树状串联hTERT表位肽负载mDC疫苗体外激发抗瘤免疫应答的效应研究  

Effects of specific lymphocytes activated by mDC loaded with tandem multiple antigenic peptide of hTERT epitopes on anti-tumor cells

作　　者：钮柏琳[1] 慎华平[1] 龚建平[1] 杜慧敏[2] 杨仕明[3] 邹利全[4] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科,400010 [2]重庆医科大学附属第一医院老年科,400016 [3]第三军医大学附属西南医院全军消化研究所,重庆400038 [4]解放军324医院消化科,重庆400020

出　　处：《免疫学杂志》2011年第6期475-481,共7页Immunological Journal

基　　金：重庆市科委科技攻关项目(CSTC2009AC5017)

摘　　要：目的通过树状串联hTERT表位肽的髓样树突状细胞(mDC)递呈,刺激同源淋巴细胞,探索一种更优的肿瘤免疫治疗方法。方法人工固相合成4分支的树状串联hTERT表位肽(MAP)及其各分支单肽,免疫荧光检测hTERT的表达情况,免疫磁珠分选mDC,尼龙毛柱纯化T细胞,ELISA检测mDC的IL-12p70和淋巴细胞的(LC)TNF-α、IFN-γ分泌量,流式细胞技术检测mDC、LC的相关表面分子以及效应性T细胞对HLA-A2型肿瘤A549、MDA-MB-231和SW480的杀伤率,并作统计学分析。结果所有肿瘤细胞都表达hTERT,且胞核大于胞浆;MAP和混合单肽对3种肿瘤细胞都有杀伤效应,且MAP的杀伤效应大于混合单肽,有统计学差异。结论人工合成hTERT的MAP肽通过mDC能足够强的激活同源淋巴细胞,在肿瘤疫苗的研发中有重要意义。Human telomerase reverse transcriptase（hTERT） is,recently discovered,a closely related enzymes of tumors in unlimited proliferation,which highly express in most tumor tissue,but weakly positive or negative in normal tissues.Previous studies have found that inhibition of hTERT activity of cells can inhibit the growth of tumor cells until death,and using hTERT epitope peptide to stimulate dendritic cells（DC） can effectively activate anti-tumor immune response in vitro.But further study results showed that single-epitope synthetic hTERT peptides although can made effective antitumor effects in vitro,there was no significant in vivo.This study taking advantage of synthetic multi-epitope of hTERT（tree series multiple antigen peptide antigen Tandem-MAP,which content three simple peptides I540,V461 and L766） loaded autologous mDC（myeloid dendritic cells） to activate the homologous T-lymph cell,and then detect the killing effect on HLA-A2 tumor cells A549,MDA-MB-231 and SW-480 by flow cytometry.Result showed that tandem-MAP group has the strongest killing effect on the three kinds of tumor cell,compared with the simple mixture of peptides and non peptides stimulated group.Thus,we concludes that the artificial synthesized MAP of in hTERT epitopes presented by mDC can activate the homologous lymphocytes strong enough,and have an effect on a variety of tumor cells,which will be great significance in the tumor vaccine research.

关 键 词：表位肽 髓样树突状细胞 磁珠分选 肿瘤 免疫治疗 

分 类 号：R392[医药卫生—免疫学]