Department of Pediatrics, and the Elizabeth B. Lamb Center for Pediatric Research, D7235 Medical Center North, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Proc Natl Acad Sci U S A. 2009 Nov 24;106(47):19986-91. doi: 10.1073/pnas.0907412106. Epub 2009 Nov 6.
Serotype-specific patterns of reovirus disease in the CNS of newborn mice segregate with the viral S1 gene segment, which encodes attachment protein sigma1 and nonstructural protein sigma1s. The importance of receptor recognition in target cell selection by reovirus implicates the sigma1 protein as the primary effector of disease outcome. However, the contribution of sigma1s to reovirus disease is unknown. To define the function of sigma1s in reovirus pathogenesis, we generated a sigma1s-deficient virus by altering a single nucleotide to disrupt the sigma1s translational start site. Viruses were recovered that contain nine gene segments from strain type 3 Dearing and either the wild-type or sigma1s-null S1 gene segment from strain type 1 Lang. Following peroral inoculation of newborn mice, both viruses replicated in the intestine, although the wild-type virus achieved higher yields than the sigma1s-null virus. However, unlike the wild-type virus, the sigma1s-deficient virus failed to disseminate to sites of secondary viral replication, including the brain, heart, and liver. Within the small intestine, both viruses were detected in Peyer's patches, but only the wild-type virus reached the mesenteric lymph node. Concordantly, wild-type virus, but not sigma1s-deficient virus, was detected in the blood of infected animals. Wild-type and sigma1s-null viruses produced equivalent titers following intracranial inoculation, indicating that sigma1s is dispensable for viral growth in the murine CNS. These results suggest a key role for sigma1s in virus spread from intestinal lymphatics to the bloodstream, thereby allowing the establishment of viremia and dissemination to sites of secondary replication within the infected host.
呼肠孤病毒在新生鼠中枢神经系统中的血清型特异性疾病模式与编码附着蛋白 σ1 和非结构蛋白 σ1s 的病毒 S1 基因片段分离。在呼肠孤病毒靶细胞选择中,受体识别的重要性暗示 σ1 蛋白是疾病结果的主要效应因子。然而,σ1s 对呼肠孤病毒疾病的贡献尚不清楚。为了确定 σ1s 在呼肠孤病毒发病机制中的功能,我们通过改变一个核苷酸来破坏 σ1s 的翻译起始位点,从而产生了一种 σ1s 缺失病毒。从 3 型 Dearing 株中回收了包含 9 个基因片段的病毒,并且从 1 型 Lang 株中回收了含有野生型或 σ1s 缺失的 S1 基因片段的病毒。在对新生鼠进行口服接种后,两种病毒都在肠道中复制,尽管野生型病毒的产量高于 σ1s 缺失病毒。然而,与野生型病毒不同的是,σ1s 缺失病毒未能传播到二次病毒复制部位,包括大脑、心脏和肝脏。在小肠中,两种病毒都在派尔集合淋巴结中检测到,但只有野生型病毒到达肠系膜淋巴结。一致地,只有野生型病毒而不是 σ1s 缺失病毒在感染动物的血液中被检测到。野生型和 σ1s 缺失病毒在颅内接种后产生了相当的滴度,表明 σ1s 对于病毒在小鼠中枢神经系统中的生长是可有可无的。这些结果表明 σ1s 在病毒从肠道淋巴液传播到血液中起着关键作用,从而允许建立病毒血症和在感染宿主内传播到二次复制部位。