National Center for International Research of Bio-targeting Theranostics, Guangxi Key Laboratory of Bio-targeting Theranostics, Collaborative Innovation Center for Targeting Tumor Diagnosis and Therapy, Guangxi Talent Highland of Bio-targeting Theranostics, Guangxi Medical University, Nanning, People's Republic of China.
National Translational Science Center for Molecular Medicine & Department of Cell Biology, Fourth Military Medical University, Xi'an, People's Republic of China.
Emerg Microbes Infect. 2022 Dec;11(1):1135-1144. doi: 10.1080/22221751.2022.2059403.
The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants is threatening public health around the world. Endocytosis functions as an important way for viral infection, and SARS-CoV-2 bears no exception. However, the specific endocytic mechanism of SARS-CoV-2 remains unknown. In this study, we used endocytic inhibitors to evaluate the role of different endocytic routes in SARS-CoV-2 pseudovirus infection and found that the viral infection was associated with caveolar/lipid raft- and cytoskeleton-mediated endocytosis, but independent of the clathrin-mediated endocytosis and macropinocytosis. Meanwhile, the knockdown of CD147 and Rab5a in Vero E6 and Huh-7 cells inhibited SARS-CoV-2 pseudovirus infection, and the co-localization of spike protein, CD147, and Rab5a was observed in pseudovirus-infected Vero E6 cells, which was weakened by CD147 silencing, illustrating that SARS-CoV-2 pseudovirus entered the host cells via CD147-mediated endocytosis. Additionally, Arf6 silencing markedly inhibited pseudovirus infection in Vero E6 and Huh-7 cells, while little change was observed in CD147 knockout-Vero E6 cells. This finding indicated Arf6-mediated CD147 trafficking plays a vital role in SARS-CoV-2 entry. Taken together, our findings provide new insights into the CD147-Arf6 axis in mediating SARS-CoV-2 pseudovirus entry into the host cells, and further suggest that blockade of this pathway seems to be a feasible approach to prevent the SARS-CoV-2 infection clinically.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）及其变体的传播正在威胁着全球公共卫生。内吞作用是病毒感染的一种重要方式，SARS-CoV-2 也不例外。然而，SARS-CoV-2 的具体内吞机制尚不清楚。在本研究中，我们使用内吞抑制剂来评估不同内吞途径在 SARS-CoV-2 假病毒感染中的作用，发现病毒感染与网格蛋白/脂筏和细胞骨架介导的内吞作用有关，但不依赖于网格蛋白介导的内吞作用和胞饮作用。同时，在 Vero E6 和 Huh-7 细胞中敲低 CD147 和 Rab5a 抑制了 SARS-CoV-2 假病毒感染，并且在假病毒感染的 Vero E6 细胞中观察到刺突蛋白、CD147 和 Rab5a 的共定位，这种共定位在 CD147 沉默后被削弱，表明 SARS-CoV-2 假病毒通过 CD147 介导的内吞作用进入宿主细胞。此外，Arf6 沉默显著抑制了 Vero E6 和 Huh-7 细胞中的假病毒感染，而在 CD147 敲除-Vero E6 细胞中几乎没有观察到变化。这一发现表明 Arf6 介导的 CD147 转运在 SARS-CoV-2 进入宿主细胞中起着至关重要的作用。综上所述，我们的研究结果为 CD147-Arf6 轴在介导 SARS-CoV-2 假病毒进入宿主细胞提供了新的见解，并进一步表明阻断该途径似乎是预防 SARS-CoV-2 感染的一种可行方法。
