Overend Christopher C, Cui Junru, Grubman Marvin J, Garmendia Antonio E
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Rd, Storrs, CT, 06269, USA.
Department of Biomedical Sciences and Pathobiology Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1981 Kraft Drive, Blacksburg, VA, 24061-0913, USA.
Vet Res Commun. 2017 Mar;41(1):15-22. doi: 10.1007/s11259-016-9665-6. Epub 2016 Nov 28.
It has been recognized that the expression of type I interferon (IFNα/β) may be suppressed during infection with porcine reproductive, respiratory syndrome virus (PRRSV). This causes profound negative effects on both the innate and adaptive immunity of the host resulting in persistence of infection.
Test the effects of PRRSV infection of porcine alveolar macrophages (PAMs), the main target cell, on the expression of interferon beta (IFNβ) and downstream signaling events.
In order to examine those effects, PAMs harvested from lungs of healthy PRRSV-free animals were infected with virulent, attenuated, infectious clone-derived chimeric viruses, or field PRRS virus strains. Culture supernatants from the infected PAMs were tested for IFNβ protein expression by means of indirect ELISA and for bioactivity by a vesicular stomatitis virus plaque reduction assay. The expression of the Mx protein was assayed to ascertain signaling events.
These experiments demonstrated that PRRSV does induce variably, the expression of bioactive IFNβ protein in the natural host cell. To further elucidate the effects of PRRSV infection on IFNβ signaling, Mx-1 an interferon stimulated gene (ISG), was also tested for expression. Interestingly, Mx-1 expression by infected PAMs generally correlated with IFNβ production.
The results of this study demonstrate that the induction of IFNβ and signaling in PAMs after PRRSV infection is variable.
人们已经认识到,在感染猪繁殖与呼吸综合征病毒(PRRSV)期间,I型干扰素(IFNα/β)的表达可能会受到抑制。这会对宿主的固有免疫和适应性免疫产生深远的负面影响,导致感染持续存在。
检测PRRSV感染猪肺泡巨噬细胞(PAMs,主要靶细胞)对干扰素β(IFNβ)表达及下游信号事件的影响。
为了检测这些影响,从无PRRSV的健康动物肺中获取的PAMs用强毒株、弱毒株、感染性克隆衍生嵌合病毒或田间PRRS病毒株进行感染。通过间接ELISA检测感染的PAMs培养上清液中的IFNβ蛋白表达,并通过水疱性口炎病毒蚀斑减少试验检测其生物活性。检测Mx蛋白的表达以确定信号事件。
这些实验表明,PRRSV确实会在天然宿主细胞中不同程度地诱导生物活性IFNβ蛋白的表达。为了进一步阐明PRRSV感染对IFNβ信号传导的影响,还检测了干扰素刺激基因(ISG)Mx-1的表达。有趣的是,感染的PAMs中Mx-1的表达通常与IFNβ的产生相关。
本研究结果表明,PRRSV感染后PAMs中IFNβ的诱导和信号传导是可变的。