Mechanisms of zoonotic severe acute respiratory syndrome coronavirus host range expansion in human airway epithelium
Mechanisms of zoonotic severe acute respiratory syndrome coronavirus host range expansion in human airway epithelium
复制标题
DOI：
10.1128/jvi.02041-07
复制
发表时间：
2008-03-01
影响因子：
5.4
通讯作者：
Baric, Ralph
中科院分区：
文献类型：
--
作者：
Sheahan, Timothy;Rockx, Barry;Baric, Ralph
2003年，严重急性呼吸综合征冠状病毒（SARS-CoV）出现，在全球范围内造成8,000多例人类感染病例和700多人死亡。人畜共患的 SARS-CoV 很可能通过在中国销售的一系列人与动物之间的传播事件而进化为感染人类。利用合成生物学，我们将果子狸毒株 SZ16 的刺突蛋白 (S) 工程化到我们的流行毒株感染性克隆中，创建了嵌合病毒 icSZ16-S，该病毒具有感染性，但产生的后代病毒无法在体外繁殖。在 SZ16 的 S 受体结合域 (RBD) 内引入 K479N 突变后，重组病毒 (icSZ16-SK479N) 在 Vero 细胞中复制，但生长严重衰弱。 icSZ16-S K479N 在人气道上皮 (HAE) 细胞上的体外进化产生了两种病毒 (icSZ16-S K479N D8 和 D22)，它们在 HAE 细胞和表达 SARS-CoV 受体、人血管紧张素 I 转换酶 2 (hACE2) 的延迟性脑肿瘤细胞上生长增强。 icSZ16-S K479N D8 和 D22 病毒 RBD 包含 ACE2 接触残基 Y442F 和 L472F 的突变，从而重塑了 S 与 hACE2 的相互作用。此外，这些病毒被人单克隆抗体 (MAb) S230.15 中和，但亲本 icSZ16-SK479N 菌株的耐药性是突变体的八倍。这些数据表明，人畜共患 SARS-CoV 毒株的人类适应可能会选择一些对与 RBD 结合的单克隆抗体高度敏感的变体。流行病 icSZ16-S K479N 和 icSZ16-S K479N D22 病毒在 BALB/c 小鼠肺中复制相似，突显了这些人畜共患尖峰 SARS-CoV 在体内评估疫苗或血清疗法功效的潜在用途。
In 2003, severe acute respiratory syndrome coronavirus (SARS-CoV) emerged and caused over 8,000 human cases of infection and more than 700 deaths worldwide. Zoonotic SARS-CoV likely evolved to infect humans by a series of transmission events between humans and animals for sale in China. Using synthetic biology, we engineered the spike protein (S) from a civet strain, SZ16, into our epidemic strain infectious clone, creating the chimeric virus icSZ16-S, which was infectious but yielded progeny viruses incapable of propagating in vitro. After introducing a K479N mutation within the S receptor binding domain (RBD) of SZ16, the recombinant virus (icSZ16-S K479N) replicated in Vero cells but was severely debilitated in growth. The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells and on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2). The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2. Further, these viruses were neutralized by a human monoclonal antibody (MAb), S230.15, but the parent icSZ16-S K479N strain was eight times more resistant than the mutants. These data suggest that the human adaptation of zoonotic SARS-CoV strains may select for some variants that are highly susceptible to select MAbs that bind to RBDs. The epidemic, icSZ16-S K479N, and icSZ16-S K479N D22 viruses replicate similarly in the BALB/c mouse lung, highlighting the potential use of these zoonotic spike SARS-CoVs to assess vaccine or serotherapy efficacy in vivo.