小鼠肝癌全细胞抗原致敏骨髓DC激活TIL抗癌机制的研究  

Antitumor mechanism of the tumor-infiltrating lymphocyte stimulated by mice bone marrow DCs pulsed with mice hepatoma H22 full-cell antigen

作　　者：张志明[1] 冯钟煦[1] 周敏[1] 刘剑勇[1] 赵荫农[1] 张春燕[1] 唐凯[1] 吕丽琼[1] 罗善超[1] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,广西南宁530021

出　　处：《中华肿瘤防治杂志》2013年第13期977-981,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：广西医疗卫生重点科研课题(重200611);广西科学研究与技术开发计划(桂科攻0719006-2-5);广西科学基金(桂科自0728196)

摘　　要：目的:通过分析小鼠肝癌H22细胞全细胞性抗原致敏小鼠骨髓树突状细胞(DC)前后DC表型变化及其分泌细胞因子的变化,探讨肝癌细胞全细胞性抗原致敏DC激活肿瘤浸润性淋巴细胞(TIL)的抗癌机制。方法:取得小鼠骨髓细胞并诱导生成DC,用冻融法制备的小鼠肝癌H22细胞全细胞抗原致敏,然后用已致敏的DC激活TILs,测定DC致敏前后DC表面抗原CD80、CD86、CD40和MHCⅡ表达变化及DC分泌细胞因子IL-12、IL-2、IFN-γ和TNF-α的水平,评估激活前后TIL对H22细胞的杀伤活性,同时以小鼠脾淋巴细胞作为杀伤对照。结果:致敏后小鼠骨髓DC表面抗原CD80、CD86、CD40和MHCⅡ表达率分别为(57.55±7.32)%、(54.49±14.20)%、(46.79±8.25)%和(53.94±13.94)%,明显高于未致敏DC的(20.01±5.22)%、(24.56±9.08)%、(18.06±5.13)%和(30.24±8.39)%,P值均<0.05;DC上清液中IL-12、IL-2、IFN-γ和TNF-α浓度分别为(80.40±1.33)、(94.67±3.36)、(29.83±1.20)和(75.01±4.10)ρg/mL,明显高于未致敏DC的(19.35±0.99)、(11.25±0.50)、(1.05±0.09)和(2.02±0.27)ρg/mL,P值均为0.000。经致敏后成熟DC激活的TIL对H22细胞杀伤率为(81.80±2.90)%,明显高于未激活TIL、激活或未激活小鼠脾淋巴细胞的(62.64±3.94)%、(47.35±3.40)%和(28.45±2.56)%。结论:H22细胞全细胞抗原致敏DC后,其表型变化及细胞因子分泌增多是其诱导活化TIL抗癌活性的可能机制。OBJECTIVE： To study the anti-mouse hepatoma effect of tumordnfihrating lymphocytes （TILs） stimula ted by dendritic cells （DCs） pulsed with the tumor full-cell antigen in vitro. METHODS： DCs were isolated from BALB/c mouse bone marrow and stimulated by granulocyte macrophage colony stimulating factor （GM-CSF）, interleukin-4 （II.-4） and H22 full-cell antigen. The maturation surface markers CD80,CD86,CD40, MHC ]] were detected by flow cytometry before and after pulsing with mice hepatoma lysates. The cytokines IL-12, IL-2, IFN-7, TNF-a secreted by DC were detec- ted by the enzyme-linked immuno sorbent assay. The cytotoxic potency of the TILs was assessed both stimulated and uns- timulated,and taking the spleen lymphocytes as control groups. RESULTS： After sensitization CD80, CD86, CD40, MHC [I of DCs surface antigen expression rates were （57.55 ±7.32）%, （54.49 ±14.20）%, （46.79 ± 8.25）%and （53.94 ± 13.94 ）%, which was significantly higher than those of non sensitized DCs （ 20.01 ± 5.22） %, （ 24.56 ± 9.08） %, （ 18.06 ± 5.13） %, （30. 24 ± 8. 39）% （P〈0. 05）% DCs supernatant IL-12, IL-2, IFN-7, TNF-a concentration respectively were （80.40±1.33） ,（94.67±3.36）,（29.83±1.20） and （75.01±4. 10） tog/mL,which was significantly higher than that in the unsensitized DCs （ 19.35 ±0.99）, （ 11.25 ± 0.50）, （ 1.05 ±0.09）, （ 2.02 ± 0.27） tog/mL （P = 0. 000）. TILs stimulated by the DCs on H22 whole-cell antigen had very special efficient in antitumor effect on the H22 cell in vitro and the killingrate was （81.90 ± 2.90）% which was much higher than those of TILs not stimulated by DCs and splenocytes stimulated by DCs as well as splenocytes not stimulated by DCs. H22 cell killing rates were （62.64±3.94） % , （47.35 ±3.40） % and （28.45±2.56） % ,respectively. CONCLUSION： DCs phenotypic changes sensitized by H22 cell antigen and cytokine se- cretion promotion are the possible mechanisms in inducing and activating

关 键 词：肝肿瘤 全细胞抗原 树突细胞 肿瘤浸润性淋巴细胞 抗癌机制 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]