微波消融灭瘤联合瘤内接种树突状细胞诱导特异性抗肝癌免疫

Microwave ablation combined with intra-tumoral inoculation of dendritic cells induces specific immunity against hepatocellular carcinoma

目的探讨微波消融（MWA）灭瘤联合瘤内接种帝寸突状细胞（DCs）诱导特异性抗肝癌免疫的效能。方法采用GMCSF联合IL-4体外培养C57BL／6小鼠骨髓来源的DCs。丁第6天收集使用。建立C57BL／6小鼠皮FHepal-6肝癌模型，随机分为对照组、瘤内接种DCs组（DC组）、肿瘤微波消融组（MWA组）及8中瘤微波消融＋智内接种DCs组（MWA＋DC组）。免疫组织化学法检测肿瘤组织内CD4＋和CD8T细胞的浸润．MTT法检测小鼠脾脏细胞对Hepa16的特异性杀伤活性，观测各组小鼠肿瘤生K情况。结果免疫组织化学法检测显示MwA＋DC组肿瘤组织内有大量的CD4-和CD8＋T淋巴细胞浸润，显著高于其它组（P〈0．05）。MWA＋DC组脾细胞对Hepal-6细胞有特异性杀伤效能，在E／T=40和100时。MWA＋DC组脾细胞对Hepal-6细胞的特异性杀伤力显著高于对照组、DC组及MWA组（P〈0．05）。MWA＋DC组小鼠肿瘤完全消退率显著高于其它各组（P〈0．05）。结论MWA联合瘤内接种DCs可有效诱导机体产生特异性抗肝癌免疫．是预防MwA治疗后肝瘤复发的一种有效方法。

Objective To investigate the efficacy of microwave ablation combined with intra-tu- moral inoculation of dendritic cells （DCs） in inducing specific immunity against hepatoeellular carcino- ma （HCC） in vivo. Methods C57BL/6 bone marrow derived DCs were grown in vitro for 6 d with use of murine recombinant GM-CSF and IL-4. C57BL/6 mice were inoculated subcutaneously with Hepal-6 cells to establish subcutaneous HCC. and randomized into control group, group receiving intratumoral inoculation of DCs （DC group）, group receiving microwave ablation of tumor （MWA group）, and group receiving microwave ablation of tumor plus intra tumoral injection of I）Cs （MWA＋ DC group）. Immunohistochemical staining was employed to evaluate intra tumoral infiltration of CD4＋ and CD8＋ lymphocytes. Cytotoxicity of splenocytes against Hepal-6 was ewduated using MTT method. Tumor growth in each group was compared. Results Immunohistoehemical staining illustra ted that MWA＋DCs group had a markedly greater nmnber of CD4＋ and CD8＋ T lymphocytes inside tumor tissue as compared with the control group, DC groupand MWA group （P〈0.05）. Splenocytes from MWA＋DCs group had a remarkably higher specific cytotoxicily against Hepal 6 at E/T ratio of 40 and 100 than those from control group, DC group and MWA group （P〈0.05）. Tumor regression rate was significantly higher in MWA＋DCs group as compared with the control group, DC group and MWA group （P〈0.05）. Conclusion Intra-tumoral inoculation of DCs following MWA could effeetively elicit specific anti-tumoral immunity in vivo. It can be used as an effective approach for preven ting tumor recurrence after MWA.