树突状细胞疫苗联合胸腺肽α1对人源化免疫重建裸鼠结肠癌生长的抑制效应  

In vivo study of DC-based immunotherapy combined with thymosiu-alphal on humanized nude mice with colon cancer

作　　者：马云龙[1] 郑铮[1] 郭华[1] 李保东[1] 谢绍建[2] 李桂馨[2] 刘津[1] 刘三光[3] 闫庆辉[1] 薛平[2] 蔡建辉[1] 

机构地区：[1]河北医科大学第二医院胃肠外科,石家庄050000 [2]河北医科大学第二医院外科研究中心,石家庄050000 [3]河北医科大学第二医院肝胆外科,石家庄050000

出　　处：《中华实验外科杂志》2007年第11期1357-1359,共3页Chinese Journal of Experimental Surgery

基　　金：河北省科技厅博士基金(04547002D-9);河北省卫生厅重大攻关项目(2003001)

摘　　要：目的观察胸腺肽α1(Ta1)联合结肠癌细胞裂解物致敏树突状细胞(LyDCs)对人源化免疫重建裸鼠结肠癌的免疫治疗效应。方法常规DCs负载结肠癌细胞裂解物制备LyDCs疫苗,流式细胞仪(FCM)检测Tα1体外刺激前、后的LyDCs表型。HT-29结肠癌裸鼠模型成瘤后,经尾静脉注射人外周血T淋巴细胞6×10~6个/只,2d后,FCM检测裸鼠外周血人源性CD4^+、CD8^+ T细胞;将人源化免疫重建裸鼠分为3组,分别用LyDCs+Tα1、LyDCs和生理盐水皮下免疫注射治疗;治疗后7 d,体外观察各组裸鼠脾脏淋巴细胞的肿瘤杀伤作用及IFN-γ、IL-4分泌水平,实验结束时,观察LyDCs联合Tα1对荷瘤裸鼠的体内抑瘤作用。结果LyDCs的表型HLA-DR、CD80、CD86、CD83较刺激前明显上调;免疫重建裸鼠均检测到人源性CD4^+、CD8^+T细胞;LyDCs+Tα1组裸鼠脾脏T淋巴细胞的肿瘤杀伤作用与LyDCs组比较差异有统计学意义(P<0.01),LyDCs+ Tα1组T细胞的IFN-γ分泌水平与LyDCs组比较差异有统计学意义(P<0.01);接种HT-29细胞58 d后,LyDCs+Tα1组、LyDCs组抑瘤率分别为60.41%、37.20%,两组之间抑瘤效应比较差异有统计学意义(P<0.01),两组的抑瘤效应与对照组比较分别(P<0.01)。结论Tα1能增强LyDCs诱导的CD4^+Th1细胞反应和CTLs杀伤效应,对DCs疫苗的抗癌免疫治疗功效具有明显的放大作用。Objective To investigate the therapeutic effects of thymosin alphal （Tαl） combined with tumor lysate-pulsed dendritic cells （LyDCs） on humanized BALB/C nude mice bearing HT-29 colon carcinoma. Methods LyDCs were routinely prepared from immature DCs loaded with HT-29 tumor cell lysate. Phenotypes of LyDCs pre- or post-stimulated with Tα1 were analyzed by flow cytometry （ FCM ）. Humanized nude mice model bearing colon carcinoma was established by caudal intravenous injection of human peripheral blood T-lymphocytes ,and 2 days later,the levels of CD4^＋ and CD8^＋ T cell subpopulations of human T-lymphocytes in peripheral blood were evaluated by FCM. The tumor models were randomly divided into 3 groups, and immunized by subcutaneous injection of LyDCs plus Tαl, LyDCs or sodium alone, respectively. At day 7 after the last immunization, cytokine release assay and cytotoxicity assay were performed by using spleen T-lymphocytes of the mice. At day 58 after inoculation of tumor cells, the in vivo immunotherapeutic effect on mice was observed. Results The expression of HLA-DR, CD80, CD86 and CD83 in LyDCs was significantly up-regulated after the stimulation with Tα1. Human T-lymphocyte subpopulations, CD4^＋ and CD8^＋ T cells were detected from peripheral blood of humanized nude mice. IFN-γ release and cytotoxicity of CTLs from spleen T-lymphocytes of the mice were both significantly increased （P 〈 0.01 ,respectively） in LyDCs ＋ Tαl group in comparison with those in LyDCs group. The inhibition rate of tumor growth was significantly higher in LyDCs ＋ Tαl group as compared with that in LyDCs group （60.41% vs 37.20% ,P 〈0.01 ）. Conclusion Tcd could strongly enhance CD4 ^＋ Thl immune response and the cytotoxicity effects of antigen specific CTLs induced by LyDCs, thus it may play a crucial role as an ideal adjuvant in DC-based immunotherapy.

关 键 词：结肠癌 树突状细胞 胸腺肽 免疫治疗 

分 类 号：R392[医药卫生—免疫学] R735.35[医药卫生—基础医学]