African swine fever virus multigene family 360 and 530 genes affect host interferon response
African swine fever virus multigene family 360 and 530 genes affect host interferon response
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DOI：
10.1128/jvi.78.4.1858-1864.2004
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发表时间：
2004-02-01
影响因子：
5.4
通讯作者：
Rock, DL
中科院分区：
文献类型：
--
作者：
Afonso, CL;Piccone, ME;Rock, DL
非洲猪瘟病毒（ASFV）多基因家族360和530 （MGF360/530）基因对猪巨噬细胞培养病毒生长和毒力的影响[L. Zsak，卢志忠，T. G. Burrage， J. G. Neilan， G. F. Kutish， D. M. Moore， D. L. Rock， J.病毒学报，75:3066-3076,2001)。这些新基因影响病毒-宿主相互作用的机制尚不清楚。为了确定MGF360/530基因的功能，我们比较了感染亲代ASFV （Pr4）和MGF360/530缺失突变体（Pr4Delta35）后巨噬细胞的转录反应。利用含有7712个巨噬细胞cDNA克隆的猪cDNA芯片，比较了Pr4和Pr4Delta35感染后3和6小时（hpi）的猪巨噬细胞的转录谱。而在3 hpi时，大多数基因（7,564）在感染任一病毒的细胞中表达水平相似，38个基因的mRNA水平在感染pr4delta35的巨噬细胞中显著升高（bb0 2.0倍，P < 0.05）。这些基因在6 hpi时也出现了类似的上调。这种诱导的转录反应需要病毒感染。大多数Pr4Delta35上调的基因是I型干扰素（IFN）应答的一部分，或者是通常由双链RNA和/或病毒感染诱导的基因。这些蛋白包括单核细胞趋化蛋白、跨膜蛋白3、四肽重复蛋白1、泛素样17-kDa蛋白、泛素特异性蛋白酶ISG43、RNA解旋酶DEAD盒蛋白、gtp结合MX蛋白、细胞因子IP-10和PKR激活因子PACT。Northern blot和real-time PCR证实了Pr4Delta35中IFN早期反应基因相对于Pr4的差异表达。分析ifn - α mRNA和分泌的ifn - α在3,8和24 hpi时的水平显示，在模拟和pr4感染的巨噬细胞中检测不到ifn - α，但在pr4delta35感染的巨噬细胞中，在24 hpi时的ifn - α水平显著。pr4感染巨噬细胞中IFN- α的缺失表明MGF360/530基因直接或间接抑制I型IFN反应。无法抑制宿主I型IFN反应可能是猪巨噬细胞中Pr4Delta35生长缺陷及其衰减的原因。
African swine fever virus (ASFV) multigene family 360 and 530 (MGF360/530) genes affect viral growth in macrophage cell cultures and virulence in pigs (L. Zsak, Z. Lu, T. G. Burrage, J. G. Neilan, G. F. Kutish, D. M. Moore, and D. L. Rock, J. Virol. 75:3066-3076, 2001). The mechanism by which these novel genes affect virus-host interactions is unknown. To define MGF360/530 gene function, we compared macrophage transcriptional responses following infection with parental ASFV (Pr4) and an MGF360/530 deletion mutant (Pr4Delta35). A swine cDNA microarray containing 7,712 macrophage cDNA clones was used to compare the transcriptional profiles of swine macrophages infected with Pr4 and Pr4Delta35 at 3 and 6 h postinfection (hpi). While at 3 hpi most (7,564) of the genes had similar expression levels in cells infected with either virus, 38 genes had significantly increased (> 2.0-fold, P < 0.05) mRNA levels in Pr4Delta35-infected macrophages. Similar up-regulation of these genes was observed at 6 hpi. Viral infection was required for this induced transcriptional response. Most Pr4Delta35 up-regulated genes were part of a type I interferon (IFN) response or were genes that are normally induced by double-stranded RNA and/or viral infection. These included monocyte chemoattractant protein, transmembrane protein 3, tetratricopeptide repeat protein 1, a ubiquitin-like 17-kDa protein, ubiquitin-specific protease ISG43, an RNA helicase DEAD box protein, GTP-binding MX protein, the cytokine IP-10, and the PKR activator PACT. Differential expression of IFN early-response genes in Pr4Delta35 relative to Pr4 was confirmed by Northern blot analysis and real-time PCR. Analysis of IFN-alpha mRNA and secreted IFN-alpha levels at 3, 8, and 24 hpi revealed undetectable IFN-alpha in mock- and Pr4-infected macrophages but significant IFN-alpha levels at 24 hpi in Pr4Delta35-infected macrophages. The absence of IFN-alpha in Pr4-infected macrophages suggests that MGF360/530 genes either directly or indirectly suppress a type I IFN response. An inability to suppress host type I IFN responses may account for the growth defect of Pr4Delta35 in macrophages and its attenuation in swine.