HPV18 E77-15免疫原性的实验研究  

Immunogenicity of the peptide HPV18 E77-15

作　　者：张乾[1] 徐云升[2] 朱雪洁[1] 李晓林[1] 杨孝军[1] 欧荣英[1] 郑飞云[1] 

机构地区：[1]温州医学院附属第一医院妇产科,325000 [2]温州医学院附属第一医院皮肤科,325000

出　　处：《浙江医学》2012年第4期240-243,共4页Zhejiang Medical Journal

基　　金：国家自然科学基金（30500537,30972800）

摘　　要：目的鉴定抗原肽HPV18E77-15能否诱导HLA—A2＋健康供者的外周血淋巴细胞（PBL）产生抗原肽特异性细胞毒性T细胞（CTL），判断其是否具有免疫原性。方法通过体外细胞培养技术分为两组，实验组：负荷抗原肽为HPV18 E77-15，对照组：负荷的抗原肽为HBVc17-28，采用抗原肽负荷外周血单个核细胞（PBMC）及T2细胞作为抗原呈递细胞，重复刺激HLA—A2阳性健康供者的外周血淋巴细胞（PBLs），1周1次，共3次；分别在第1轮刺激后和第3轮刺激后，采用RPE标记的抗原肽特异性五聚体和FITC标记的CD8＋抗体通过流式细胞仪检测抗原肽所诱导产生的抗原肽特异性CTL的频率。结果在流式细胞仪检测中，实验组第1轮刺激后所选淋巴细胞群中CD8＋五聚体＋双阳性的抗原肽特异性CTL为（1．87±001）％，第3轮刺激后抗原肽特异性CTL增至（15．73±2．47）％；而对照组中未检出抗原肽特异性的CTL细胞增殖。结论抗原肽HPV18E77-15能够诱导产生抗原肽特异性CTL增殖，具有良好的免疫原性，可能成为针对HPVl8感染的治疗性多肽疫苗的候选表位。Objective To identify the immunogenicity of peptide HPV18 E77-15 to induce activation and proliferation of peptide-specific cytotoxic T lymphocytes （CTLs）. Methods Peripheral blood lymphocytes （ PBLs ） from healthy HLA-A2（ ＋ ） donors were loaded with peptide HPV18 E77-15 （test group） or HBVc17-28 （control group ）, PBLs were stimulated three times at weekly intervals with the peptide HPV18 E77-15 loaded autologous PBMC and T2 cells. The frequency of the peptide-specific CTLs in cultured cells were detected after the first and third stimulation with the FITC-CD8＋ and RPE-HLA-A2/HPV18 E77-15 entamer in flow cytometric analysis. Results Flow cytometric analysis showed that the frequency of CD8＋ Pentamer＋double positive peptide-specific CTLs of test group increased from 1.87 ± 0.01% after the first stimulation to 15.73 ± 2.47% after the third stimulation, but in control group the frequency of the peptide-specific CTLs did not increased. Conclusion Peptide HPV18 E77-15 can induce the peptide-specific CTLs from PBLs of healthy HLA-A2（＋） donors in vitro, which implies HPV18 E77-15 might be used as the candidate epitope for the preparation of peptide-based vaccine for prevention of HPV 18 infection.

关 键 词：HPV18 抗原肽 细胞毒性T淋巴细胞 五聚体 

分 类 号：R737.33[医药卫生—肿瘤]