Ernst M, Lange A, Flad H D, Havel A, Ennen J, Ulmer A J
Eur J Immunol. 1984 Jul;14(7):634-9. doi: 10.1002/eji.1830140710.
Monocyte/macrophage-mediated cytotoxicity requires the generation of activated oxygen radicals, which can be measured by chemiluminescence (CL). To investigate whether natural killer (NK) cell activity required activated oxygen species, both cytotoxicity against K562 target cells and CL were measured in cell populations of human peripheral blood. The following results were obtained: (a) Peripheral blood mononuclear cells (MNC) showed NK activity and a response in CL, which could be induced by viable or paraformaldehyde-fixed K562 target cells as well as by latex particles. (b) Both T cells and non-T cells exhibited NK activity, but T cells gave no K562- or latex-induced CL responses. (c) Depletion of phagocytic cells from MNC abolished CL, but only marginally affected NK activity. (d) Reconstitution of phagocyte-depleted MNC with adherent cells revealed a superadditive enhanced CL response, but had no augmenting effect on NK activity. (e) Phagocyte-depleted cell populations, enriched for NK activity by density gradient centrifugation, did not respond in K562- and latex-induced CL. (f) MNC, highly enriched for NK activity by cell sorting with a cytofluorograf using the fluorescein isothiocyanate-labeled monoclonal antibody anti-Leu-11a, responded only with reduced CL, whereas the NK activity was enriched up to 45-fold. From these results it is concluded that NK cell-mediated cytolysis of K562 target cells and K562-induced CL are not functionally correlated, but represent properties of two distinct cell populations, namely NK cells and monocytes.
单核细胞/巨噬细胞介导的细胞毒性作用需要产生活性氧自由基,而活性氧自由基可通过化学发光(CL)来测定。为了研究自然杀伤(NK)细胞活性是否需要活性氧,我们对人外周血中的细胞群体同时测定了其对K562靶细胞的细胞毒性和化学发光。获得了以下结果:(a)外周血单个核细胞(MNC)表现出NK活性和化学发光反应,活的或经多聚甲醛固定的K562靶细胞以及乳胶颗粒均可诱导该反应。(b)T细胞和非T细胞均表现出NK活性,但T细胞对K562或乳胶诱导的化学发光无反应。(c)从MNC中去除吞噬细胞可消除化学发光,但仅轻微影响NK活性。(d)用贴壁细胞重建去除吞噬细胞的MNC显示化学发光反应超相加增强,但对NK活性无增强作用。(e)通过密度梯度离心富集NK活性的去除吞噬细胞的细胞群体,对K562和乳胶诱导的化学发光无反应。(f)使用异硫氰酸荧光素标记的抗Leu-11a单克隆抗体通过细胞荧光分析仪进行细胞分选,高度富集NK活性的MNC仅表现出降低的化学发光反应,而NK活性则富集了45倍。从这些结果可以得出结论,NK细胞介导的对K562靶细胞的细胞溶解作用与K562诱导的化学发光在功能上不相关,而是代表两个不同细胞群体即NK细胞和单核细胞的特性。