SARS患者康复期淋巴细胞功能状态及T细胞受体Vβ24个亚家族表达的格局(英文)  

Lymphocyte immunological function and 24 T cell receptor V beta subfamilies expression in convalescent patients with severe acute respiratory syndrome

作　　者：曾星 蔡萃 黄羽 欧爱华 张娴 

机构地区：[1]广州中医药大学第二附属医院中心实验室,广东省广州市510120 [2]广州中医药大学第二附属医院临床流行病学研究室,广东省广州市510120

出　　处：《中国组织工程研究与临床康复》2007年第43期8796-8800,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省中医药管理局基金资助(203014)~~

摘　　要：背景:严重急性呼吸道综合征(severe acuterespi ratory syndrome,SARS)是由SARS相关冠状病毒感染引起的急性传染性疾病。但目前其康复期患者细胞免疫状态仍有待进一步研究。目的:观察SARS患者康复期淋巴细胞功能状态和T细胞受体(TCR)Vβ24个亚家族表达的格局。设计:前后对照观察。单位:广东省中医院中心实验室。对象:选择2003-01/04在广州中医药大学第二附属医院收治的76例治愈的SARS患者,全部病例均符合"非典型肺炎的临床诊断标准"、"非典型肺炎重症病例诊断标准、出院标准"及《中医临床诊疗术语证候部分》的诊断标准,男30例,女46例,平均(32±11)岁。选择10名同期健康到院体检者,女5名,男5名,平均(32±7)岁,所有受试对象对检测项目知情同意。方法:①TCRVβ24个亚家族的表达检测:取受试对象全血2mL,用EDTA-K2抗凝管抗凝。在流式细胞术检测专用管中,分别加入各种荧光素标记的mAb:抗CD3、抗CD4、抗CD8、抗CD25、抗CD28、抗HLA-DR、PC5-抗CD3mAb、TCRVβ1(PE+FITC)、Vβ2(PE+FITC)、Vβ3(FITC)、Vβ4(PE+FITC)、Vβ5.1(PE+FITC)、Vβ5.2(PE)、Vβ5.3(PE)、Vβ7.1(PE+FITC)、Vβ7.2(FITC)、Vβ8(FITC)、Vβ9(PE)、Vβ11(PE)、Vβ12(FITC)、Vβ13.1(PE)、Vβ13.2(PE)、Vβ13.6(PE+FITC)、Vβ14(FITC)、Vβ16(FITC)、Vβ17(PE+FITC)、Vβ18(PE)、Vβ20(FITC)、Vβ21.3(FITC)、Vβ22(PE+FITC)和Vβ23(PE)10μL,然后加入抗凝血50μL。混匀后,于室温避光孵育15min,再加入溶血素溶血、洗涤。最后将沉淀溶在300μLPBS中,用CoulterESP流式细胞仪进行检测。②T细胞亚群,T、B细胞的活化状态及Ts细胞和Tc细胞百分率的检测:每次收集5000个细胞,用仪器所带软件分别计算T细胞亚群(CD3、CD4和CD8)、活化的T、B细胞(CD3+/CD25+、CD3+/HLA-DR+、和CD3-/HLA-DR+)、Ts细胞和Tc细胞(CD8+/CD28+、CD8+/CD28-)的百分率。主要观察指标:①T细胞亚群(CD3、CD4和CD8)的表达。②活化的T、B细胞(CD3+/CD25+、CD3+/HLA-DR+、和CD3-BACKGROUND： Severe acute respiratory syndrome （SARS） is caused by a genetically novel coronavirus that is caused by acute infectious disease. It is not yet clear for the immunology function of SARS patients in their convalescent stage. OBJECTIVE： To study the effects on T lymphocyte, and the titer profiling of 24 T cell receptor （TCR） V β subfamilies expressions in SARS convalescent patients. DESIGN ： A self-control observation SETTING ： Central Laboratory, Guangdong Provincial Hospital of Traditional Chinese Medicine PARTICIPANTS： Seventy-six cured SARS patients who received treatment in the Second Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine between January and April 2003. All the patients corresponded to ＂clinical diagnostic criteria of atypical pneumonia＂, ＂ diagnostic criteria of severe atypical pneumonia and discharge criteria＂ and ＂clinical diagnostic criteria and discharge criteria of severe acute respiratory syndrome＂. The involved patients, 30 male and 46 female, averaged （32±11）years old. Another 10 subjects who simultaneously received health examination in the same hospital, 5 male and 5 female, aged （32±7）years, were involved in the study. Informed consents of detected items were obtained from all the subjects. METHODS; （1） Detecting the expression of 24 T cell receptor （TCR） V β subfamilies in SARS convalescent patients： Peripheral blood （2 mL） was collected from the healthy convalescent subjects, and EDTA-K2 was used as anticoagulant. In the flow cytometry detection tubes, 10 μL various fluorescein-labeled mAb, such as anti-CD3, anti-CD4, anti-CD8, anti-CD25, anti-CD28, anti-HLA-DR, anti-CD3mAb conjugated with PC5, TCR Vβ1 （PE±FITC）, Vβ2（PE±FITC）, Vβ3 （FITC）, Vβ4（PE±FITC）, Vβ5.1 （PE±FITC）, Vβ5,2（PE）, Vβ5.3（PE）, Vβ7.1 （PE±FITC）, Vβ7.2（FITC）, Vβ8（FITC）, Vβ9 （PE）, Vβ11 （PE）, Vβ12（FITC）, Vβ13.1 （PE）, Vβ13.2（PE）, Vβ13.6（PE±FITC）, V�

关 键 词：SARS 康复期 T细胞亚群 TCRVβ亚家族 

分 类 号：R373.1[医药卫生—病原生物学]