人工合成树状串联hTERT表位肽对mDC表型和功能的影响  

Effect of Synthesized hTERT Epitopes on the Phenotype and Function of Myeloid Dendritic Cells

作　　者：钮柏琳[1] 慎华平[1] 杜慧敏[2] 杨仕明[3] 邹利全[4] 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,重庆市400010 [2]重庆医科大学附属第一医院老年科,重庆市400016 [3]第三军医大学附属西南医院全军消化研究所,重庆市400038 [4]解放军第324医院消化科,重庆市400020

出　　处：《医学分子生物学杂志》2011年第2期143-148,共6页Journal of Medical Molecular Biology

基　　金：重庆市科委科技攻关项目（No.CSTC2009AC5017）

摘　　要：目的 通过对树状串联hTERT表位肽（MAP）与无肽刺激组髓样树突状细胞（mDC）的相关检测,研究MAP肽对mDC的表型和功能的影响.方法 人工固相合成四分支的MAP肽,免疫磁珠分选mDC,流式细胞技术检测相关表面分子.ElISA分别检测两组mDC培养基的IL-12p70含量,并作统计学分析.结果 人工合成MAP肽纯度高（95.26 %）,免疫磁珠分选mDC纯度为72.59 %,相比于无肽刺激组mDC,MAP肽刺激组成熟（培养第9天）时MHCⅡ类分子为83.90 %、MHCⅠ类分子为91.08 %,CD86：77.03 %,CD83：92.16 %;IL-12p70的含量也大于无肽刺激组,且有统计学差异.结论 人工合成hTERT的MAP肽能足够强的激活mDC,刺激其成熟和功能的表达,可作为mDC抗肿瘤疫苗的刺激肽结构.Objective To explore the role of tandem multiple antigenic peptide （MAP） on myeloid dengritic ceils （mDC） , by comparing the phenotype and function of mDC stimulated by MAP of human telomerase reverse transeriptase （hTERT） epitopes to the control group cells. Methods Four branches of MAP peptides were solid-phase artificially synthesized, mDC cells were isolated through magnetic activated cell sorting （MACS） . The surface molecules of mDC were detected by flow cytometry. The level of IL-12 pT0 was detected by ELISA on both groups respectively and statistically analyzed. Results Synthetic MAP peptide was synthesized with high purity （95.26%） . The rate of mDC with magnetic activated cell sorting was 72. 59%. In MAP peptide stimulated group, MHC Ⅱ molecules reached 83.90%, MHCI 91.08 %, CD86 77. 03%, and CD83 92. 16%, while the control group was 12.92%, 64. 88%, 14. 45% and 26. 32%, when matured in 9-day culture. IL-12p70 secretion was higher than that in the control group, with significant difference. Conclusion The synthesized MAP peptides of hTERT can strongly activate mDC, thus can be used as stimulating peptides in anti-tumor vaccine research.

关 键 词：人端粒酶逆转录酶 表位肽 髓样树突状细胞 磁珠分选 无血清培养基 

分 类 号：R341.2[医药卫生—基础医学]