Martin Ulrike, Nestler Matthias, Munder Thomas, Zell Roland, Sigusch Holger H, Henke Andreas
Institute of Virology and Antiviral Therapy, Medical Center, Friedrich Schiller University Jena, Winzerlaer Str. 10, 07745 Jena, Germany.
Med Microbiol Immunol. 2004 May;193(2-3):133-9. doi: 10.1007/s00430-003-0197-7. Epub 2003 Sep 12.
Among several mechanisms of pathogenesis of the frequent and sometimes serious infections with coxsackievirus B3 (CVB3), one detail is apoptosis. Recently, a new apoptotic mechanism involving the specific interaction between the capsid protein VP2 of the highly virulent variant CVB3H3 and the proapoptotic host protein Siva was identified. The relevance of this observation for virus pathogenicity was shown in a BALB/c mouse model using CVB3H3 and the interaction-deficient mutant virus CVB3H310A1. In this study these results were verified and extended under in vitro conditions. The different apoptotic capability of CVB3H3 versus CVB3H310A1 was demonstrated by apoptotic nuclear condensation, DNA fragmentation, expression of Siva mRNA, and caspase-3 activation. The virus-specific differences were caused by the VP2 capsid proteins, which was shown by overexpression of the single VP2H3 and VP2H310A1 protein. Furthermore, the involvement of apoptosis in virus progeny production and the associated appearance of the cytopathic effect was demonstrated by application of the pan-caspase inhibitor Z-VAD-FMK. These in vitro results indicate that the induction of apoptosis during CVB3H3 infection is based on the interaction between the capsid protein VP2 and the proapoptotic protein Siva, independently from the complex situation in vivo.
在柯萨奇病毒B3（CVB3）频繁且有时严重感染的几种发病机制中，一个细节是细胞凋亡。最近，一种涉及高毒力变异株CVB3H3的衣壳蛋白VP2与促凋亡宿主蛋白Siva之间特异性相互作用的新凋亡机制被发现。在使用CVB3H3和相互作用缺陷型突变病毒CVB3H310A1的BALB/c小鼠模型中，显示了这一观察结果与病毒致病性的相关性。在本研究中，这些结果在体外条件下得到了验证和扩展。通过凋亡核浓缩、DNA片段化、Siva mRNA表达和caspase-3激活，证明了CVB3H3与CVB3H310A1不同的凋亡能力。病毒特异性差异是由衣壳蛋白VP2引起的，单一VP2H3和VP2H310A1蛋白的过表达证明了这一点。此外，通过应用泛半胱天冬酶抑制剂Z-VAD-FMK，证明了细胞凋亡参与病毒子代产生以及相关细胞病变效应的出现。这些体外结果表明，CVB3H3感染期间细胞凋亡的诱导是基于衣壳蛋白VP2与促凋亡蛋白Siva之间的相互作用，与体内复杂情况无关。
