Mantovani A, Dean J H, Jerrells T R, Herberman R B
Istituto di Ricerche Farmacologiche Mario Negri, Via Eritrea 62, Milan, Italy.
Int J Cancer. 1980 Jun 15;25(6):691-9. doi: 10.1002/ijc.2910250602.
Monocytes were separated from peripheral blood mononuclear cells of normal human donors by adherence on plastic conditioned by cell lines (microexudate-coated plastic) and harvested by exposure to ethylene diamine tetra-acetic acid. Cytolytic activity was tested by incubating effector cells for 48 h with the murine SV40-transformed TU5 kidney line or the human lung cancer-derived CaLu line prelabelled with tritiated thymidine. Lymphokine-containing supernatants were obtained from in vitrocultures of lymphoid cells with phytohemagglutinin (PHA), purified protein derivative (PPD), or with Corynebacterium parvum strains CN6134 or CNS888. The monocytes had significant levels of spontaneous cytotoxicity and exposure to lymphokine supernatants markedly enhanced their tumoricidal activity. Augmentation of monocyte-mediated cytotoxicity required a minimal exposure to lymphokine supernatants for 4 h and was maximal after 24 h of preincubation. Treatment of the effector cells with anti-human T-cell serum and complement did not affect either their spontaneous or their lymphokine-stimulated cytotoxicity, whereas silica impaired both reactivities. Supernatants of cultures with PHA, PPD and C. parvum CN6134 had significant levels of interferon (IF). Since partially purified human fibroblast or leukocyte IF was able to stimulate monocyte-mediated cytotoxicity, the IF in these supernatants could play some role in the stimulation of the monocytes. However, C. parvum CN5888 supernatants, which had little IF, enhanced monocyte cytotoxicity as effectively as the C. parvum CN6134 supernatants, strongly suggesting that lymphocyte mediators other than IF can augment the tumoricidal activity of these effector cells. Mature macrophages obtained by in vitro cultivation of monocytes for 4-7 days retained natural cytolytic activity and showed enhanced cytotoxicity in the presence of lymphokines. However, more prolonged in vitro cultivation (> 10 days) resulted in cultures of epithelioid and multinucleated cells which had little natural cytotoxicity and were not responsive to lymphokines.
通过贴壁于经细胞系处理的塑料（微渗出物包被的塑料）从正常人类供体的外周血单核细胞中分离出单核细胞，并通过暴露于乙二胺四乙酸进行收获。通过将效应细胞与预先用氚化胸腺嘧啶标记的鼠SV40转化的TU5肾细胞系或人肺癌衍生的CaLu细胞系孵育48小时来测试细胞溶解活性。含淋巴细胞因子的上清液从用植物血凝素（PHA）、纯化蛋白衍生物（PPD）或用细小棒状杆菌菌株CN6134或CNS888进行体外培养的淋巴细胞培养物中获得。单核细胞具有显著水平的自发细胞毒性，并且暴露于淋巴细胞因子上清液显著增强了它们的杀肿瘤活性。单核细胞介导的细胞毒性增强需要至少暴露于淋巴细胞因子上清液4小时，并且在预孵育24小时后达到最大值。用抗人T细胞血清和补体处理效应细胞既不影响它们的自发细胞毒性也不影响它们的淋巴细胞因子刺激的细胞毒性，而二氧化硅损害了这两种反应性。用PHA、PPD和细小棒状杆菌CN6134培养的上清液具有显著水平的干扰素（IF）。由于部分纯化的人成纤维细胞或白细胞IF能够刺激单核细胞介导的细胞毒性，这些上清液中的IF可能在刺激单核细胞中发挥某种作用。然而，几乎没有IF的细小棒状杆菌CN5888上清液与细小棒状杆菌CN6134上清液一样有效地增强了单核细胞的细胞毒性，强烈表明除IF之外的淋巴细胞介质可以增强这些效应细胞的杀肿瘤活性。通过体外培养单核细胞4至7天获得的成熟巨噬细胞保留了天然细胞溶解活性，并在存在淋巴细胞因子的情况下显示出增强的细胞毒性。然而，更长时间的体外培养（> 10天）导致上皮样和多核细胞培养物，其具有很少的天然细胞毒性并且对淋巴细胞因子无反应。
