Stimulation natural cytotoxic activity in mixed cell cultures and by culture supernatants containing interferon.
Stimulation natural cytotoxic activity in mixed cell cultures and by culture supernatants containing interferon.
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DOI：
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发表时间：
1982
期刊：
影响因子：
6.4
通讯作者：
M. Potter;M. Moore;A. Morris
中科院分区：
文献类型：
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作者：
M. Potter;M. Moore;A. Morris
我们之前已经证明，用多种不同的丝裂霉素-C 处理的淋巴细胞系培养 5 天的淋巴细胞制剂显示出非特异性（NK 样）细胞毒性活性的增加。我们现在检查了通过绵羊红细胞 (SRBC) 玫瑰花结分离制备的富含 T 细胞和 B 细胞的应答群体中诱导的活性，并发现增加的细胞毒活性主要存在于富含 T 细胞的部分中。为了研究可溶性因子是否参与介导增强的细胞毒性，在测量针对 K562 细胞的细胞毒性之前，使用混合细胞培养物的上清液预处理新鲜分离的效应细胞群。用混合细胞培养物培养 2-6 天的上清液预处理的效应细胞显示出增加的细胞毒性活性，用来自 4 和 5 天共培养物的上清液获得的最大活性。当用混合细胞培养物的上清液预处理效应细胞 2、4 和 18 小时时，观察到活性增加，无论刺激细胞的性质如何（T 细胞和 B 细胞衍生系、K562 和正常同种异体淋巴细胞）。活性上清液是由富含 T 细胞的应答群体产生的，而不是富含 B 细胞的应答群体产生的。还检查了混合细胞培养物的上清液中是否存在干扰素 (IFN)，一般来说，那些介导细胞毒活性大幅增加的上清液也含有显着的抗病毒活性。检测到的IFN不被IFN-β抗血清中和，但被IFN-α抗血清部分中和。 pH 2 处理也消除了部分活性，而 pH 2 和抗-α 的组合完全中和了活性，表明存在 IFN-γ 和 IFN-α。讨论了干扰素作为该系统中增强细胞毒性介质的候选资格。
We have previously demonstrated that lymphocyte preparations cultured for 5 days with a number of different mitomycin-C-treated lymphoid cell lines showed increased non-specific (NK-like) cytotoxic activity. We have now examined the activity induced in T- and B-cell-enriched responder populations, prepared by sheep red blood cell (SRBC) rosette separation, and found that increased cytotoxic activity was present mainly in the T-cell-enriched fraction. To investigate whether soluble factors were involved in the mediation of enhanced cytotoxicity, supernatants from mixed cell cultures were used to pretreat freshly isolated effector cell populations before measurement of cytotoxicity against K562 cells. Effector cells pretreated with supernatants from mixed cell cultures incubated for 2–6 days showed increased cytotoxic activity, the greatest activity being obtained with supernatants from 4 and 5 day co-cultures. Increased activity was observed when effector cells were pretreated for 2, 4 and 18 hr with supernatants from mixed cell cultures, regardless of the nature of the stimulator cell (T- and B-cell-derived lines, K562 and normal allogeneic lymphocytes). Active supernatants were produced by T-cell-enriched, but not B-cell-enriched responder populations. Supernatants from mixed cell cultures were also examined for the presence of interferon (IFN) and in general, those mediating large increases in cytotoxic activity also contained significant anti-viral activity. The IFN detected was not neutralized by antiserum to IFN-β but was partly neutralized by antiserum to IFN-α. pH 2 treatment also removed part of the activity while a combination of pH 2 and anti-α completely neutralized the activity suggesting the presence of IFN-γ and IFN-α. The candidacy of IFNs as mediators of enhanced cytotoxicity in this system is discussed.