HSP70多肽复合物修饰树突细胞抗胰腺癌研究  

Induction specific anti-tumoral immunity against pancreatic cancer by dendritic cells modified by HSP70 peptide complex from pancreatic cancer cells

作　　者：董坚[1] 吴振林[1] 贾伟[1] 张晓娟[1] 陈明清[1] 

机构地区：[1]云南省昆明医学院第一附属医院生物治疗科,昆明650032

出　　处：《中国免疫学杂志》2008年第1期28-33,共6页Chinese Journal of Immunology

基　　金：昆明医学院创新群体基金(No.KMC2005DG01)资助

摘　　要：目的:研究肿瘤热休克蛋白70(HSP70)多肽复合物修饰树突状细胞激活淋巴细胞治疗胰腺癌的策略和方法。方法:采用低渗裂解,ConA-Sepharose亲和层析柱及ADP-Agarose亲和层析柱,从小鼠胰腺癌(MPC83)瘤块中纯化HSP70多肽复合物;纯化出的70KD蛋白修饰小鼠骨髓来源诱导树突细胞(DC)并制备树突细胞HSP70多肽肿瘤疫苗,MTT法检测修饰后DC增殖活性;用修饰后DC激活小鼠脾淋巴细胞,MTT法检测激活淋巴细胞在不同效靶比下对MPC83的体外杀伤活性。结果:获得较高纯度分子量为70kD左右的蛋白质;50～100ngHSP70多肽复合物可修饰104树突细胞,每克瘤块能获取HSP70多肽复合物约100μg;来自MPC83细胞瘤块HSP70多肽复合物激活的淋巴细胞能特异性杀伤MPC83细胞。结论:采用低压亲和层析柱可从胰腺癌瘤块中获得较高纯度HSP70多肽复合物,HSP多肽复合物DC疫苗用于胰腺癌细胞免疫治疗能获得体外杀伤效果,为临床胰腺癌生物免疫治疗奠定基础。Objective：To investigate the strategies and methods of pancreatic cancer immunotherapy adopt cytotoxic T lymphocytes activated by dendritic cells （DCs）, which modified by heat shock protein 70 peptide complex （HSP70-PC）. Methods：HSP70 peptide complex from lumps of tumor-bearing mouse innoculated with pancreas cancer cell （ MPC83 ） were purified by ConA-Sepharose and ADP-Agarose affinity chromatography. Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL- 4. MTF method was applied to analyses the proliferation activities of DCs. To achieve the specific T lymphocyte cells, DCs modified with HSP70 peptide complex were coculture with murine spleen lymphocytes for 48 hours. Then these activated lymphocytes were cocultared with MPC83 at ratio of effect-target 10：1 and 20：1 in vitro and the killing activities were detected by MTF method. Results：High purify HSP70-PC was obtained; 104DCs need 50 - 100ng of HSP70-PC to modify, 100μg of HSP70-PC peptide complex can be extracted from per lg lump; T cells stimulated by HSP70-PC modify DCs showed more specific cytotoxic activity to MPC83 than lymphocytes activated by DCs without HSP70-PC in vitro. Conclusion：We can extract high purity HSP70-PC for clinical individual immunotherapy from pancreatic cancer lump by using hypobaric affinity chromatography; DCs vaccine modified with HSP70-PC has significant kill effect against pancreatic cancer cell in vitro.

关 键 词：胰腺癌 热休克蛋白70多肽复合物 蛋白质纯化 树突状细胞 

分 类 号：R730.54[医药卫生—肿瘤]