禽呼肠孤病毒感染对SPF鸡外周血淋巴细胞中天然免疫相关基因转录的影响

Effects of Avian Reovirus Infection on Transcription of Innate Immune Genes in Peripheral Blood Lymphocytes of SPF Chickens

为了明确禽呼肠孤病毒(avian reovirus,ARV)感染对宿主天然免疫应答的影响,利用实时荧光定量PCR方法检测ARV感染SPF鸡后外周血淋巴细胞中天然免疫相关基因,包括Toll样受体(TLRs)、MDA5、IPS-1、IRF-3、IFN-α、IFN-β、IFN-γ、IFITM3、Mx1和OASL等基因在感染后0、12h以及1、2、3、5和7d的转录水平变化。结果表明,ARV感染SPF鸡后7d以内,外周血淋巴细胞中TLR3和TLR21的mRNA转录水平显著降低(P<0.05或P<0.01);而TLR7和MDA5的mRNA转录水平则在整个试验过程中显著升高,分别在7dpi和1dpi达到峰值(P<0.05或P<0.01)。IPS-1的mRNA转录被抑制;IRF-3则呈显著转录上调趋势;IFN-α、IFN-β和IFN-γ均表达上调,分别在2~3dpi达到峰值(P<0.05或P<0.01)。IFITM3、Mx1、OASL的mRNA转录水平也显著升高,均在3dpi达到峰值(P<0.01)。试验结果表明ARV在感染早期能够显著性引起TLR7、MDA5、IRF3、IFN-α、IFN-β、IFN-γ、IFITM3、Mx1和OASL的转录上调,抑制TLR3、TLR21和IPS-1的转录,为进一步阐释ARV的分子致病机制和免疫应答机制奠定了基础。

This study was aimed to assess the influences of avian reovirus（ARV）infection on expression of innate immune genes in peripheral blood lymphocytes of SPF chickens.The quantitative real-time PCR assays was performed to determine the transcriptional levels of TLR3,TLR7,TLR21,MDA5,IPS-1,IRF-3,IFN-α,IFN-β,IFN-γ,IFITM3,Mx1 and OASLin 0h,12 h,1,2,3,5and 7days post infection（dpi）in peripheral blood lymphocytes of SPF chickens infected with ARV S1133.These results showed that the mRNA transcriptional levels of TLR3 and TLR21were significantly reduced within 7dpi（P〈0.05 or P〈0.01）;the mRNA transcriptional levels of TLR7 and MDA5were obviously promoted and specially reached the summit maximumvalues on 7dpi and 1dpi,respectively（P〈0.05 or P〈0.01）.The mRNA level of IPS-1was down-regulated within 7dpi.Meanwhile,the mRNA level of IRF-3was up-regulated during the whole transcrption course,and IFN-α,IFN-βand IFN-γ mRNA levels were significantly increased and reached the peak at 2-3dpi.The mRNA levels of IFITM3,Mx1 and OASLwere significantly up-regulated and peaked at 3dpi（P〈0.01）.Therefore,ARV infection is able to modulate the increased mRNA transcription of TLR7,MDA5,IRF-3,IFN-α,IFN-β,IFN-γ,IFITM3,Mx1 and OASL,and reduce the mRNA transcription of TLR3,TLR21 and IPS-1,which would contribute to further understand ARV pathogenesis and host innate immune responses to ARV infection.