系统性红斑狼疮患者外周血单个核细胞中HCMV基因表达谱及其特异性抗体特征  

Expression Profile of HCMV Genes and Specific Antibody Characteristics in the Peripheral-blood Mononuclear Cells of Systemic Lupus Erythematosus Patients

作　　者：施昕妤 谢尚丹 项超丽 李宝青[3] 陈朝生[4] 薛向阳 章慧娣[4] SHI Xinyu;XIE Shangdan;XIANG Chaoli;LI Baoqing;CHEN Chaosheng;XUE Xiangyang;ZHANG Huidi(Institute of Molecular Virology and Immunology,Department of Microbiology and Immunology,School of Medicine,Wenzhou Medical University,Wenzhou 325000,China;Institute of Immunology,Zhejiang University School of Basic Medicine,Hangzhou 310000,China;The Second Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China;First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学基础医学院、分子病毒学与免疫学研究所、微生物与免疫学教研室,温州325000 [2]浙江大学医学院免疫研究所,杭州310000 [3]温州医科大学附属第二医院,温州325000 [4]温州医科大学附属第一医院,温州325000

出　　处：《病毒学报》2023年第1期113-123,共11页Chinese Journal of Virology

基　　金：国家自然科学基金(项目号:82001705),题目:人巨细胞病毒US31基因促进SLE患者巨噬细胞NFKB2活化的机制研究;浙江省医药卫生科技项目(项目号:2019KY453),题目:HCMV US31促进系统性红斑狼疮M1型巨噬细胞极化及分子机制。

摘　　要：人类巨细胞病毒(Human cytomegalovirus,HCMV)感染是系统性红斑狼疮(Systemic lupus erythematosus,SLE)的重要病因,并会加剧疾病进展。然而,SLE患者外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)中HCMV基因的表达谱及其特异性抗体特征尚未完全阐明,并且HCMV蛋白特异性抗体水平与SLE患者临床特征的相关性尚未得到证实。通过Poly(A)建库的mRNA转录组测序(Poly(A)RNA-Seq)检测3例SLE患者和3例健康对照者(Healthy control,HC)的PBMC中的HCMV基因表达谱。然后在10例SLE患者和10例HC的链特异性建库的mRNA转录组测序(strand-specific RNA-seq)结果中验证检测到的HCMV基因。除此之外,通过免疫信息学分析筛选HCMV基因的B细胞表位。ELISA用于检测120例SLE患者和75例HC血清中的HCMV特异性抗体水平,并将其与患者的临床特征相关联。本研究在SLE患者和HC的PBMC中检测到8个HCMV基因。免疫信息学分析筛选出7个HCMV基因的优势B细胞表位。与HC相比,SLE患者血清中UL32和UL82特异性抗体显著降低。UL32特异性抗体可能与SLE患者血细胞功能异常有关。UL82特异性抗体可能参与SLE患者免疫系统功能异常和肾脏功能障碍。受试者工作特征曲线(Receiver operating characteristic,ROC)分析表明,其中UL32特异性抗体可以有效区分SLE患者和HC。Human cytomegalovirus(HCMV) infection is an important cause of systemic lupus erythematosus(SLE) and can exacerbate disease progression. However, the profile of HCMV genes expressed in the peripheral-blood mononuclear cells(PBMCs) of SLE patients and their specific antibody characteristics have not been elucidated fully. Also, the correlation between HCMV gene-specific antibody levels and the clinical characteristics of SLE patients has not been demonstrated. The expression profile of HCMV genes in the PBMCs of three SLE patients and three healthy controls(HCs) was measured by mRNA transcriptome sequencing(Poly(A)RNA-Seq) built by Poly(A). Then, the detected HCMV genes were verified in the results of the strand-specific RNA-sequencing of 10 SLE patients and mRNA transcriptome sequencing(strandspecific RNA-sequencing) of 10 HCs. In addition, the B-cell epitope of the HCMV gene was screened by immunoinformatics analysis. Enzyme-linked immunosorbent assays were used to measure HCMV-specific antibody levels in the sera of 120 SLE patients and 75 HCs, and we correlated them with the clinical characteristics of patients. We detected eight HCMV genes in the PBMCs of SLE patients and HCs.Immunoinformatics analysis screened-out seven dominant B-cell epitopes of HCMV genes. Compared with HCs, levels of UL32-and UL82-specific antibodies in the sera of SLE patients were reduced significantly. UL32-specific antibodies may be related to abnormal function of blood cells in SLE patients. UL82-specific antibodies may be involved in abnormal function of the immune system and renal dysfunction in SLE patients. Analyses of receiver operating characteristic curves showed that UL32-specific antibodies can distinguish SLE patients from HCs.

关 键 词：人巨细胞病毒 系统性红斑狼疮 外周血单个核细胞 B细胞表位 

分 类 号：R392[医药卫生—免疫学] R373.9[医药卫生—基础医学]