Messud-Petit F, Gelfi J, Delverdier M, Amardeilh M F, Py R, Sutter G, Bertagnoli S
Laboratoire Associe de Microbiologie Moleculaire, Institut National de la Recherche Agronomique and Ecole Nationale Vétérinaire, Toulouse, France.
J Virol. 1998 Oct;72(10):7830-9. doi: 10.1128/JVI.72.10.7830-7839.1998.
Recently, myxoma virus was shown to encode an additional member of the serpin superfamily. The viral gene, called serp2, was cloned, and the Serp2 protein was shown to specifically bind to interleukin-1beta (IL-1beta)-converting enzyme (ICE), thus inhibiting the cleavage of pro-IL-1beta by the protease (F. Petit, S. Bertagnoli, J. Gelfi, F. Fassy, C. Boucraut-Baralon, and A. Milon, J. Virol. 70:5860-5866, 1996). Here, we address the role of Serp2 in the development of myxomatosis, a lethal infectious disease of the European rabbit. A Serp2 mutant myxoma virus was constructed by disruption of the single-copy serp2 gene and insertion of the Escherichia coli gpt gene serving as the selectable marker. A revertant virus was obtained by replacing the E. coli gpt gene by the intact serp2 open reading frame. The Serp2(-) mutant virus replicated with wild-type kinetics both in rabbit fibroblasts and a rabbit CD4(+) T-cell line (RL5). Moderate reduction of cell surface levels of major histocompatibility complex I was observed after infection with wild-type or Serp2(-) mutant myxoma virus, and both produced white pocks on the chorioallantoic membrane of the chick embryo. After the infection of European rabbits, the Serp2(-) mutant virus proved to be highly attenuated compared to wild-type myxoma virus, as demonstrated by the clinical course of myxomatosis and the survival rates of infected animals. Pathohistological examinations revealed that infection with wild-type myxoma virus resulted in a blockade of the inflammatory response at the vascular level. In contrast, rapid inflammatory reactions occurred upon infection with the Serp2(-) mutant virus. Furthermore, lymphocytes in lymph nodes derived from animals inoculated with Serp2 mutant virus were shown to rapidly undergo apoptosis. We postulate that the virulence of myxoma virus in the European rabbit can be partially attributed to an impairment of host inflammatory processes and to the prevention of apoptosis in lymphocytes. The weakening of host defense is directly linked to serp2 gene function and is likely to involve the inhibition of IL-1beta-converting-enzyme-dependent pathways.
最近,黏液瘤病毒被证明编码丝氨酸蛋白酶抑制剂(serpin)超家族的另一个成员。这个名为serp2的病毒基因被克隆出来,并且Serp2蛋白被证明能特异性结合白细胞介素-1β(IL-1β)转化酶(ICE),从而抑制蛋白酶对前体IL-1β的切割(F. Petit、S. Bertagnoli、J. Gelfi、F. Fassy、C. Boucraut-Baralon和A. Milon,《病毒学杂志》70:5860 - 5866,1996年)。在此,我们探讨Serp2在兔黏液瘤病(一种欧洲兔的致死性传染病)发展过程中的作用。通过破坏单拷贝的serp2基因并插入用作选择标记的大肠杆菌gpt基因,构建了一种Serp2突变黏液瘤病毒。通过用完整的serp2开放阅读框取代大肠杆菌gpt基因获得了一种回复病毒。Serp2(-)突变病毒在兔成纤维细胞和兔CD4(+)T细胞系(RL5)中均以野生型动力学进行复制。在用野生型或Serp2(-)突变黏液瘤病毒感染后,观察到主要组织相容性复合体I的细胞表面水平有适度降低,并且两者在鸡胚绒毛尿囊膜上都产生白色痘疱。在感染欧洲兔后,与野生型黏液瘤病毒相比,Serp2(-)突变病毒被证明高度减毒,这在黏液瘤病的临床病程和感染动物的存活率中得到了证明。病理组织学检查显示,野生型黏液瘤病毒感染导致血管水平的炎症反应受阻。相反,用Serp2(-)突变病毒感染后会迅速发生炎症反应。此外,接种Serp2突变病毒的动物淋巴结中的淋巴细胞显示出迅速发生凋亡。我们推测,黏液瘤病毒在欧洲兔中的毒力部分可归因于宿主炎症过程的受损以及淋巴细胞凋亡的预防。宿主防御的减弱与serp2基因功能直接相关,并且可能涉及对IL-1β转化酶依赖性途径的抑制。