摘要
目的观察人体 CIK(cytokine-induced killer)细胞的体外增殖力及杀伤肝癌细胞活性。方法采取健康自愿者的成分血,用淋巴细胞分离液分离获得外周血单个核细胞(PBMC),在体外培养条件下,通过加入不同细胞因子(如 IFN-γ,IL-1,IL-2,CD3 mAb)培养诱导成 CIK 细胞和 LAK 细胞,观察不同培养细胞的增殖能力;用流式细胞仪对培养细胞做细胞表型动态分析,观察培养细胞的表面标志;与 LAK 细胞作对比,用 MTT 法测定并比较不同效应细胞的体外抗肝癌细胞活性。结果 CIK 细胞在培养14d 后开始大量增殖且在含人血清的培养基中增殖数量最多,可达500多倍,经表型分析表明,CIK 细胞属于异质细胞群,在培养过程中,CD^(3+)CD^(56+)双阳性细胞的绝对数量获得了大量增殖,至56d 时可占细胞总数的51.26%,是 CIK 主要的效应细胞;实验表明,CIK 细胞体外杀伤肝癌细胞的活性明显优于 LAK 细胞。结论 CIK 细胞是一种具有较强杀伤肝癌细胞的免疫活性细胞,有可能应用于抗肿瘤的生物治疗。
AIM To evaluate the proliferation profile and the antitumor cytotoxicity of human cytokine-induced killer (CIK)cells in vitro. METHODS Peripheral blood mononuclear cells(PBMC) were obtained from healthy volunteers by Ficoll gradient centrifugation and CIK cells were induced by culturing PBMC in the completed medium containing some cytokines such as IFN-γ,IL-1,IL-2 and mAb against CD3;the expanding numbers of cells were investigated;CIK cells were analyzed at different intervals by flow cytometric analyser to find out the phenotypic characterization and identification of effector cells;compared with LAK cells, the cytotoxicity of CIK cells against hepatocellular carcinoma cells was determined by MTT assay. RESULTS CIK cells increased significantly after 14 days of culture and expanded more than 500-fold in the medium consisting of 100 mL.L^(-1) human AB serum.The data of flow cytometric analysis showed that CIK cells was a heterogeneous population of effector cells.During cell generation,there was a steady increase in both the absolute number and the percentage of CD^(3+) CD^(56) cells, reaching 51.26% on day 56 of culture.CD^(3+) CD^(56+) T cells were the major cytolytic effectors.CIK cells possessed a higher proliferation rate and higher antitumor cytotoxic activity in vitro than LAK cells. CONCLUSION CIK cells were highly efficient cytotoxic effector cells against hepatocellular carcinoma cells and may be used for adoptive antitumor immunotherapyo
出处
《世界华人消化杂志》
CAS
2000年第8期863-866,共4页
World Chinese Journal of Digestology
关键词
肝肿瘤
免疫活性
CIK细胞
外周血单个核细胞
Subject headings liver neoplasms
immunocompetence
cytokine-induced killer cells
peripheral blood mononuclear cells
lymphokineactivated killer
tumor-infiltrating lymphocytes
