猪血凝性脑脊髓炎病毒抑制神经细胞干扰素的产生
发布时间：2018-11-28 16:50
【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。
[Abstract]:Porcine hemagglutinating encephalomyelitis virus (PHEV) is a member of coronavirus and has typical neurophilic properties. Many coronaviruses can escape the recognition of host innate immunity and exhibit obvious pathogenicity, but the role of host innate immunity in the process of PHEV infection is not very clear. In order to determine the immune response of nerve cells in the process of PHEV infection, the changes of important inflammatory cytokines and related pathways involved in congenital immunity were detected and analyzed in this study. Enzyme linked immunosorbent assay (ELISA) was used to detect the samples of brain neuroblastocytes (N2a) from mice infected with PHEV at different time points. The results showed that there was no expression of IFN- 尾, indicating that PHEV could not induce neuronal cells to produce type I IFN.. The supernatant of N2a cells infected with PHEV for 24 hours was collected. The secretion of TNF- 伪, IL-1 尾, IL-6 and IFN- 纬 was detected by ELISA method. Only the high expression of IL-6 was found, which indicated that PHEV infection induced the innate immune response of IL-6. Immunofluorescence assay was used to detect the nuclear localization of IRF-3 and NF- 魏 B in N2a cells infected with PHEV for 24 hours. It was found that NF- 魏 B was translocated into the nucleus and IRF-3 was still present in the cytoplasm, indicating that PHEV only activated the NF- 魏 B signaling pathway. These results suggest that PHEV can escape the immunological recognition of IFN- 尾 when it enters the central nervous system, which may lay a foundation for further research on the mechanism of inhibition of interferon production by PHEV.
【作者单位】： 吉林大学动物医学学院;四平梨树县动物疫病预防控制中心;
【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)
【分类号】：S852.65
，
本文编号：2363579
[Abstract]:Porcine hemagglutinating encephalomyelitis virus (PHEV) is a member of coronavirus and has typical neurophilic properties. Many coronaviruses can escape the recognition of host innate immunity and exhibit obvious pathogenicity, but the role of host innate immunity in the process of PHEV infection is not very clear. In order to determine the immune response of nerve cells in the process of PHEV infection, the changes of important inflammatory cytokines and related pathways involved in congenital immunity were detected and analyzed in this study. Enzyme linked immunosorbent assay (ELISA) was used to detect the samples of brain neuroblastocytes (N2a) from mice infected with PHEV at different time points. The results showed that there was no expression of IFN- 尾, indicating that PHEV could not induce neuronal cells to produce type I IFN.. The supernatant of N2a cells infected with PHEV for 24 hours was collected. The secretion of TNF- 伪, IL-1 尾, IL-6 and IFN- 纬 was detected by ELISA method. Only the high expression of IL-6 was found, which indicated that PHEV infection induced the innate immune response of IL-6. Immunofluorescence assay was used to detect the nuclear localization of IRF-3 and NF- 魏 B in N2a cells infected with PHEV for 24 hours. It was found that NF- 魏 B was translocated into the nucleus and IRF-3 was still present in the cytoplasm, indicating that PHEV only activated the NF- 魏 B signaling pathway. These results suggest that PHEV can escape the immunological recognition of IFN- 尾 when it enters the central nervous system, which may lay a foundation for further research on the mechanism of inhibition of interferon production by PHEV.
【作者单位】： 吉林大学动物医学学院;四平梨树县动物疫病预防控制中心;
【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)
【分类号】：S852.65
，
本文编号：2363579
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