HSP70多肽复合物修饰DCs疫苗抗胰腺癌荷瘤小鼠的实验研究  

Antitumor effect of DCs vaccine modified by HSP70 peptide complex purified from mouse pancreatic carcinoma cells(MPC-83) in vivo

作　　者：张晓娟[1] 董坚[1] 吴振林[1] 洪敏[1] 邢海霞[1] 

机构地区：[1]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》2009年第9期792-796,共5页Chinese Journal of Immunology

基　　金：云南省科技条件平台建设项目(No.2007DA006)资助

摘　　要：目的:探讨负载热休克蛋白70多肽复合物(HSP70-PC)抗原的树突状细胞(DCs)疫苗对胰腺癌荷瘤小鼠的免疫治疗作用。方法:采用低渗裂解、ConA-Sepharose亲和层析及ADP-Agarose亲和层析法,从小鼠胰腺癌(MPC83)肿瘤组织中纯化HSP70-PC,修饰小鼠骨髓来源的树突状细胞(DCs),制备树突状细胞HSP70多肽肿瘤疫苗,用MTT法检测混合淋巴细胞反应(MLR)中HSP70-PC致敏DC对CTL的增殖及活化效果;建立MPC83荷瘤小鼠模型,观察树突状细胞HSP70多肽肿瘤疫苗对荷瘤小鼠治疗的效果和小鼠存活期。结果:经上述方法分离、纯化获得了较高纯度的HSP70-PC蛋白质;HSP70-PC在1.5～2.0μg/ml范围内可达到刺激树突状细胞最强效果,用HSP70-PC修饰的DCs能增强T细胞的增殖和活化能力;应用树突状细胞HSP70多肽肿瘤疫苗治疗荷瘤小鼠能显著抑制荷瘤小鼠肿瘤的生长,延长荷瘤小鼠存活期。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,其修饰的DCs疫苗用于荷瘤小鼠免疫治疗有显著疗效,为临床胰腺癌生物免疫治疗奠定基础。Objective：To investigate the effects of dendritic cells （DCs） vaccine loading with the heat shock protein 70 peptide complex（HSP70-pc） antigens against pancreatic cancer in the tumor-beating mice as the regimen of immune treatment. Methods： HSP70-PC was purified from the tumormass of pancreas cancer cell line （MPC83）-beariog mouse by ConA-Sepharose and ADP-Agarose afiqnity chromatography. Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL-4. MTT method was applied to analyze the ability of DCs, to induce preliferation of auto-lymphocytes by mixed lymphocyte reaction （MLR）. MPC83 tumor-bearing model of mice was established, which were treated by using dendritic cells pulsed with HSP70 peptide vaccine. The tumor size and survival of the mice were observed. Results ： Following the above method of separation and purification to obtain a higher purity of HSP70-PC protein; HSP70-PC at 1.5-2.0 μg/ml could be achieved the strongest effect to stimulate the dendritic cells. DCs modified with HSP70-PC could enhance the capacity of proliferation and activation for T cell in MLR. Dendritic cell vaccine pulsed with HSP70 peptide could significantly inhibit tumor growth in vivo and prolong survival time of tumor-bearing mice. Conclusion： High purity HSP70-PC for clinically individualized immunotherapy can be extracted from pancreatic cancer by using hypobarie affinity chromatography. DC vaccine modified with HSP70-PC can effectively induce antitumor immune response. This outcome lay the foundation for the clinical bio-immune therapy of pancreatic cancer.

关 键 词：胰腺癌 热休克蛋白70多肽复合物 树突状细胞 肿瘤免疫 

分 类 号：R730.51[医药卫生—肿瘤]