负载MtHSP70-PIGR的树突状细胞疫苗对子宫内膜腺癌的抑制作用  

Inhibition of endometrial adenocarcinoma by DC vaccine loaded with MtHSP70-PIGR

作　　者：周明书[1] 刘军[1] 白玉青[1] 孙芳俐 张震宇[2] 李丽[1] ZHOU Ming-shu;LIU Jun;BAI Yu-qing;SUN Fang-li;ZHANG Zhen-yu;LI Li(Department of Obstetrics and Gynecology,Beijing Ditan Hospital Affiliated to Capital Medical University,Beijing,100015,China;Department of Obstetrics and Gynecology,Beijing Chaoyang Hospital Affiliated to Capital Medical University,Beijing,100027,China)

机构地区：[1]首都医科大学附属北京地坛医院妇产科,北京100015 [2]首都医科大学附属北京朝阳医院妇产科,北京100027

出　　处：《中华肿瘤防治杂志》2021年第6期420-427,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家科技部对外合作项目(2012DFA30490)。

摘　　要：目的本研究旨在合成结核分枝杆菌热休克蛋白70-多聚免疫球蛋白受体融合蛋白(MtHSP70-PIGR),以此融合蛋白负载树突状细胞,激活T淋巴细胞,探讨激活的细胞毒性T淋巴细胞(CTL)对子宫内膜腺癌的抑制效应。方法从足月妊娠剖宫产妇脐带血中提取树突状细胞(DC)和T淋巴细胞。以PIGR为靶抗原,将其与MtHSP70羧基端连接合成融合蛋白MtHSP70-PIGR,分别以MtHSP70、PIGR、MtHSP70-PIGR、MtHSP70+PIGR、磷酸缓冲盐溶液(PBS)负载DC,并依次标记为A、B、C、D和E组,E组为对照组。采用流式细胞仪检测各组DCs表面CD83、CD80、CD11c和HLA-DR分子的表达。并应用各组DC激活T淋巴细胞,获得CTL。建立子宫内膜腺癌雌性NOG小鼠模型,应用各组DC疫苗激活的CTL,于第13和24天对小鼠进行2次免疫注射,观察各组肿瘤生长曲线。采用单因素方差分析、SNK检验和Games-Howell法进行统计分析。结果合成的MtHSP70-PIGR融合蛋白纯度>90%。流式细胞检测结果提示,与对照组(E组)相比较,C组DCs表面CD83显著增高(P=0.012),CD80、HLA-DR、CD11c均增高,但差异无统计学意义;C组DCs表面的HLA-DR高于A、B、D组(P=0.007,P<0.001,P<0.001),C组CD83高于A组和D组(P=0.002,P=0.023),差异有统计学意义,CD80、CD11c各组表达差异无统计学意义。小鼠免疫治疗结果显示,从第33天开始,C组肿瘤生长受到抑制,呈现出低于其他各组的趋势。第33天时,C和E组的平均肿瘤体积分别为1208.29303和1448.55025 mm3,C组低于E组(P=0.043),抑瘤率为29.012%;第40天时,C和E组平均肿瘤体积分别为1451.02357和2133.67271 mm3,C组低于E组(P=0.013),抑瘤率为31.994%。结论MtHSP70-PIGR融合蛋白能够促进DC表型成熟,负载MtHSP70-PIGR融合蛋白的DC活化的CTL具有更强的抑制子宫内膜腺癌生长的功效。Objective Mycobacterium tuberculosis heat shock protein 70-Polymeric immunoglobulin receptor(MtHSP70-PIGR)was synthesized to load dendritic cells and activate T lymphocytes,to study the inhibitory effect of activated cytotoxic T lymphocytes(CTL)on endometrial adenocarcinoma.Methods Dendritic cell(DC)and T lymphocytes were extracted from umbilical cord blood of pregnant women undergoing term cesarean section.The fusion protein MtHSP70-PIGR was synthesized by binding polymeric immunoglobulin receptor(PIGR)with the carboxyl terminal of Mycobacterium tuberculosis heat shock protein 70(MtHSP70).DC was respectively loaded on MtHSP70,PIGR,MtHSP70-PIGR,MtHSP70+PIGR and phosphate buffer saline(PBS),marked as Group A,Group B,group C,group D,and group E in turn.Group E was referred as the control group.The expressions of CD83,CD80,CD11 c and HLA-DR molecules on the surface of DCs in each group were detected by flow cytometry.CTL was obtained by activating T lymphocytes with DC in each group.Mice model of endometrial adenocarcinoma with Ishikawa cell was established.Mice were immunized by CTL activated DC vaccine at day13 and 24,and the tumor growth curve for each group was recorded.One-way ANOVA,SNK test and Games-Howell method were conducted for statistical analysis.Results The purity of the synthesized MtHSP70-PIGR fusion protein was>90%.The results of flow cytometry showed that compared with the control group(group E),the expression of CD83 on DC surface in group C was significantly higher(P=0.012),CD80,HLA-DR and CD11 cwere all increased,but the differences were not statistically significant.The expression of HLA-DR on DCs surface in group C was significantly higher than that in group A,B and D(P=0.007,P<0.001,P<0.001),CD83 in group C was higher than that in group A and D(P=0.002,P=0.023),and there was no significant difference for the expression of CD80 and CD11 c.From day 33,tumor growth was inhibited in group C,showing a lower trend than other groups.At day 33,the mean tumor volumes of group C and group E were 1208.2

关 键 词：树突状细胞疫苗 多聚免疫球蛋白受体 结核分枝杆菌热休克蛋白70 子宫内膜腺癌 

分 类 号：R737.33[医药卫生—肿瘤]