Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
J Med Virol. 2021 Dec;93(12):6671-6685. doi: 10.1002/jmv.27244. Epub 2021 Aug 4.
Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a wide spectrum of syndromes involving multiple organ systems and is primarily mediated by viral spike (S) glycoprotein through the receptor-binding domain (RBD) and numerous cellular proteins including ACE2, transmembrane serine protease 2 (TMPRSS2), kidney injury molecule-1 (Kim-1), and neuropilin-1 (NRP-1). In this study, we examined the entry tropism of SARS-CoV-2 and SARS-CoV using S protein-based pseudoviruses to infect 22 cell lines and 3 types of primary cells isolated from respiratory, urinary, digestive, reproductive, and immune systems. At least one cell line or type of primary cell from each organ system was infected by both pseudoviruses. Infection by pseudoviruses is effectively blocked by S1, RBD, and ACE2 recombinant proteins, and more weakly by Kim-1 and NRP-1 recombinant proteins. Furthermore, cells with robust SARS-CoV-2 pseudovirus infection had strong expression of either ACE2 or Kim-1 and NRP-1 proteins. ACE2 glycosylation appeared to be critical for the infections of both viruses as there was a positive correlation between infectivity of either SARS-CoV-2 or SARS-CoV pseudovirus with the level of glycosylated ACE2 (gly-ACE2). These results reveal that SARS-CoV-2 cell entry could be mediated by either an ACE2-dependent or -independent mechanism, thus providing a likely molecular basis for its broad tropism for a wide variety of cell types.
严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)感染可引起涉及多个器官系统的广泛综合征,主要由病毒刺突(S)糖蛋白通过受体结合域(RBD)和许多细胞蛋白介导,包括 ACE2、跨膜丝氨酸蛋白酶 2(TMPRSS2)、肾损伤分子-1(Kim-1)和神经纤毛蛋白-1(NRP-1)。在这项研究中,我们使用基于 S 蛋白的假病毒来检测 SARS-CoV-2 和 SARS-CoV 的进入嗜性,以感染 22 种细胞系和 3 种从呼吸道、泌尿、消化、生殖和免疫系统分离的原代细胞。至少有一种来自每个器官系统的细胞系或原代细胞被两种假病毒感染。S1、RBD 和 ACE2 重组蛋白可有效阻断假病毒感染,而 Kim-1 和 NRP-1 重组蛋白的阻断效果较弱。此外,具有强大 SARS-CoV-2 假病毒感染的细胞强烈表达 ACE2 或 Kim-1 和 NRP-1 蛋白。ACE2 糖基化似乎对两种病毒的感染至关重要,因为 SARS-CoV-2 或 SARS-CoV 假病毒的感染性与糖基化 ACE2(gly-ACE2)水平呈正相关。这些结果表明,SARS-CoV-2 细胞进入可能通过 ACE2 依赖或独立的机制介导,从而为其广泛的对各种细胞类型的嗜性提供了可能的分子基础。