MUC1模拟表位肽对表达人MUC1 T739小鼠膀胱癌细胞的细胞毒作用的实验研究  

Cytotoxicity of MUC1-positive T739 mouse bladder cancer cells induced with MUC1 mimic peptide epitopes

作　　者：崔志刚[1] 刘刚[1] 裘辉[1] 潘进勇[1] 路浩军[1] 张立新[1] 

机构地区：[1]解放军第306医院泌尿外科,北京100101

出　　处：《中华临床医师杂志（电子版）》2012年第2期75-78,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家"863"计划资助项目(2002AA214111)

摘　　要：目的观察MUC1模拟表位肽对表达人MUC1T739小鼠膀胱癌细胞的细胞毒作用。方法培养、诱导T739小鼠骨髓成熟树突状细胞,流式细胞仪鉴定纯度,用负载模拟表位肽成熟树突状细胞免疫T739小鼠,酶联免疫斑点(ELISPOT)检测活化的模拟表位特异性T细胞,用非放射性乳酸脱氢酶法检测不同效:靶比例时活化的模拟表位特异性T细胞体外细胞毒效应。结果培养出成熟树突状细胞,纯度为78.8%,肽免疫的T739小鼠特异性分泌IFN-γ的T细胞频数是106.5±12.8,而PBS和DC+PBS组频数均少于10,数量明显少于肽+DC组。当效靶比为100:1时肽免疫的T739小鼠产生的活化T淋巴细胞具有显著细胞毒功能,与PBS和DC+PBS免疫小鼠相比,差异有统计学意义(P<0.01)。结论负载MUC1模拟表位肽的树突状细胞免疫小鼠,可诱导产生特异性细胞毒性T细胞。Objective To investigate the functional immunological responses and effectiveness in treatment of MUCl-positive T739 mouse bladder cancer induced with MUC1 mimic peptide epitopes. Methods Mouse dendritic cells（DCs） were derived from T739 bone marrow and analyzed on a flow cytometer. Mice were immunized peptide-loaded DCs. Mimotope-specific immune responses were detected by ELISPOT technology. Mimotope-specific CTL responses with different E/T ratio in vitro were also analyzed by non-radiative LDH cytotoxicity analysis. Results Mature DC were harvested and FCM showed thai 78. 8% cells expressed a marker of mature DC. The results of ELISPOT showed that the frequencies of functional IFN-γ secreting lymphocytes in immunized mice with loaded peptide DCs were 106.5±12. 8 and the frequencies in immunized mice with unloaded peptide DCs or PBS were not more than 10, which were significantly lower than experimental group. Non-radiative LDH cytotoxicity analysis manifested that the specific active T lymphocytes of mice immunized with peptide-loaded DCs could significantly kill BST739-MUC1 ceils when the E/T ratio was 100：1 ,which were significantly different from that of mice immunized with unloaded peptide DCs or PBS（P 〈 0. 01 ）. Conclusions Mimotope-specific CTL of mice can be induced with immunized mimic epitope peptide-pulsed DC in vitro.

关 键 词：膀胱肿瘤 CA-15-3抗原 T淋巴细胞 细胞毒性 模拟抗原表位 

分 类 号：R737.14[医药卫生—肿瘤]