【摘要】：豬血凝性腦脊髓炎病毒(PHEV)屬于β屬冠狀病毒成員,其具有典型的嗜神經(jīng)性。多種冠狀病毒可逃避宿主先天性免疫的識(shí)別而表現(xiàn)出明顯的致病性,但宿主的先天性免疫在PHEV感染過(guò)程中發(fā)揮的作用還不是很清楚。為了明確神經(jīng)細(xì)胞在PHEV感染過(guò)程中細(xì)胞自身的免疫應(yīng)答狀況,本研究對(duì)參與先天性免疫的重要炎性細(xì)胞因子和相關(guān)的通路變化情況進(jìn)行了檢測(cè)分析。用酶聯(lián)免疫吸附試驗(yàn)(ELISA)方法對(duì)PHEV感染小鼠腦神經(jīng)瘤母細(xì)胞(N2a)不同時(shí)間點(diǎn)的樣品進(jìn)行檢測(cè),結(jié)果證明沒(méi)有IFN-β的表達(dá),說(shuō)明PHEV不能引起神經(jīng)細(xì)胞產(chǎn)生Ⅰ型IFN。收集感染N2a細(xì)胞24h的上清,ELISA方法檢測(cè)TNF-α、IL-1β、IL-6和IFN-γ的分泌情況,僅有IL-6呈現(xiàn)高水平表達(dá),說(shuō)明PHEV感染N2a細(xì)胞誘發(fā)了IL-6的先天性免疫反應(yīng)。免疫熒光方法檢測(cè)PHEV感染N2a細(xì)胞24h后重要核轉(zhuǎn)錄因子IRF-3和NF-κB的核定位情況,發(fā)現(xiàn)NF-κB移位到了胞核,IRF-3仍然存在于胞漿,說(shuō)明PHEV僅激活了NF-κB信號(hào)通路。以上試驗(yàn)結(jié)果表明,PHEV進(jìn)入中樞神經(jīng)系統(tǒng)時(shí)可逃避神經(jīng)細(xì)胞以IFN-β為主的免疫識(shí)別,該結(jié)果可為深入開(kāi)展PHEV抑制機(jī)體干擾素產(chǎn)生的機(jī)制研究奠定基礎(chǔ)。
[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- 偽, IL-1 尾, IL-6 and IFN- 緯 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.
【作者單位】： 吉林大學(xué)動(dòng)物醫(yī)學(xué)學(xué)院;四平梨樹(shù)縣動(dòng)物疫病預(yù)防控制中心;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(31472194,31602018) 吉林省青年科學(xué)基金資助項(xiàng)目(20160520033JH)
【分類(lèi)號(hào)】：S852.65
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本文編號(hào)：2363580