抗人P185^(erbB2) scFv-Fc-IL-2调变肿瘤细胞表面分子和激活免疫效应细胞  

Anti-P185^(erbB2) scFv-Fc-IL-2 modulates tumor cell surface molecules and activates immune effector cells

作　　者：王军[1] 张玲[1] 顾洪涛[1] 郭宁[2] 施明[2] 沈倍奋[2] 毛海婷[1] 李翠玲[1] 温培娥[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,北京100850

出　　处：《中国肿瘤生物治疗杂志》2006年第6期412-416,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省科技攻关基金资助项目(No.2004GG2202149)

摘　　要：目的将抗人P185erbB2scFv-Fc-IL-2融合蛋白(HFI)作用于人卵巢癌细胞株SKOV3细胞和人外周血单个核细胞(PBMC),通过体外实验阐明HFI调变肿瘤细胞表面分子和激活免疫效应细胞的抗肿瘤机制,为HFI临床应用提供实验依据。方法MTT法检测细胞增殖、杀伤活性;流式细胞术观察细胞表面分子的表达水平;生物活性法检测细胞膜相关TNF杀伤活性;RT-PCR检测细胞穿孔素表达水平。结果HFI处理后,未观察到对SKOV3细胞增殖活性的直接抑制作用;SKOV3细胞表面杀伤相关分子ICAM-1、Fas表达率分别由24.85%、0.53%增高到85.36%、59.19%(P<0.01);人PBMC的增殖活性增强,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%(P<0.01);CD25、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%(P<0.01);穿孔素基因、膜相关TNF均表达增强,LAK样、NK样杀伤活性在各效靶比时均明显增高(P<0.01)。结论HFI提高SKOV3细胞杀伤相关分子ICAM-1、Fas表达水平,并且对人PBMC有明显的增殖活化作用,通过激活LFA-1/ICAM-1、Fas/FasL途径提高杀伤介质穿孔素和膜相关TNF的释放,增强LAK样、NK样杀伤活性。Objective： To elucidate the mechanisms by which anti-P185erbB2scFv-Fc-IL-2 （HFI） modulates tumor surface molecules and activates immune effector cells in vitro. Methods： Ovarian Cancer Cell Line SKOV3 and human peripheral blood monocyte peripheral blood mononuclear cells （PBMC） were treated with HFI. MTT assay was used to detect the proliferation and cytotoxicity; flow cytometry assay was used to examine expression of surface molecules; bioassay was used to examine cytotoxicity of membrane-associated TNF ; and perforin mRNA level was analyzed by RT-PCR assay. Results： HFI had no direct inhibitory effect on proliferation of SKOV3 cells. After HFI treatment, the expression levels of ICAM-1 and Fas on SKOV3 cells rose from 24.85% and 0.53% to 85.36% and 59.19%, respectively （P 〈0.01 ）. Besides, HFI significantly enhanced the proliferation of human PBMC. CD3^＋ CD8 ^＋T cells and CD3 CD16^＋ CD56 ^＋T cells rose from 24. 37% and 6.90% to 38.80% and 13.45% , respectively （P 〈 0.01 ）. Expression levels of CD25, LFA-1, and FasL significantly increased from 3.99%, 86.52%, and 5.02% to 12.96%, 99.06%, and 16.19%, respectivly （P 〈0.01 ）. Expression levels of Perforin and membrane-associated TNF were also up-regulated. Cytotoxicities of LAK and NK were both improved（P 〈0.01 ）. Conclusion： HFI can enhance the expression of ICAM-1 and Fas in SKOV3 cells and has obvious activating effect on PBMC. HFI can up-regulate the expression level of LFA-1/ICAM-1, Fas/FasL, promote the release of TNF and perforin, and improve LAK and NK cytotoxicity.

关 键 词：ERBB2 P185erbB2scFv—Fc—IL-2融合蛋白 卵巢癌 免疫细胞因子 杀伤活性 

分 类 号：R730.5[医药卫生—肿瘤]