Rapid killing of actinomycin D-treated tumor cells by human mononuclear cells. I. Effectors belong to the monocyte-macrophage lineage.
Rapid killing of actinomycin D-treated tumor cells by human mononuclear cells. I. Effectors belong to the monocyte-macrophage lineage.
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人单核细胞快速杀死放线菌素 D 处理的肿瘤细胞。
DOI：
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发表时间：
1984
影响因子：
4.4
通讯作者：
Alberto Mantovani
中科院分区：
文献类型：
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作者：
F. Colotta;Giuseppe Peri;Antonello Villa;Alberto Mantovani
将肿瘤细胞与放线菌素D(ActD)预先孵育，在6小时的51Cr释放试验中，使各种小鼠和人的细胞系对未经刺激的人外周血单个核细胞(PBM)具有杀伤作用。选择小鼠WEHI 164肉瘤进行药物依赖细胞毒性(DDCC)分析，是因为该肿瘤具有高水平的杀伤力，并且对自然杀伤(NK)细胞活性具有相当大的耐药性。诱导裂解敏感性的最佳条件为1微克/毫升的ActD预孵育3小时，对经ActD处理的WEHI 164细胞具有细胞毒性的效应细胞是可塑性贴壁的(大于85%的单核细胞)。不附着于塑料和尼龙毛(小于或等于1%单核细胞)的细胞没有明显的DDCC活性。相反，NK细胞对K562细胞的杀伤活性是由非贴壁细胞介导的。当PBM在Percoll设计的一步不连续梯度上浓缩单核细胞(大于90%纯度)时，在单核细胞组分中发现了DDCC活性，并且浓缩的淋巴样细胞组分对经Act D处理的WEHI 164细胞没有细胞毒性。而淋巴细胞对K562细胞的杀伤活性恢复，单核细胞无NK细胞毒作用。在设计用于浓缩大颗粒淋巴细胞(LGL)的六级Percoll梯度分级后，密度较高的淋巴细胞(组分4-6)和密度较低的具有NK活性的LGL(组分2-3)对经Act D处理的WEHI 164肉瘤细胞没有细胞毒性。组分1的DDCC活性下降与单核细胞有关。利用荧光激活的细胞分选仪，根据单抗定义的表面标志对PBM进行分级。DDCC效应细胞呈单核细胞标志(Mo2、UCHM1)阳性，NK细胞(B73.1、HNK1)、T细胞(T11)、B细胞(Leu-10)阴性。外周血单核细胞体外培养5~10天获得的巨噬细胞具有DDCC活性。同样，腹膜和支气管肺泡巨噬细胞对经ActD处理的靶细胞有相当大的细胞毒性，而在这些解剖部位发现很少或没有NK活性。人或小鼠来源的细胞与ActD预先孵育3小时，在6小时的51Cr释放试验中，它们对单核细胞杀伤的敏感性是不同的。在小鼠WEHI 164肉瘤和3T3成纤维细胞系以及人CEM白血病细胞系中观察到高水平的细胞毒作用。单核细胞对ALab乳腺癌(人)和8387肉瘤(人)有微弱(但显著)的细胞毒作用。
Preincubation of tumor cells with actinomycin D (Act D) rendered various murine and human lines susceptible to killing by unstimulated human peripheral blood mononuclear cells (PBM) in a 6-hr 51Cr-release assay. The murine WEHI 164 sarcoma was selected for analysis of drug-dependent cellular cytotoxicity (DDCC) because high levels of killing were detected with this tumor, and it was considerably resistant to natural killer (NK) cell activity. Optimal conditions for induction of susceptibility to lysis included a 3-hr preincubation with 1 microgram/ml Act D. Effector cells of cytotoxicity against Act D-treated WEHI 164 cells were plastic adherent (greater than 85% monocytes). Cells nonadherent to plastic and nylon wool (less than or equal to 1% monocytes) had no appreciable DDCC activity. In contrast NK activity against K562 cells was mediated by nonadherent cells. When PBM were fractionated on a one step discontinuous gradient of Percoll designed to enrich for monocytes (greater than 90% pure), DDCC activity was found in the monocyte fraction, and the lymphoid cell-enriched fraction had no cytotoxicity against Act D-treated WEHI 164 cells. In contrast, NK activity against K562 was recovered with lymphoid cells, and monocytes had no NK cytotoxicity. Upon fractionation on a six step Percoll gradient designed to enrich for large granular lymphocytes (LGL), the denser lymphocytes (fraction 4-6) and the less dense LGL with NK activity (fraction 2-3) had no cytotoxicity against Act D-treated WEHI 164 sarcoma cells. DDCC activity sedimented in fraction 1 in association with monocytes. PBM were fractionated according to monoclonal antibody-defined surface markers by using a fluorescence-activated cell sorter. Effector cells of DDCC were positive for monocyte markers (Mo2, UCHM1) and were negative for NK cell (B73.1, HNK1), T cell (T11), and B cell (Leu-10) markers. Macrophages obtained by culturing blood monocytes in vitro for 5 to 10 days had DDCC activity. Similarly, peritoneal and bronchoalveolar macrophages had considerable cytotoxicity against Act D-treated target cells, whereas minimal or no NK activity was found at these anatomic sites. Cells of human or murine origin, preincubated with Act D for 3 hr, were heterogeneous in their susceptibility to monocyte killing in a 6-hr 51Cr-release assay. High levels of cytotoxicity were observed with the murine WEHI 164 sarcoma and 3T3 "fibroblast" line and with the human CEM leukemia. Monocytes were weakly (but significantly) cytotoxic against the ALAB breast carcinoma (human) and the 8387 sarcoma (human).(ABSTRACT TRUNCATED AT 400 WORDS)