抗人P185^erbB2 scFv-Fc-IL-2增强LAK样和ADCC杀伤作用及机制的实验研究  

Experimental study on the enhancement of killing effect of LAK-like cells and ADCC induced by anti-human P185^erbB2 scFv-Fc-IL-2 fusion protein and its mechanisms

作　　者：王军[1] 张玲[1] 毛海婷[1] 郭宁[2] 施明[2] 沈倍奋[2] 顾洪涛[1] 李翠玲[1] 

机构地区：[1]山东省医学科学院基础医学研究所 山东省医药卫生肿瘤免疫与中药免疫重点实验室 山东省现代医用药物与技术重点实验室,济南250062 [2]军事医学科学院基础医学研究所分子免疫重点实验室

出　　处：《中华微生物学和免疫学杂志》2008年第2期107-111,共5页Chinese Journal of Microbiology and Immunology

基　　金：山东省科技攻关重点项目资助（No.2004GG2202149）

摘　　要：目的将抗人P185^erbB2 scFv-Fc-IL-2融合蛋白（HFI）分别作用于表达高、中、低3个水平erbB2受体的SKOV3、MCF-7、SGC-7901三株肿瘤细胞和健康人外周血单个核细胞（PBMC），探讨HFI调变肿瘤细胞表面分子，激活免疫效应细胞的机制；模拟体内肿瘤组织，将Hn-PBMC-肿瘤细胞混合培养，探讨HFI对分别表达高、中、低3个水平erbB2肿瘤细胞的淋巴因子激活的杀伤细胞（LAK）样和抗体依赖性细胞介导的细胞毒（ADCC）作用，为临床应用提供实验依据。方法MTT法检测细胞增殖、杀伤活性；流式细胞术检测细胞表面分子的表达变化；应用非核素细胞毒试剂盒观察HFI介导ADCC杀伤作用。结果HFI处理后SKOV3细胞表面杀伤相关分子细胞间黏附分子-1（ICAM-1）、Fas表达水平分别由24．85％、0．53％增高到85．36％、59．19％；SKOV3、MCF-7、SGC-7901三株肿瘤细胞erbB2表达水平分别由98．48％、42．60％、36．66％下降到94．01％、30．95％、12．36％。HFI刺激后人PBMC的增殖活性显著增强，呈时间依赖效应，CD3^＋CD8^＋T细胞和CD3^-CD16^＋CD56^＋NK细胞分别由24．37％、6．90％提高到38．80％、13．45％，CD25、淋巴细胞功能抗原-1（LFA-1）、FasL表达水平分别由3．99％、86．52％、5．02％提高到12．96％、99．06％、16．19％。HFI活化的人PBMC对不同erbB2表达水平肿瘤细胞的LAK样杀伤活性，在各效靶比均比对照组明显提高。HFI能够介导和增强人PBMC对表达erbB2肿瘤细胞的ADCC杀伤作用。结论HFI上调SKOV3细胞表面杀伤相关分子ICAM-1、Fas表达，下调肿瘤细胞表面erbB2表达。HFI对人PBMC具有明显的激活增殖作用，活化的人PBMC对表达不同水平erbB2肿瘤细胞的LAK样杀伤作用均显著增强。HFI能够介导和增强人PBMC对肿瘤细胞的ADCC杀伤作用，且杀伤活性的高低与肿瘤细胞表面erbB2表达水平的高低呈平行关系。Objective To explore the mechanism by which the anti-human P185^erbB2scFv-Fc-IL-2 （HFI） modulates tumor surface molecules and activates immune effector cells in vitro. Methods MTT assay was used to test the proliferation and the LAK-like cytotoxicity. Flow cytometry assay was used to test the expression of ICAM-1, Fas and erbB2 receptors in tumor cells and the expression levels of CD molecules FasL and LFA-1 in human PBMC. Antibody-dependent cell-mediated cytotoxicity （ADCC） mediated by HFI against SKOV3, MCF-7 and SGC-7901 tumor cells was explored by LDH release assay. Results The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI were upregulated, from 24.85% and 0.53% to 85.36% and 59.19% respectively, while the expression levels of erbB2 on SKOV3, MCF-7 and SC, C-7901 tumor cells treated with HFI were downregulatod, from 98.48%, 42.60% and 36.66% to 94.01% ,30.95% and 12.36% respectively. HFI could significantly enhance the proliferation activity of human PBMC, and CD3^＋ CD8^＋ T cells and CD3^- CD16^＋ CD56^＋ NK cells were elevated, from 24.37% and 6.90% to 38.80% and 13.45% respectively. The expression levels of CD25, LFA-1 and FasL were significantly enhanced from 3.99%, 86.52% and 5.02% to 12.96%, 99.06% and 16. 19%. The LAK-like cytotoxicity of human PBMC treated with HFI against SKOV3, MCF-7, SGC-7901 tumor cells was significantly improved; HFI was effective in mediating ADCC against SKOV3, MCF-7 and SGC-7901 tumor cells which expressed high, medium and low levels of erbB2, respectively, and HFI-induced ADCC was correlated with the degrees of erbB2 expression on the tumor cells. Conclusion The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI are significantly upregulated. The expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI are downregulated. HFI can significantly enhance the proliferation activity of human PBMC. The LAK-like cytotoxicity of human PBMC treated with HFI against tumor cells is significantly enhanced. HFI i

关 键 词：ERBB2 HFI融合蛋白 免疫细胞因子 细胞毒性 抗肿瘤 

分 类 号：R686[医药卫生—骨科学]