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Loss of furin site enhances SARS-CoV-2 spike protein pseudovirus infection.
Wang, Zeng; Zhong, Kunhong; Wang, Guoqing; Lu, Qizhong; Li, Hexian; Wu, Zhiguo; Zhang, Zongliang; Yang, Nian; Zheng, Meijun; Wang, Yuelong; Nie, Chunlai; Zhou, Liangxue; Tong, Aiping.
  • Wang Z; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Zhong K; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Wang G; Department of Neurosurgery, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Lu Q; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Li H; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Wu Z; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Zhang Z; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Yang N; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Zheng M; Department of Otolaryngology, Head and Neck Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Wang Y; Department of Neurosurgery, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, China.
  • Nie C; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
  • Zhou L; Department of Neurosurgery, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, China. Electronic address: liangxue_zhou@126.com.
  • Tong A; State Key Laboratory of Biotherapy and Cancer Center, Research Unit of Gene and Immunotherapy, Chinese Academy of Medical Sciences, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, China. Electronic address: aipingtong@scu.edu.cn.
Gene ; 856: 147144, 2023 Mar 10.
Article in English | MEDLINE | ID: covidwho-2165309
ABSTRACT

BACKGROUND:

SARS-CoV-2 has a significant impact on healthcare systems all around the world. Due to its high pathogenicity, live SARS-CoV-2 must be handled under biosafety level 3 conditions. Pseudoviruses are useful virological tools because of their safety and versatility, but the low titer of these viruses remains a limitation for their more comprehensive applications.

METHOD:

Here, we constructed a Luc/eGFP based on a pseudotyped lentiviral HIV-1 system to transduce SARS-CoV-2 S glycoprotein to detect cell entry properties and cellular tropism.

RESULTS:

The furin cleavage site deletion of the S protein removed (SFko) can help SARS-CoV-2 S to be cleaved during viral packaging to improve infection efficiency. The furin cleavage site in SARS-CoV-2-S mediates membrane fusion and SFko leads to an increased level of S protein and limits S1/S2 cleavage to enhance pseudovirus infection in cells. Full-length S (SFL) pseudotyped with N, M, and E helper packaging can effectively help SFL infect cells. Finally, pseudotyped SFko particles were successfully used to detect neutralizing antibodies in RBD protein-immunized mouse serum.

CONCLUSION:

Overall, our study indicates a series of modifications that result in the production of relatively high-titer SARS-COV-2 pseudo-particles that may be suitable for the detection of neutralizing antibodies from COVID-19 patients.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: COVID-19 Limits: Animals / Humans Language: English Journal: Gene Year: 2023 Document Type: Article Affiliation country: J.gene.2022.147144

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Full text: Available Collection: International databases Database: MEDLINE Main subject: COVID-19 Limits: Animals / Humans Language: English Journal: Gene Year: 2023 Document Type: Article Affiliation country: J.gene.2022.147144