HCoV-OC43逃避人树突状细胞免疫清除机制的初步研究  

Preliminary mechanism study of HCoV-OC43 escape from human dendritic cell immune elimination

作　　者：杨权[1] 庹玖玲 黄旭斌[4] 罗洪娇 周凯[2,3] 张甜[2,3] 曹开源[2,3] 徐霖[2,3] 

机构地区：[1]广州医科大学基础学院病原生物学与免疫学教研室,广州511436 [2]中山大学热带病防治研究教育部重点实验室,广州510080 [3]中山大学香港大学粤港传染病监测联合实验室,广州510080 [4]中山大学附属第一医院MICU室,广州510080

出　　处：《中国免疫学杂志》2017年第4期488-493,共6页Chinese Journal of Immunology

基　　金：国家传染病防治科技重大专项(No.2012ZX10004-213);国家自然科学基金项目(No.81000230);广东省自然科学基金青年项目(No.A030310282)资助

摘　　要：目的:了解人冠状病毒OC43(Human coronavirus OC43,HCoV-OC43)逃避人树突状细胞(Dendritic cell,DC)免疫监视作用的初步机制。方法:利用HCoV-OC43阳性病人的标本感染BSC-1细胞分离OC43病毒,相差显微镜观察细胞病变(Cytopathic effect,CPE),实时荧光定量聚合酶链反应(Real-time PCR)进行鉴定;利用人细胞因子GM-CSF和IL-4联合体外诱导DC分化,于诱导7 d后用HCoV-OC43病毒感染DC。采用透射电镜观察DC感染后的形态,Real-time PCR检测DC功能相关细胞因子的表达水平;流式细胞术检测DC比例及其功能相关共刺激分子的表达。结果:成功建立HCoV-OC43体外感染DC的体系。HCoV-OC43能感染DC并刺激其产生免疫应答,但培养上清中不能检测到病毒核酸;HCoV-OC43感染会导致DC细胞表达IFN-α、IFN-β、CCL3和CCL5的量显著下调,但其共刺激分子HLA-DR、CD1c和CD86的表达不受抑制。结论:HCoVOC43可感染人DC细胞并刺激其产生免疫应答,但不能产生活的子代病毒;HCoV-OC43可通过抑制宿主DC细胞IFN-α等相关炎症因子和趋化因子的分泌,来实现免疫逃逸。Objective： To study the possible immune escape mechanisms of HCoV-OC43 from human dendritic cells （DC）. Methods ：HCoV-OCA3 was isolated from clinical specimen using BSC-1 cells and identified by Real-time PCR, and the cytopathic effect was observed by phase contrast microscope. DCs were induced in vivo using hu-GM-CSF and IL-4 eytokines, and after 7 days of differ- entiation,DCs were infected by HCoV-OCA3. The morphology of HCoV-OCA3 infected DC was observed by transmission electron microscope,and the eytokines related to DC functions were detected by Real-time PCR after infection. DC proportion and function related co-stimulatory molecules were analyzed by flow eytometry. Results： In vitro HCoV-OC43 infected human DC model was successfully built. HCoV-OC43 can infect DC and generate immune response of DC in vitro, but no virus nucleonic acid could be detected in culture supernatant. The DC expression of IFN-a, IFN-β, CCL3 and CCL5 were significant decreased when infected with HCoV-OC43,but the expression of costimulatory molecules including HLA-DR, CDlc and CD86 were not affected by HCoV-OC43 infection. Conclusion：Human DC could be infected by HCoV-OC43 and generate immune response, but could not produce progeny virus. HCoV-OC43 may escape from immune response by suppressing the expression of IFN-a and other inflammatory cytokines and chemokines in DC.

关 键 词：人冠状病毒OC43 人树突状细胞 细胞因子 免疫逃逸 

分 类 号：R373.1[医药卫生—病原生物学]