CytoMPS原位疫苗治疗肝癌的抗瘤作用及其免疫效应  

Cytokines microparticles (CytoMPS)-in situ vaccination for hepatocellular carcinoma:in vivo and in vitro responses

作　　者：匡铭[1] 刁竞芳[2] 彭宝岗[1] 沈顺利[1] 王晔[1] 吴浩[3] 陈斌[1] 

机构地区：[1]中山大学附属第一医院肝胆外科,广州510080 [2]广东省中医院肝胆胰外科 [3]广州市第十二人民医院普外科

出　　处：《中华普通外科学文献（电子版）》2010年第5期10-12,36,共4页Chinese Archives of General Surgery(Electronic Edition)

基　　金：国家自然基金资助项目(30672052;30872486);广东省自然基金资助项目(8151008901000211)

摘　　要：目的制作成细胞因子缓释微粒原位瘤苗(CytoMPS-ISV),观测CytoMPS-ISV抗瘤作用及其免疫效应。方法 C57BL/6J小鼠皮下移植性肝癌模型瘤内注射CytoMPS3次制作CytoMPS-ISV,观察其抗肿瘤作用及小鼠生存率;流式细胞仪检测小鼠外周血淋巴细胞变化,免疫组化法检测肿瘤组织的CD4+、CD8+和NK1.1+细胞浸润情况,体外检测小鼠脾CTL和NK细胞的杀瘤活性,检测疫苗诱导脾CTL的抗体阻断作用。结果与PBS注射组和对照组相比,瘤内注射CytoMPS后肿瘤生长受到显著抑制(P<0.01)。瘤内注射CytoMPS组血液中及局部浸润的CD4+、CD8+和NK细胞明显高于PBS组和对照组(P<0.05);淋巴细胞杀伤试验结果显示瘤内注射CytoMPS组小鼠的脾CTL对靶细胞的杀伤率明显高于PBS组(P<0.01)和对照组(P<0.05);抗体阻断作用试验显示接种疫苗的小鼠脾CTL的杀瘤活性可被抗CD8+、抗MHC-I单克隆抗体所阻断,但不被抗CD4+、抗MHC-II单克隆抗体所阻断。结论 CytoMPS-ISV可以显著抑制肿瘤生长,能明显增强机体的抗肿瘤免疫作用,其诱导的CTL杀瘤特性是由MHC-I限制的CD8+T细胞所介导。Objective Toprepare the CytoMPS-related insitu vaccine（CytoMPS-ISV）and observeits protective and therapeutic antitumor effects in treatment of mice hepatoma.Methods Three times of intratumoral injection of CytoMPS in C57BL/6J mice subcutaneous hepatoma were performed, and the tumor growth and mice survival rates were observed.The proportions of peripheral blood lymphocytes were detected by FACS.The killing efficacy of cytotoxic T lymphocytes （CTLs） derived from spleens of CytoMPS-ISV vaccinated mice was evaluated with and without antibody-blocking preparation.Extent of tumor-infiltrating CD4＋、CD8＋ and NK1.1＋ cells were also detected with immunohistochemical stain （IHC）.Results Compared with the control group, the tumor growth in CytoMPS-ISV group was significantly inhibited （P〈0.01）.Moreover, the proportions of CD4＋, CD8＋ T cell and natural killer cell （NK） in peripheral blood in CytoMPS-ISV group were significantly higher, so were those of tumor-infiltrating lymphocytes in CytoMPS group.Compared with those in the control groups, the cytotoxicities of splenic CTL in CytoMPS-ISV group were significantly enhanced, which could be blocked by anti-mouse MHC Class I and anti-mouse CD8 antibody, but not by anti-mouse MHC Class II and anti-mouse CD4 antibody.Conclusion CytoMPS-ISV can significantly inhibit tumor growth and induce anti-tumor immunity.The killing efficacy of CytoMPS-ISV-induced CTLs is related to CD8＋ T cells restricted by MHC-I.

关 键 词：细胞因子缓释微粒 肝细胞癌 疫苗 免疫治疗 

分 类 号：R735.7[医药卫生—肿瘤]