HCMV感染对人THP-1细胞HLA-G及其受体表达的影响

发布时间：2018-04-23 11:06

本文选题：人巨细胞病毒 + 人类白细胞抗原G　；参考：《中国病理生理杂志》2017年02期

【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(P0.01)。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(P0.05)。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。
[Abstract]:Aim: to investigate the effect of human cytomegalovirus (HCMV) infection on the expression of HLA-GG isoforms of human leukocyte antigen (GG) and its receptors in human THP-1 cells and to explore the role of HLA-G in the immune response of HCMV escape host. Methods RT-PCR and Western blot were used to detect the mRNA and protein levels of HLA-G isoforms in THP-1 cells infected with Towne strain. The expression of HLA-G and its surface receptor ILT4 in THP-1 cells were detected by flow cytometry. IL-10 and soluble HLA-GN in the supernatants of THP-1 cells were detected by Elisa. Cell survival rate was also measured. Results there was no obvious apoptosis in the cells infected with HCMV. The mRNA expression of HLA-G1 and HLA-G5 was upregulated 1 d after infection with THP-1 cells. The expression of HLA-G1 and HLA-G5 in THP-1 cells was significantly up-regulated after 1 day of infection. The expression of HLA-G-1 and ILT4 in THP-1 cells increased significantly after 1 day of infection. Compared with the control group, the level of IL-10 in the culture supernatant of P0.01 + THP-1 cells was significantly up-regulated 1 day after infection, and the difference was statistically significant compared with the control group. Conclusion the differential expression of HLA-G isomers was induced by THP-1 cells infected with HLA-G1 and HLA-G5, and the expression of ILT2/ILT4 on the surface was up-regulated. At the same time, HCMV infection can induce THP-1 cells to secrete IL-10. This study provides experimental evidence for further exploring the mechanism of HCMV evading immune response.
【作者单位】：温州医科大学附属第二医院附属育英儿童医院检验科;温州医科大学检验医学院检验医学教育部重点实验室;
【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)
【分类号】：R392

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