猪血凝性脑脊髓炎病毒抑制神经细胞干扰素的产生  

Porcine hemagglutinating encephalomyelitis virus inhibits the production of IFN-β in nerve cell 2

作　　者：岳慧青[1] 杨雅雯[1] 姜丹[2] 张竞[1] 石俊超 潘伟[1] 贺文琦[1] 高丰[1] 宋德光[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]四平梨树县动物疫病预防控制中心,吉林四平136500

出　　处：《中国兽医学报》2017年第4期601-606,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31472194;31602018);吉林省青年科学基金资助项目(20160520033JH)

摘　　要：猪血凝性脑脊髓炎病毒(PHEV)属于β属冠状病毒成员,其具有典型的嗜神经性。多种冠状病毒可逃避宿主先天性免疫的识别而表现出明显的致病性,但宿主的先天性免疫在PHEV感染过程中发挥的作用还不是很清楚。为了明确神经细胞在PHEV感染过程中细胞自身的免疫应答状况,本研究对参与先天性免疫的重要炎性细胞因子和相关的通路变化情况进行了检测分析。用酶联免疫吸附试验(ELISA)方法对PHEV感染小鼠脑神经瘤母细胞(N2a)不同时间点的样品进行检测,结果证明没有IFN-β的表达,说明PHEV不能引起神经细胞产生Ⅰ型IFN。收集感染N2a细胞24h的上清,ELISA方法检测TNF-α、IL-1β、IL-6和IFN-γ的分泌情况,仅有IL-6呈现高水平表达,说明PHEV感染N2a细胞诱发了IL-6的先天性免疫反应。免疫荧光方法检测PHEV感染N2a细胞24h后重要核转录因子IRF-3和NF-κB的核定位情况,发现NF-κB移位到了胞核,IRF-3仍然存在于胞浆,说明PHEV仅激活了NF-κB信号通路。以上试验结果表明,PHEV进入中枢神经系统时可逃避神经细胞以IFN-β为主的免疫识别,该结果可为深入开展PHEV抑制机体干扰素产生的机制研究奠定基础。The porcine hemagglutinating encephalomyelitis virus （PHEV） is a beta coronavirus which invades the central nervous system（CNS）,in where nerve cells are a target for viral replication. Previous studies have shown that some coronaviruses could escape the innate immune surveil- lance of the host, resulting in obvious pathogenicity. In order to clarify the immune response of neurons in the process of PHEV infection,the important inflammatory cytokines and related path- way changes in innate immunity were analyzed. IFN-9 was tested in N2a cells infected with PHEV by ELISA at different time points. The result showed that PHEV could not induce IFN-β in N2a cells. When cell culture supernatants were probed for TNF-α, IL-1β, IL-6, IFN-γ protein by ELISA,only IL-6 showed high expression levels in N2a 24 hp. i. supernatant, the data indicated that PHEV infection could induce IL-6 innate immune response. Then we investigated the subcellular localization of IRF-3 and NF-κB which are important for inducing antiviral and inflammatory. NF-κB was transported from the cytoplasm to the nucleus, however, IRF-3 was still in nucleus,suggesting that PHEV only activated the NF-κB pathway. These results demonstrated that PHEV enters the central nervous system,evading nerve cells immune recognition mainly by IFN-β. It laid a foundation for further research on the mechanism of inhibiting the production of IFN.

关 键 词：猪血凝性脑脊髓炎病毒 干扰素 IL-6 NF-ΚB IRF-3 

分 类 号：S852[农业科学—基础兽医学]