US31介导人巨细胞病毒调控系统性红斑狼疮发生的机制研究  

Study on mechanism of US31 mediated human cytomegalovirus in regulation of systemic lupus erythematosus

作　　者：刘春雅[1] 金晶 李玉芳[1] 喻敏[1] 姜毅[1] 徐利鸳[1] LIU Chun-Ya;JIN Jing;LI Yu-Fang;YU Min;JIANG Yi;XU Li-Yuan(Department of Nephrology,People's Hospital of Quzhou,Quzhou 324000,China)

机构地区：[1]衢州市人民医院肾内科,衢州324000 [2]温州医科大学检验医学院、生命科学学院,检验医学教育部重点实验室,浙江省医学遗传学重点实验室,温州325035

出　　处：《中国免疫学杂志》2021年第24期3015-3021,共7页Chinese Journal of Immunology

基　　金：浙江省自然科学基金(LY13H070006);衢州市科技计划项目(2015070)。

摘　　要：目的:研究人巨细胞病毒(HCMV)基因US31调控系统性红斑狼疮(SLE)发生发展的机制。方法:征集临床60例SLE患者和111例健康志愿者,检测SLE患者与健康志愿者血清中抗HCMV-IgG及IgM抗体滴度,检测外周血白细胞中HCMV UL55基因表达,聚类分析SLE患者外周血单个核细胞(PBMC)中HCMV病毒基因表达谱,鉴定在SLE患者中相对特异性高表达的病毒基因。另外,构建过表达US31基因的重组腺病毒(AdUS31),感染THP1单核细胞和THP1源性的巨噬细胞,分析US31基因对单核-巨噬细胞功能的影响。其次,筛选与US31相互作用的蛋白,细胞水平检测NF-κB相关通路蛋白变化,炎症相关基因TNF-α、IL-8、CCL2、ICAM1变化。分离细胞核和细胞质中的蛋白质,检测US31蛋白细胞定位,细胞水平感染过表达US31蛋白的重组腺病毒后抑制MG132蛋白酶表达,检测NF-κB相关通路蛋白变化。结果:SLE患者的抗HCMV-IgG及IgM抗体滴度均明显高于对照组(P<0.05),SLE患者外周血白细胞HCMV检测阳性率明显高于健康志愿者(P<0.001),全转录组测序和mRNA测序显示US31是SLE患者相对特异高表达的病毒基因,进一步研究显示,SLE患者PBMC中US31表达水平明显高于健康对照人群(P<0.001)。炎症相关基因TNF-α、IL-8、CCL2、ICAM1表达量升高(P<0.001)。NF-κB非经典途径的NF-κB2及前体p100可与US31直接相互作用,US31表达可明显促激活状态的NF-κB2(p52)表达。结论:HCMV诱导US31的表达通过与NF-κB通路相互联系直接改变炎症相关基因的表达,进而调控SLE发生。Objective:To explore the mechanism of human cytomegalovirus gene US31 regulating the occurrence and development of systemic lupus erythematosus(SLE).Methods:For clinical 60 cases of patients with SLE and 111 cases of healthy volunteers to detect anti HCMV-IgG and IgM titers in serum,detected HCMV UL55 gene expression in peripheral blood leukocytes,clustering analysis the HCMV virus gene expression profile of peripheral blood mononuclear cell(PBMC)in patients with SLE,and identified relative specificity high expression genes in SLE patients.In addition,recombinant adenovirus that overexpressed US31 protein(AdUS31)was constructed to infect THP1 monocytes and THP1-derived macrophages.Effect of US31 gene on function of mononuclear macrophages was analyzed.Next,screened proteins interacting with US31,and tested changed of NF-κB pathway related proteins,detected changes of inflammation-related genes of TNF-α,IL-8,CCL2 and ICAM1.Proteins in nucleus and cytoplasm were isolated,and the location of US31 protein was detected.Cells transfected recombinant adenovirus that overexpressed US31 protein,MG132 protease inhibitor was added,detected expressions of NF-κB pathway-related proteins.Results:Titers of anti-HCMV-IgG and IgM antibodies in SLE patients were significantly higher than those in control group(P<0.05),positive rate of peripheral blood leukocyte HCMV in SLE patients was significantly higher than that in healthy volunteers(P<0.001),total transcriptome sequencing and mRNA sequencing showed that US31 was a relatively specific and highly expressed virus gene in SLE patients.Further research showed that expression of US31 in PBMC of SLE patients was significantly higher than that in healthy controls(P<0.001).Expressions of inflammation-related genes TNF-α,IL-8,CCL2 and ICAM1 were increased(P<0.001).Moreover,NF-κB2 and its precursor p100 could directly interact with US31.Expression of US31 could significantly promote the expression of NF-κB2(p52)in the activated state.Conclusion:HCMV-induced high expression of US31

关 键 词：人巨细胞病毒 US31 系统性红斑狼疮 

分 类 号：R593.241[医药卫生—内科学]