Matsuyama H, Yoshihiro S, Ohmoto Y, Yamamoto N, Naito K
Department of Urology, Yamaguchi University School of Medicine, Ube, Japan.
Cancer Immunol Immunother. 1993 Jul;37(2):84-8. doi: 10.1007/BF01517039.
A highly purified natural alpha-interferon (nIFN alpha) was tested in vitro for direct and indirect antiproliferative activity against renal cell carcinoma (RCC), using a modified human tumor clonogenic assay and clinically achievable concentrations. In preclinical experiments, the indirect (cytokine-mediated) antiproliferative activity of nIFN alpha was investigated using ACHN cells (established human RCC cell line). Continuous exposure to nIFN alpha at concentrations of more than 5 IU/ml in the presence of feeder cells (a mixture of 5 x 10(4) monocytes/dish and 5 x 10(5) lymphocytes/dish, obtained from healthy donors) significantly inhibited colony formation of ACHN cells in comparison with growth inhibition in the absence of feeder cells (P < 0.05). Various cytokines were measured in the supernatants lying over the medium on the feeder-layer agarose containing the same conditioned feeder cells. With IFN alpha at 500 IU/ml, tumor necrosis factor alpha (TNF alpha) and IFN gamma were detected at markedly high levels for 2-24 h. Neutralizing anti-TNF alpha monoclonal antibody significantly reduced the indirect antiproliferative activity. Using our modified human tumor clonogenic assay technique, sufficient numbers of colonies for drug testing were observed in 19 of 31 surgical specimens (61.3%). In these clinical materials, nIFN alpha at a clinically achievable concentration (50 IU/ml) significantly inhibited colony growth in the presence of feeder cells consisting of 5 x 10(4) monocytes/dish and 5 x 10(5) lymphocytes/dish, obtained from the patient whose tumor was examined (P < 0.05). In colony-forming cases, a significant correlation between the percentage colony survival and TNF alpha concentration in the supernatant was observed (r = -0.95, P < 0.01). These results suggest that this assay system may be an appropriate technique for evaluating the antiproliferative activities of nIFN alpha involving cytokine-mediated action, and that TNF alpha may play an important role in this cytokine-mediated activity.
使用改良的人肿瘤克隆形成试验和临床可达到的浓度，在体外测试了一种高度纯化的天然α-干扰素（nIFNα）对肾细胞癌（RCC）的直接和间接抗增殖活性。在临床前实验中，使用ACHN细胞（已建立的人肾癌细胞系）研究了nIFNα的间接（细胞因子介导的）抗增殖活性。在存在饲养细胞（从健康供体获得的5×10⁴个单核细胞/培养皿和5×10⁵个淋巴细胞/培养皿的混合物）的情况下，以超过5 IU/ml的浓度持续暴露于nIFNα，与在无饲养细胞时的生长抑制相比，显著抑制了ACHN细胞的集落形成（P<0.05）。在含有相同条件饲养细胞的饲养层琼脂糖上的培养基上层清液中测量了各种细胞因子。当IFNα浓度为500 IU/ml时，在2至24小时内检测到肿瘤坏死因子α（TNFα）和IFNγ的水平明显升高。中和抗TNFα单克隆抗体显著降低了间接抗增殖活性。使用我们改良的人肿瘤克隆形成试验技术，在31个手术标本中的19个（61.3%）中观察到了足够数量用于药物测试的集落。在这些临床材料中，临床可达到的浓度（50 IU/ml）的nIFNα在存在由所检查肿瘤患者获得的5×10⁴个单核细胞/培养皿和5×10⁵个淋巴细胞/培养皿组成的饲养细胞时，显著抑制了集落生长（P<0.05）。在形成集落的病例中，观察到集落存活率百分比与上清液中TNFα浓度之间存在显著相关性（r = -0.95，P<0.01）。这些结果表明，该检测系统可能是评估涉及细胞因子介导作用的nIFNα抗增殖活性的合适技术，并且TNFα可能在这种细胞因子介导的活性中起重要作用。
