Loss of furin site enhances SARS-CoV-2 spike protein pseudovirus infection.

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 (S_Fko) 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 S_Fko leads to an increased level of S protein and limits S1/S2 cleavage to enhance pseudovirus infection in cells. Full-length S (S_FL) pseudotyped with N, M, and E helper packaging can effectively help S_FL infect cells. Finally, pseudotyped S_Fko 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.

SUBMITTER: Wang Z 

PROVIDER: S-EPMC9790109 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature