hTERT多表位肽致敏mDC诱导CTL对HLA-A24^+肿瘤细胞的特异性杀伤作用  

CTLs induced by hTERT-related multiepitope peptide-sensitived mDCs specifically kill HLA-A24^+ tumor cells

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

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆400010 [2]重庆医科大学附属第一医院老年科,重庆400016 [3]解放军324医院消化科,重庆400020

出　　处：《细胞与分子免疫学杂志》2011年第6期626-630,633,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市科委科技攻关资助项目(CSTC2009AC5017)

摘　　要：目的:研究人工合成人端粒酶逆转录酶(hTERT)多表位混合肽经髓样树突状细胞(mDC)提呈后,诱导特异性细胞毒性T淋巴细胞(CTL)对HLA-A24+肿瘤细胞的免疫杀伤效应。方法:人工合成四分支的树状串联hTERT表位肽(MAPs)及其各单表位多肽。取HLA-A24+健康志愿者的外周血,用免疫磁珠分选并培养mDC。用尼龙毛柱纯化T淋巴细胞并培养。以各表位肽致敏mDC后,诱导特异性CTL增殖,并以表达hTERT且以HLA-A24+肿瘤细胞株SMMC-7721及HLA-A24-肿瘤细胞株SKOV3为靶细胞行杀伤实验。用ELISA法检测不同时间点培养基上清液中IL-12、TNF-α的分泌量;用流式细胞术检测CTL对肿瘤细胞的杀伤效应。结果:以源于hTERT的T淋巴细胞表位I540(ILAKFLHWL)、V461(VYGFVRACL)及L766(LTDLQPYMRQFVAHL)合成4分支MAPs多肽及各单表位混合多肽。以人工合成的hTERT多表位混合肽致敏mDC后,能刺激CTL增殖,并可诱导对HLA-A24+肿瘤细胞株SMMC-7721的特异性杀伤作用,且MAPs多肽较单表位混合多肽的致敏效果更具显著性(P<0.05)。结论:以人工合成的hTERT多表位混合肽致敏mDC能激活同源淋巴细胞(CTL),可特异性杀伤HLA-A24+的肿瘤细胞,在肿瘤免疫治疗中具有重要的意义。AIM： To study the antigen specific anti-tumor effect of cytotoxic T lymphocytes（CTLs）,which was induced by human telomerase reverse transcriptase（hTERT）-related multiple epitope peptides impulsed myeloid dendritic cells（mDCs）,against human leukocyte antigen（HLA）-A24＋ tumor cells.METHODS： Four branches of multiple antigen peptides（MAPs） of hTERT epitopes and three separate peptides were solid-phase artificially synthesized,phlebotomize peripheral blood from HLA-A24＋ healthy volunteers,sorted the blood through MACS MicroBeads and cultured mDCs,Nylon fiber column purified T lymphocytes,mDCs impulsed with each type of peptides were co-cultured with T lymphocytes to induce CTLs specifically killing effect,and the resultant CTLs were used as effector cells,SMMC-7721 with hTERT and HLA-A24 positive and SKOV3 which are hTERT-positive but HLA-A24-negative tumor cells were used as target cells.The level of human IL-12,TNF-α in the culture supernatant was determined by ELISA.Flow cytometry assay was used to assess the killing ability of CTLs against tumor cells.RESULTS： MAPs of hTERT epitopes including I540（ILAKFLHWL）,V461（VYGFVRACL）,L766（LTDLQPYMRQFVAHL） and three separate peptides could impulse mDCs and then induce CTLs to specifically kill SMMC-7721,CTLs induced by MAPs had stronger cytotoxic effect compared with three separate peptides mixed（P〈0.05）.CONCLUSION： mDCs-impulsed with hTERT-associated MAPs can induce production and proliferation of allogenic CTLs,which show antigen specific anti-tumor effect against HLA-A24＋ tumor cells.This result has significantly meaning in tumor immunotherapy.

关 键 词：人端粒酶逆转录酶 多抗原表位肽 髓样树突状细胞 HLA-A24+肿瘤细胞 免疫治疗 

分 类 号：R73-36[医药卫生—肿瘤]