人疱疹病毒6型特异性CD4^+ T细胞的初步克隆  

Establishment and functional characterization of HHV-6 specific CD4^+ T cell clones

作　　者：王芳[1] 姚堃[2] 冯冬举[2] 周锋[2] 尹全章[2] 

机构地区：[1]南京医科大学第一附属医院(江苏省人民医院)临床检验中心 [2]南京医科大学微生物学与免疫学系,南京210029

出　　处：《微生物与感染》2008年第1期17-20,共4页Journal of Microbes and Infections

基　　金：国家自然科学基金30771961

摘　　要：目的克隆人疱疹病毒6型(HHV-6)特异性CD4+ T细胞,并分析其基本特征。方法采用微孔有限稀释法克隆HHV-6特异性CD4+ T细胞;3H-TdR掺入法细胞增殖试验筛选HHV-6特异性CD4+ T细胞克隆;流式细胞仪(FACS)分析HHV-6特异性CD4+ T细胞克隆的表面标志,酶联免疫吸附试验(ELISA)检测HHV-6特异性CD4+ T细胞克隆的细胞因子分泌水平。结果获得的细胞克隆中有4株以HHV-6感染细胞裂解物特异的方式增殖,并且其增殖水平与HHV-6感染细胞裂解物呈剂量依赖性;细胞克隆表面标志为CD3+CD4+;W-2和W-4细胞克隆的细胞因子分泌以白细胞介素10(IL-10)为主,W-1细胞克隆的细胞因子分泌以IL-10及γ干扰素(IFN-γ)为主,W-3细胞克隆的细胞因子分泌以IFN-γ为主。结论初步建立HHV-6特异性CD4+ T细胞克隆,并分析其表面标志和细胞因子分泌水平,为HHV-6特异性Treg细胞的克隆、筛选及其功能的研究奠定基础。Objective To establish HHV-6-specific CD4^＋ T cell clones and to study the characteristics of HHV-6-spocific CD4^＋ T cells. Methods We obtained CD4^＋ T cell clones by using the liquid microwell limiting dilution technique, and selected HHV-6-spocific CD4^＋ T cell clones by means of ^3H-thymidine uptake. The phenotype of the HHV-6-specific T cell clones was analyzed with FACS. We determined IL-10,IL-4 and IFN-γ secretion from HHV-6-specific CD4^＋ T cell clones by ELISA.Results Of the five HHV-6-spocific CD4^＋ T cell clones, four showed proliferative responses to stimulation with HHV-6-infected JJHAN lysates. Moreover, proliferation responses of HHV-6- specific CD4^＋ T cell clones depended upon the concentration of the HHV-6-infected JJHAN lysates. The phenotypic analysis with FACS demonstrated that HHV-6-specific T cell clones consisted of CD3^＋ CD4^＋ T cells. The CD4^＋ T cell clones W-2 and W-4 had Trl-type profiles while the CD4^＋ T cell clone W-1 had a Th0-type profile and the clone W-3 had a Thl-type profile. Conclusion In the current study,we established HHV-6-specific CD4^＋ T cell clones in order to examine their particular phenotype, the nature of cytokine secretion, and to provide an experimental basis for selecting HHV-6-specific CD4^＋ Treg cell clones.

关 键 词：人类疱疹病毒6型 克隆 CD4^+ T细胞 

分 类 号：R392[医药卫生—免疫学]