HCMV感染对人THP-1细胞HLA-G及其受体表达的影响  

HCMV-infected human THP-1 cells induce expression of HLA-G and its receptors

作　　者：赖媚媚[1,2] 周秋菊[1,2] 楼韵燕 郭彬瀚 王慧燕[1,2] 郑晓群[1,2] 

机构地区：[1]温州医科大学附属第二医院、附属育英儿童医院检验科,浙江温州325027 [2]温州医科大学检验医学院,检验医学教育部重点实验室,浙江温州325035

出　　处：《中国病理生理杂志》2017年第2期329-335,共7页Chinese Journal of Pathophysiology

基　　金：浙江省医药卫生课题项目(No.2017KY474);温州市科技计划项目(No.Y20150304)

摘　　要：目的:研究人巨细胞病毒(HCMV)感染对人THP-1细胞人类白细胞抗原G(HLA-G)异构体及其受体表达的影响探讨HLA-G在HCMV逃逸宿主免疫应答中的作用。方法:HCMV Towne株感染THP-1细胞后,采用RT-PCR和Western blot检测HLA-G异构体mRNA和蛋白水平,流式细胞术检测THP-1细胞HLA-G及其表面受体ILT2、ILT4的表达,ELISA检测细胞培养上清中IL-10及可溶性HLA-G(sHLA-G)水平,同时检测细胞存活率。结果:HCMV感染后细胞未出现明显凋亡,细胞存活率高。HCMV感染THP-1细胞1 d后HLA-G1、-G3、-G4和-G5的mRNA表达明显上调,HLA-G1和HLA-G5的蛋白表达明显上调。THP-1细胞HLA-G、ILT2和ILT4的表达在感染1 d后明显上调。sHLA-G水平在感染1 d后显著升高,与对照组比较差异有统计学意义(P<0.01)。THP-1细胞培养上清液IL-10水平在感染1 d后明显上调,与对照组比较,差异有统计学意义(P<0.05)。结论:HCMV感染THP-1细胞能诱导HLA-G异构体的差异表达,以HLA-G1和HLA-G5为主,且上调其表面受体ILT2/ILT4的表达。同时,HCMV感染能诱导THP-1细胞分泌IL-10。该研究为进一步探讨HCMV逃避机体免疫应答的机制提供实验依据。AIM： To investigate the differential expression of human leukocyte antigen-G （HLA-G） isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus （HCMV） infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism. METHODS： THP-1 cells were infected with HCMV Towne strain. The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively. The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry. The levels of soluble HLA-G （sHLA-G） and IL-10 were measured by ELISA. RESULTS： After infection of the THP-1 cells with HCMV, no obvious apoptosis in the cells was observed, and the viability of the cells was high. A significant up-regulation of HLA-G1, -G3, -G4 and -G5 at mRNA expression level 1 d after infection was found, while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected. The expression of HLA-G/ILT2/ILT4 was evi-dently up-regulated 1 d after infection. The level of sHLA-G was significantly increased 1 d after infection as compared with control group （ P 〈 0. 0 1 ） . The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group. CONCLUSION： The differential expression of HLA-G isoforms and secretion of the receptors ILT2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection. This study provides experimental evidence for evaluating the immune mechanism of HCMV infection.

关 键 词：人巨细胞病毒 人类白细胞抗原G 抑制性受体 感染 

分 类 号：R392.32[医药卫生—免疫学]