肝癌细胞对重组人HSP70联合肝癌组织冻融抗原修饰树突状细胞的免疫应答  

Dendritic cells pulsed with recombinant human HSP70 and freeze-thaw cell lysates derived from HepG2 cells induce specific anti-tumor immune responses

作　　者：魏云涛[1] 徐阳[1] 姜晓峰[1] 郭大伟[1] 王学范[1] 梁健[1] 

机构地区：[1]中国医科大学附属第四医院肝胆外科,辽宁沈阳110032

出　　处：《中华肿瘤防治杂志》2009年第8期573-577,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省教育厅2008高等学校科研项目计划(2008824)

摘　　要：目的:研究重组人热休克蛋白70(rhHSP70)联合肝癌组织冻融抗原修饰的树突状细胞(dendriti ccell,DC)诱导对肝癌细胞的免疫杀伤效应。方法:外周血单个核细胞经粒-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素4(IL-4)诱导生成DC,负载冻融抗原的同时加入rhHSP70,不同分组致敏的DC激活淋巴细胞生成肿瘤抗原特异性细胞毒性T淋巴细胞(cyto-toxic T cells,CTL),四甲基偶氮唑蓝(MTT)法及3H-TdR法检测DC刺激淋巴细胞增殖能力,MTT法检测CTL对肝癌细胞的体外杀伤活性,酶联免疫吸附试验(ELISA)测定细胞因子的分泌,流式细胞术(FCM)检测DC表型变化。结果:冻融抗原致敏的DC可明显促进淋巴细胞增殖,能有效呈递肝癌冻融抗原,诱导产生抗原特异性CTL,联合rhHSP70能进一步增强CTL对肝癌细胞的杀伤作用。结论:肝癌冻融抗原联合rhHSP70修饰的DC诱导CTL对肝癌细胞能产生高效杀伤作用。OBJECTIVE：To investigate the anti-tumor effect of dendritic cells（DC） pulsed with recombinant human HSP70 and freeze-thaw cell lysates derived from HepG2 cells. METHODS： DC were derived from peripheral blood mononuclear cells of healthy donors and then stimulated in vitro with a granulocyte-maerophage colony-stimulating factor（GM-CSF） and interleukin-4 （IL-4）. DC were loaded with freeze-thaw lysate derived from HepG2 cells, and at the same time rhHSP70 as an immunol adjuvant was added. The different groups each induced proliferation of tumor specific cytotoxic T cells （CTL） in the presence of human GM-CSF and IL-4. The proliferation of lymphocytes was measured by MTT assay and 3 H-TdR assay. The anti tumor effect activated by the vaccine of DC was measured by MTT assay. The cytokine release of DC and lymphocytes were detected by ELISA. DC phenotype was observed by FCM. RESULTS： DC pulsed with freeze-thaw cell lysates derived from HepG2 cells enhanced the growth expansion of lymphocytes, and presented effectively tumor antigen to stimulate generation of specific CTL. With the addition of rhHSP70, this activity was further enhanced. CONCLUSION： DC pulsed with rhHSP70 and freeze-thaw cell lysats derived from HepG2 cells are very effective and in activating specific T-cell responses against HepG2 cells in vitro.

关 键 词：肝肿瘤/免疫学 热休克蛋白质70 树突细胞 T淋巴细胞 细胞毒性 

分 类 号：R735.7[医药卫生—肿瘤]