肠道病毒71型衣壳蛋白VP1特异性识别和激活手足口病患儿CD4^+T细胞免疫活化  

Enterovirus 71 capsid protein VP1 specifically recognizes and activates CD4^+ T cell immune activation in children with hand, foot and mouth disease

作　　者：高宁[1] 李梅[1] 李亚萍[1] 王文俊 石娟娟[1] 贾晓黎[1] 党双锁[1] Gao Ning;Li Mei;Li Yaping;Wang Wenjun;Shi Juanjuan;Jia Xiao li;Dang Shuangsuo.(Department of Infectious Diseases,the Second Affiliated Hospital of Medical School of Xi 'an Jiaotong University,Shannxi 710004,China)

机构地区：[1]西安交通大学第二附属医院感染科,西安市710004

出　　处：《中华实验和临床感染病杂志（电子版）》2018年第5期427-433,共7页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学青年科学基金项目(No.81701632);西安交通大学第二附属医院人才培养专项科研青年基金[No.YJ(QN)201520];陕西省重点研发计划项目(No.2017ZDXM-SF-071)

摘　　要：目的探讨肠道病毒病毒71型衣壳蛋白VP1诱导EV71感染患儿外周血T淋巴细胞产生干扰素-γ(IFN-γ)应答特征。方法收集2010年9月至2011年12月西安交通大学医学院第二附属医院和西安市儿童医院住院的EV71感染手足口病患儿共31例,根据临床特征分为3组:急性感染期组(16例)、临床恢复期组(9例)和感染恢复后1个月组(6例);另外,选取10例健康儿童为对照组。通过重叠肽技术合成覆盖EV71衣壳蛋白VP1 36条重叠肽段作为抗原,采用IFN-γ酶联免疫斑点分析法(ELISPOT)和磁珠分选法(MACs)体外检测各组患儿EV71 VP1特异性CD4^+T细胞和CD8+T细胞免疫应答特征。结果急性感染期组有4例(25%)患儿对VP1蛋白存在T细胞免疫反应,临床恢复期组5例(56%)患儿对VP1蛋白存在T细胞免疫反应,其中EV71感染临床恢复期组患者反应强度最强,显著高于急性感染期,差异有统计学意义(Z=-2.042、P=0.041),而感染恢复后1个月组和对照组均未检测到T细胞免疫反应,临床恢复期组患者T细胞反应强度显著高于感染恢复后1个月组(Z=-2.105、P=0.035)和对照组(Z=-2.633、P=0.008),差异均有统计学意义;提示EV71感染患儿VP1蛋白被识别的频率(即例数)随着临床症状及体征消失而消失。VP1蛋白诱导的细胞免疫应答以特异性CD4^+T细胞为主导;筛选确定了8个EV71 VP1蛋白区CD4^+T细胞表位。结论 EV71感染患儿在发病期对VP1蛋白存在特异性T细胞反应,且以特异性CD4^+T细胞为主导;筛选确定EV71感染患儿VP1蛋白的CD4^+T细胞表位有助于阐明EV71感染的细胞免疫学发病机制。Objective To investigate the characteristics of interferon-γ(IFN-γ) response induced by enterovirus 71(EV71) capsid protein VP1 in peripheral blood T lymphocytes of children with EV71 infection. Methods From September 2010 to December 2011, a total of 31 patients with hand, foot and mouth disease(HFMD) infected with EV71 were collected from the Second Affiliated Hospital of Medical College of Xi’an Jiaotong University and Xi’an Children’s Hospital. According to the clinical features, 31 cases were divided into three groups: acute infection group(16 cases), clinical convalescence group(9 cases) and one month after recovery of infection group(6 cases), in addition, 10 healthy children were selected as control group. Total of 36 overlapping peptides of EV71 capsid protein VP1 were synthesized by overlapping peptide technique as antigens. The immunological responses of EV71 VP1 specific CD4^+ T and CD8+ T cells were detected by IFN-γ enzyme-linked immunoblot assay(ELISPOT) and magnetic cell sorting(MACs) in vitro. Results In the acute infection group, 4 cases(25%) had T cell immune response to VP1 protein, 5 cases(56%) in the clinical convalescence group had T cell immune response to VP1 protein. The strength of T-cell responses in clinical convalescence group had the strongest, significantly higher than acute infection group(Z =-2.042, P = 0.041), and one month after recovery of infection group(Z =-2.105, P = 0.035) and control group(Z =-2.633, P = 0.008), with significant differences. The T cell immune response was not detected in the control group and one month after recovery of infection group. The results suggested that the frequency of VP1 protein recognition(the number of cases) disappeared with the disappearance of clinical symptoms and signs in children with EV71 infection. The cellular immune response induced by VP1 protein was dominated by specific CD4^+ T cells. Eight CD4^+ T cell epitopes of EV71 VP1 protein region were identified. Conclusions There was a specific T cell response to VP1 protein

关 键 词：肠道病毒71 T细胞免疫 衣壳蛋白VP1 酶联免疫斑点分析 T细胞表位 

分 类 号：R725.1[医药卫生—儿科]