Gastl G A, Abrams J S, Nanus D M, Oosterkamp R, Silver J, Liu F, Chen M, Albino A P, Bander N H
Department of Surgery/Urology, New York Hospital-Cornell Medical Center, NY 10021.
Int J Cancer. 1993 Aug 19;55(1):96-101. doi: 10.1002/ijc.2910550118.
Recent data indicate a major role for IL-10 in suppressing immune and inflammatory reactions. To date, expression of human IL-10 has been attributed primarily to helper T lymphocytes, activated monocytes, and neoplastic B cells, and was often found to be associated with IL-6 expression. In this study we sought to determine whether non-hematopoietic human tumor cell lines produce IL-10 and, if so, what is the relationship between IL-10 and IL-6. Using ELISA, we determined IL-10 and IL-6 levels in culture supernatants of 48 cell lines established from carcinomas of the kidney, colon, breast and pancreas, malignant melanomas and neuroblastomas. IL-6 protein was secreted by 28 of the tumor cell lines; IL-10 was measurable in 15 cell lines. IL-6 secretion was maximal and most frequent in renal-cancer cell lines, while IL-10 production was found to be highest and most common among cell lines derived from colon carcinomas. IL-10 in conditioned medium of one of the colon carcinoma cell lines (CCL222) was bio-active, as demonstrated in the mouse MC/9 mast-cell-line assay and in human mixed-lymphocyte reactions. In both assays, IL-10 bio-activity was neutralized by an anti-IL-10 monoclonal antibody. Expression of IL-6 and IL-10 was confirmed by RNA analysis using message amplification by PCR and sequencing of amplified cDNA. LPS, IL-1 alpha, and TNF-alpha strongly enhanced the release of IL-6 by RCC cells, but only marginally affected IL-10 production in colon-carcinoma cells. IL-10 secretion by colon-carcinoma cells was moderately stimulated by IFN-gamma and IL-4. Dexamethasone suppressed the release of IL-6, but had no inhibitory effect on IL-10 secretion. Our results demonstrate that tumor cell lines established from certain types of human carcinomas are capable of expressing and releasing IL-6 and/or IL-10, suggesting a role of these cytokines in solid-tumor development and anti-tumor immunity.
近期数据表明白细胞介素-10(IL-10)在抑制免疫和炎症反应中起主要作用。迄今为止,人类IL-10的表达主要归因于辅助性T淋巴细胞、活化的单核细胞和肿瘤性B细胞,并且常常发现其与IL-6的表达相关。在本研究中,我们试图确定非造血性人类肿瘤细胞系是否产生IL-10,如果产生,那么IL-10与IL-6之间的关系是什么。使用酶联免疫吸附测定(ELISA),我们测定了从肾癌、结肠癌、乳腺癌和胰腺癌、恶性黑色素瘤及神经母细胞瘤建立的48个细胞系的培养上清液中的IL-10和IL-6水平。28个肿瘤细胞系分泌IL-6蛋白;15个细胞系可检测到IL-10。IL-6的分泌在肾癌细胞系中最高且最常见,而IL-10的产生在源自结肠癌细胞系中最高且最普遍。在小鼠MC/9肥大细胞系测定和人类混合淋巴细胞反应中证实,其中一个结肠癌细胞系(CCL222)条件培养基中的IL-10具有生物活性。在这两种测定中,IL-10的生物活性均被抗IL-10单克隆抗体中和。使用聚合酶链反应(PCR)进行信息扩增并对扩增的互补DNA(cDNA)进行测序,通过RNA分析证实了IL-6和IL-10的表达。脂多糖(LPS)、白细胞介素-1α(IL-1α)和肿瘤坏死因子-α(TNF-α)强烈增强肾癌细胞释放IL-6,但仅轻微影响结肠癌细胞中IL-10的产生。γ-干扰素(IFN-γ)和白细胞介素-4(IL-4)适度刺激结肠癌细胞分泌IL-10。地塞米松抑制IL-6的释放,但对IL-10的分泌没有抑制作用。我们的结果表明,从某些类型的人类癌建立的肿瘤细胞系能够表达和释放IL-6和/或IL-10,提示这些细胞因子在实体瘤发展和抗肿瘤免疫中起作用。