应用MHC肽四聚体技术检测类风湿性关节炎患者CCP抗原特异性T细胞及初步临床应用  

Application of MHC peptide tetramer to study CCP antigen-specific T-cells in patients with rheumatoid arthritis and its preliminary clinical application

作　　者：房国祥[1] 虞伟 FANG Guoxiang;YU Wei(Department of Clinical Laboratory,Dean Hospital of Changzhou City,Jiangsu,Changzhou 213000,China;Department of Clinical Laboratory,General Hospital of Eastern Theater Command,PLA.,Jiangsu,Nanjing 210000,China)

机构地区：[1]江苏省常州市德安医院检验科,江苏常州213000 [2]东部战区总医院基础医学实验室,江苏南京210000

出　　处：《中国医药科学》2022年第19期38-41,共4页China Medicine And Pharmacy

摘　　要：目的应用MHC肽四聚体技术检测类风湿性关节炎(RA)患者的环瓜氨酸肽(CCP)抗原特异性T淋巴细胞(CCP/AST)。方法选取东部战区总医院2018年4—11月住院及门诊就诊的患者,包括RA组35例,疾病对照组19例,健康对照组30名,分离外周血淋巴细胞,使用流式细胞仪结合肽四聚体技术检测,用于特异性CCP/AST的研究。结果RA组患者的CD4+CD8-tetramer-CCP阳性细胞比率、CD4+CD8-tetramer-CCP阳性细胞比率与CD4+CD8-tetramer-hCLIP阳性细胞比率的差值(CCP/AST)均高于疾病对照组、健康对照组(P<0.05)。RA组中,抗CCP抗体阳性组的CD4+CD8-tetramer-CCP阳性细胞比率、CCP/AST均高于抗CCP抗体阴性组(P<0.05)。应用流式细胞仪结合肽四聚体技术检测的CCP/AST细胞比率用于诊断RA时ROC曲线下面积(AUC^(ROC))为0.823,95%可信区间为0.695~0.954。CCP/AST比率为0.63%时用于RA诊断的敏感度为0.788,特异性为0.882,达到最高的诊断效能。依据CCP/AST的细胞比率是否大于0.63%而将RA患者分为CCP/AST细胞比率非增高与增高两组,两组的肿痛关节数、破坏关节数、红细胞沉降率(ESR)及C反应蛋白(CRP)的数据差异无统计学意义(P>0.05),CCP/AST升高组的类风湿因子(RF)高于CCP/AST非升高组,差异有统计学意义(P<0.05)。结论流式细胞仪结合肽四聚体技术可用于RA患者的CCP/AST研究,且方法敏感、特异、稳定,作为RA的潜在诊断用指标与传统指标联合应用可提高RA的临床诊断水平。Objective MHC peptide tetramer technology was used to study cyclic citrullinated peptide(CCP)antigen-specific T lymphocytes(CCP/AST)in patients with rheumatoid arthritis(RA).Methods From April to November 2018,the inpatients and outpatients of the General Hospital of Eastern Theater Command,PLA.,were selected,including 35 patients in RA group,19 cases in disease control group and 30 cases in healthy control group.Peripheral blood lymphocytes were isolated.MHC peptide tetramer staining and flow cytometry were used for the study of specific CCP/AST.Results The frequency of CD4+CD8 CCP tetramer positive cells,the difference between the frequency of CD4+CD8 CCP tetramer positive cells and the frequency of CD4+CD8 hclip tetramer positive cells(i.e.CCP/AST)in rheumatoid arthritis group were higher than those in disease control group and healthy control group(P<0.05).In rheumatoid arthritis group,the frequency of CD4+CD8 CCP tetramer positive cells and CCP/AST in anti CCP antibody positive group were higher than those in anti CCP antibody negative group(P<0.05).When the frequency of CCP/AST detected by MHC peptide tetramer flow cytometry was used in the diagnosis of RA,the area under the ROC curve(AUC^(ROC))was 0.823 and the 95%confidence interval was 0.695~0.954.When the frequency of CCP/AST was 0.63%,the sensitivity and specificity for RA diagnosis were 0.788 and 0.882,reaching the highest diagnostic efficiency.RA patients were divided into two groups according to whether the cell frequency of CCP/AST was greater than 0.63%.There was no significant difference in the number of swollen and painful joints,the number of damaged joints,ESR and CRP between the two groups(P>0.05).The RF of the CCP/AST elevated group was higher than that of the CCP/AST non elevated group(P<0.05).Conclusion MHC peptide tetramer staining flow cytometry can be used for CCP/AST research in patients with RA,and the method is sensitive,specific and stable,serving as a potential diagnostic index of RA.The combination of potential diagnostic index an

关 键 词：环瓜氨酸肽 类风湿性关节炎 四聚体染色 抗原特异性T淋巴细胞 

分 类 号：R446[医药卫生—诊断学]