ干扰素（IFN-γ）已被证明在多个小鼠和人类系统中具有激活巨噬细胞的能力。本文所报道的研究旨在确定负责将人类单核细胞激活至杀肿瘤状态的淋巴因子的身份。巨噬细胞激活因子（MAF）活性通过以人类单核细胞作为效应细胞、K-562细胞为靶细胞的24小时51Cr释放试验进行评估。刺激淋巴细胞上清液是通过在无血清培养基中用刀豆球蛋白A刺激外周血单核细胞产生的。通过抗病毒试验检测干扰素。四条证据链得出结论：在该系统中MAF和IFN-γ是相同的：（a）通过吸附色谱法对刺激淋巴细胞上清液进行分级分离，随后进行阴离子或阳离子交换色谱法（Mono-S、Mono-Q柱），MAF和IFN活性的洗脱曲线几乎相同。所有含有MAF活性的单个组分都被发现含有与MAF活性相对应量的IFN。（b）对IFN-γ特异的单克隆抗体中和了刺激淋巴细胞上清液诱导人类单核细胞杀肿瘤活性的能力。该抗体也中和了纯化的IFN-γ的MAF活性，但不中和α干扰素的MAF活性。（c）活化淋巴细胞上清液和IFN-γ的生物学MAF活性相似。IFN-γ和刺激淋巴细胞上清液的稀释度与MAF活性呈现相同的斜率。淋巴细胞上清液和IFN-γ在三种效应细胞上表现出相似的MAF活性：单核细胞、体外分化的巨噬细胞和地塞米松分化的巨噬细胞。（d）对五个抗原刺激的人类T细胞克隆产生的上清液的分析表明MAF和IFN是协同产生的。这些结果为支持IFN-γ是能够诱导单核细胞-巨噬细胞杀肿瘤活性主要人类淋巴因子这一概念提供了有力证据。
