Hepal-6 RNA脉冲BMDCs瘤苗抗小鼠HCC免疫效应的研究

目的观察Hepal-6RNA脉冲BMDCs瘤苗能否激发有效的机体抗肿瘤免疫反应。方法用Hepal-6RNA脉冲骨髓分离培养5d的BMDCs,瘤苗的抗肿瘤效应通过C57BL/6J小鼠肝细胞癌(HCC)模型验证,即C57BL/6J小鼠皮下接种Hepal-6细胞8d后,成瘤小鼠分为2组:对照组(注射无血清RPMI-1640)和实验组(足垫部注射Hepal-6RNA脉冲BMDCs瘤苗),30d后处死小鼠并取淋巴结、脾和瘤体冰冻切片,进行CD4、CD8、CD25和Foxp3免疫组织化学染色。结果免疫组织化学检测显示,淋巴结、脾、肿瘤内有大量CD4+T细胞、CD8+T细胞、CD25+T细胞和Foxp3+Tregs细胞浸润。与对照组小鼠比较,足垫部注射Hepal-6RNA脉冲BMDCs瘤苗的实验组小鼠淋巴结、脾、瘤内CD4+T细胞和CD8+T细胞浸润显著增加,CD25+T细胞和Foxp3+Tregs浸润明显减少。结论 Hepal-6RNA脉冲BMDCs瘤苗能下调小鼠HCC瘤内CD25+T细胞和Foxp3+Tregs浸润,促进CD4+T细胞和CD8+T细胞的增殖和活化,增强D4+T细胞、CD8+T细胞归巢至淋巴结和脾,具有抗瘤免疫效应。

Antitumor immune efficacy of Hepal-6 RNA Pulsed BMDCs in mouse hepatocellular carcinoma

Objective To test whether mouse bone marrow derived dendritic cells （BMDCs） pulsed with Hepal-6 RNA can induce immune responses against tumor. Methods The efficacy of this vaccine was evaluated using a mouse HCC model. Hepal-6 cells were injected subcutaneously into the left flank of C57BL/6J mice to inoculate tumors. Eight days later, tumor-bearing mice were divided into 2 groups. controlled tumor-bearing mice injected with serum-free RPMI-1640; tested tumor-bearing mice injected with Hepal-6 RNA pulsed BMDCs into their hind footpads. 30 days later, all mice were killed for frozen sections of lymph nodes, spleens and tumors. IHC stained and analyzed the infiltration of CD4＋T, CD8＋ T, CD25＋ T cells and Foxp3＋ Tregs in those organs. Results Mice injected with Hepal-6 RNA pulsed BM- DCs showed obviously more CD4＋ T and CD8＋ T cells but fewer CD25＋ T cells and Foxp3＋ Tregs infiltra- ted into lymph nodes, spleens and tumor sites, when compared with serum-free RPMI-1640 mice. Conclu- sion The vaccine of Hepal-6 RAN pulsed BMDCs can downregulate the infiltration of CD25＋ T cells and Foxp3＋ Tregs, promote the proliferation and activation of CD4＋ T and CD8＋ T cells in mouse HCC, and enhance the homing of CD4＋ T and CD8＋ T cells in lymph nodes and spleens.