Human natural killer cytotoxic factor. Studies on its production, specificity, and mechanism of interaction with target cells.
Human natural killer cytotoxic factor. Studies on its production, specificity, and mechanism of interaction with target cells.
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人类自然杀伤细胞毒因子。
DOI：
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发表时间：
1985
期刊：
影响因子：
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通讯作者：
J. Ortaldo
中科院分区：
文献类型：
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作者：
I. Blanca;R. Herberman;J. Ortaldo
人外周血淋巴细胞在无血清条件下体外培养2天产生一种自然杀伤（NK）细胞毒性因子（NKCF），可选择性杀伤NK敏感靶点。在无血清条件下实现了NKCF的最佳释放，而胎牛血清的存在抑制了该因子的产生和活性。NKCF的机制研究表明，该因子可在6小时内被靶细胞吸附，在额外孵育30-48小时后，无需进一步暴露于NKCF，即可使处理的靶细胞达到最大水平的裂解，如111 I释放微量细胞毒性试验所评估。NKCF对靶细胞的吸附及其细胞毒活性被一些磷酸化糖（甘露糖-6PO 4和葡萄糖-6PO 4）抑制，但不被果糖-6PO 4或非磷酸化糖（甘露糖、葡萄糖、半乳糖）抑制。这些结果表明糖-6PO4在NKCF与NK靶细胞的相互作用水平上的作用。这进一步得到以下发现的支持：衣霉素抑制靶细胞糖基化也抑制NKCF吸收至靶细胞和NKCF的直接杀伤。因此，NKCF似乎是一种大颗粒淋巴细胞产生的因子，其由于与靶细胞膜上的糖基化结构相互作用而产生裂解。进行纯化研究，以开始人NKCF的生化表征。结果表明，NKCF的表观分子量在20，000和40，000道尔顿之间。这种方法与放射性标记的NKCF应该是有用的，为进一步研究人类NKCF的生化特性及其作用机制。在无血清条件下引发NKCF的能力应有助于其检测、纯化和生化表征。
Human peripheral blood lymphocytes cultured in vitro for 2 days in serum-free conditions produced a natural killer (NK) cytotoxic factor (NKCF) which selectively killed NK-susceptible targets. Optimal release of NKCF was achieved under serum-free conditions, while the presence of fetal calf serum inhibited both the production and activity of the factor. Mechanistic studies with NKCF demonstrated that the factor could be adsorbed by the target cells within 6 h, with no further exposure to NKCF required for maximal levels of lysis of the treated targets after additional 30-48 h of incubation, as assessed by a 111I release microcytotoxicity assay. NKCF adsorption to target cells and its cytotoxic activity were inhibited by some phosphorylated sugars (mannose-6PO4 and glucose-6PO4), but not by fructose-6PO4 or nonphosphorylated sugars (mannose, glucose, galactose). These results suggest a role of sugar-6PO4 at the level of interaction of NKCF with NK target cells. This was further supported by the finding that inhibition of target cell glycosylation by tunicamycin also inhibited absorption of NKCF to the target cells and direct killing by NKCF. Therefore, it appears that NKCF is a large granular lymphocyte produced factor which produces lysis as a result of the interaction with glycosylated structures on target cell membranes. Purification studies were performed to begin biochemical characterization of human NKCF. The results indicated that NKCF has an apparent molecular weight between 20,000 and 40,000 dalton. Such approaches with radiolabeled NKCF should be useful for the further study of the biochemical characteristics of human NKCF and of its mechanism of action. The ability to elicit NKCF under serum-free conditions should facilitate its testing, purification, and biochemical characterization.