Program Infection and Cancer, Division Tumor Virology F010, and Institut National de la Santé et de la Recherche Médicale U701, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
J Virol. 2012 Jul;86(14):7554-64. doi: 10.1128/JVI.00212-12. Epub 2012 May 2.
An in-frame, 114-nucleotide-long deletion that affects the NS-coding sequence was created in the infectious molecular clone of the standard parvovirus H-1PV, thereby generating Del H-1PV. The plasmid was transfected and further propagated in permissive human cell lines in order to analyze the effects of the deletion on virus fitness. Our results show key benefits of this deletion, as Del H-1PV proved to exhibit (i) higher infectivity (lower particle-to-infectivity ratio) in vitro and (ii) enhanced tumor growth suppression in vivo compared to wild-type H-1PV. This increased infectivity correlated with an accelerated egress of Del H-1PV progeny virions in producer cells and with an overall stimulation of the viral life cycle in subsequently infected cells. Indeed, virus adsorption and internalization were significantly improved with Del H-1PV, which may account for the earlier appearance of viral DNA replicative forms that was observed with Del H-1PV than wild-type H-1PV. We hypothesize that the internal deletion within the NS2 and/or NS1 protein expressed by Del H-1PV results in the stimulation of some step(s) of the viral life cycle, in particular, a maturation step(s), leading to more efficient nuclear export of infectious viral particles and increased fitness of the virus produced.
在标准细小病毒 H-1PV 的感染性分子克隆中创建了一个影响 NS 编码序列的 114 个核苷酸长的框内缺失,从而产生了 Del H-1PV。该质粒被转染,并在允许的人细胞系中进一步繁殖,以分析缺失对病毒适应性的影响。我们的结果表明了这种缺失的关键益处,因为与野生型 H-1PV 相比,Del H-1PV 表现出(i)更高的体外感染性(更低的颗粒/感染性比)和(ii)更强的体内肿瘤生长抑制作用。这种增强的感染性与 Del H-1PV 产生的病毒粒子在产生细胞中的更快释放有关,并且与随后感染的细胞中的病毒生命周期的整体刺激有关。事实上,Del H-1PV 的病毒吸附和内化显著改善,这可能解释了与野生型 H-1PV 相比,Del H-1PV 更早出现的病毒 DNA 复制形式。我们假设 Del H-1PV 表达的 NS2 和/或 NS1 蛋白中的内部缺失导致病毒生命周期的某些步骤(特别是成熟步骤)被刺激,从而导致更有效的核输出感染性病毒颗粒和产生的病毒适应性增强。