Hepal-6细胞热休克后裂解蛋白致敏骨髓来源树突状细胞瘤苗的抗肿瘤免疫学效应  

Antitumor effect of bone marrow derived dendritic cells sensitized by heat shocked Hepal-6 cells lysated protein

作　　者：李日伦[1] 周爽[2] 秦杰[2] 张浩如[2] 梁春敏[2] 罗国容[3] 

机构地区：[1] 广西卫生职业技术学院 [2]复旦大学上海医学院解剖学与组织学胚胎学系,上海200032 [3]广西医科大学组织学与胚胎学教研室,南宁530021

出　　处：《解剖学杂志》2014年第2期137-141,F0002,共6页Chinese Journal of Anatomy

基　　金：国家自然科学基金青年基金项目（30500280）和面上项目（30871312）

摘　　要：目的：研究Hepal-6细胞热休克后裂解蛋白致敏的骨髓来源树突状细胞（BMDCs）的抗肿瘤免疫生物学效应.方法：用Hepal-6细胞热休克后裂解蛋白致敏BMDCs制成瘤苗,瘤苗的效应通过C57BL/6J小鼠肝细胞癌模型验证.皮下接种Hepal-6细胞8d后,成瘤小鼠随机分实验组,足垫部皮下注射Hepal-6细胞热休克后裂解蛋白致敏BMDCs;无血清RPMI-1640注射组、BMDCs注射组和单纯Hepal-6细胞裂解蛋白致敏BMDCs注射组.取肿瘤组织分离成细胞悬液,FACS检测CD11c＋细胞、CCR7＋细胞、CD80＋细胞和CD86＋细胞;取淋巴结、脾、肿瘤冷冻切片并采用免疫组织化学检测淋巴结、脾、肿瘤内Foxp3＋细胞浸润的改变;ELISA检测瘤内转化生长因子-p1（TGF-β1）、白细胞介素-10（IL-10）、γ干扰素（IFN-γ）、白细胞介素-2（IL-2）的表达.结果：实验组肿瘤组织比各对照组有更多的CD11c＋细胞、CCR7＋细胞、CD80＋细胞和CD86＋细胞浸润;与各组对照相比较,实验组Foxp3＋细胞在淋巴结、脾、肿瘤内浸润明显减少;实验组细胞因子TGF-β1、IL-10低于各对照组,IFN-γ、IL-2则高于各对照组.结论：Hepal-6细胞热休克后裂解蛋白致敏的BMDCs瘤苗能促进DCs在瘤内浸润和成熟,减少Foxp3＋细胞在肿瘤内的浸润,下调免疫抑制细胞因子TGF-β1、IL-10和上调免疫促进因子IFN-γ、IL-2分泌,有较强的抗肿瘤免疫生物学效应.Objective： The current study was conducted to test whether heat shocked Hepal-6 cells lysated protein sensitized bone marrow derived dendritic cells （BMDCs） can promote more effective antitumor biological activities. Methods： The efficacy of this vaccine was evaluated using a mouse hepatocellular carcinoma （HCC） model. Hepal-6 cells were injected subcutaneously into the left flank of C57BL/6J mice to inoculate tumors. After eight days, the tumor bearing mice were divided into four groups. （1） Test group： mice were injected with BMDCs sensitized by heat shocked Hepal-6 cell lysate proteins in hind footpads; （2） Three control groups.- in one group, mice were injected with serum-free RPMI-1640 in hind footpads; in another group, mice were injected with control BMI）Cs; in the third group, mice were injected with BMDCs sensitized by Hepal-6 cells lysates protein. Every 7 days for one injection, all two times. After 9-day treatment, all mice were sacrificed. FACS analysis for the infiltration of CDllc＋ , CCR7＋ , CDS0＋ and CD86＋ cells in tumor. IHC analysis for infiltration of Foxp3＋ Treg cells in the lymph nodes, spleens and tumor. The expression of TGF-β, IL-10, IL-2 and INF-γ in tumor sites was analyzed by ELISA assay. Results： The test group showed more CDllc＋ , CCR7＋ , CD80＋ and CD86＋ DC cells infiltrated into tumor sites. At the same time, this group also showed a notable decreasing expression of TGF-β, IL-10 and the obviously up-regulated expression of IL-2, INF-γ in tumor. It was also found that Foxp3＋ Treg cells showed a notable decrease in infiltrating into tumor sites when compared to the control mice. Conclusion： BMDCs vaccines sensitized by heat shocked Hepal-6 ceils lysates protein can promote a more functional antitumor effect and represent a promising approach toinduce immune-mediated regression of malignant tumor.

关 键 词：Hepal-6细胞热休克后裂解蛋白 骨髓来源树突状细胞 免疫调节性T细胞 

分 类 号：R512.6[医药卫生—内科学]