Hall R E, Muchmore A V, Decker J M, Blaese R M
Cell Immunol. 1983 Feb 15;76(1):58-68. doi: 10.1016/0008-8749(83)90348-9.
In an attempt to develop a constant and reproducible in vitro system for a detailed analysis of cytotoxic effector mechanisms of nonimmune mononuclear phagocytes, the HL-60 promyelocytic cell line was studied for its cytotoxic action on chicken erythrocyte target cells. HL-60 cells cultured in complete medium were found to be noncytotoxic for chicken erythrocytes in an 18-hr 51Cr-release assay. These cells have been shown to acquire several characteristics of mature macrophages upon incubation with phorbol myristate acetate (PMA), and when PMA was included in the medium during the assay, the HL-60 cells became strongly cytotoxic to the target cells in the absence of exogenous antibody, lectin, or serum complement. Freshly isolated peripheral blood monocytes also became cytotoxic in the presence of PMA, whereas peripheral blood lymphocytes and the U937 histiocytic cell line did not. Detectable target lysis was observed between 4 and 8 hr after HL-60 stimulation with PMA, and HL-60 cells prestimulated with PMA for 24 hr retained their cytotoxic activity following washing and assay in PMA-free medium. Cytotoxic HL-60 cells developed after exposure to 10(-6) to 10(-9) M PMA, and significant target cell lysis occurred at effector:target cell ratios as low as 0.5:1. The PMA-induced HL-60-mediated cytotoxic response was markedly inhibited by blockers of protein synthesis, inhibition of microfilament function, and depletion of cellular superoxide and hydrogen peroxide. Interestingly, cytotoxicity of HL-60 cells for chicken erythrocyte targets was modulated by the direct addition of certain simple saccharides to the assay in a fashion similar to that observed with spontaneously cytotoxic mononuclear cells from several vertebrate and invertebrate species. Thus, the cytolytic effector function induced in HL-60 cells by incubation with PMA presents a useful model for the study of cellular cytotoxic mechanisms as well as the mechanisms utilized by nonimmune cells in the recognition of non-self.
为了开发一种稳定且可重复的体外系统，用于详细分析非免疫单核吞噬细胞的细胞毒性效应机制，研究了HL-60早幼粒细胞系对鸡红细胞靶细胞的细胞毒性作用。在18小时的51Cr释放试验中，发现培养在完全培养基中的HL-60细胞对鸡红细胞无细胞毒性。这些细胞在与佛波酯（PMA）孵育后已显示出获得成熟巨噬细胞的若干特征，并且当在试验期间培养基中加入PMA时，HL-60细胞在没有外源性抗体、凝集素或血清补体的情况下对靶细胞具有强烈的细胞毒性。新鲜分离的外周血单核细胞在PMA存在下也变得具有细胞毒性，而外周血淋巴细胞和U937组织细胞系则没有。在用PMA刺激HL-60后4至8小时观察到可检测到的靶细胞裂解，并且用PMA预刺激24小时的HL-60细胞在洗涤并在无PMA的培养基中进行试验后仍保留其细胞毒性活性。暴露于10(-6)至10(-9) M PMA后产生细胞毒性HL-60细胞，并且在效应细胞：靶细胞比率低至0.5:1时发生显著的靶细胞裂解。蛋白质合成阻滞剂、微丝功能的抑制以及细胞超氧化物和过氧化氢的消耗显著抑制了PMA诱导的HL-60介导的细胞毒性反应。有趣的是，通过向试验中直接添加某些单糖，HL-60细胞对鸡红细胞靶标的细胞毒性以类似于在几种脊椎动物和无脊椎动物物种的自发细胞毒性单核细胞中观察到的方式受到调节。因此，通过与PMA孵育在HL-60细胞中诱导的溶细胞效应功能为研究细胞毒性机制以及非免疫细胞在识别非自身中利用的机制提供了一个有用的模型。
