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Author Spotlight: Studying Host-Virus Interactions with Pseudotyped Viruses
Published on: November 21, 2023
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Optimized Pseudotyping Conditions for the SARS-COV-2 Spike Glycoprotein
Marc C Johnson1, Terri D Lyddon2, Reinier Suarez2
1Department of Molecular Microbiology and Immunology, University of Missouri School of Medicine, Christopher S. Bond Life Sciences Center, Columbia, Missouri, USA marcjohnson@missouri.edu.
Journal of Virology
|August 14, 2020
Summary
Researchers optimized severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) Spike pseudotyped viruses for enhanced infectivity. These pseudotyped viruses are effective tools for detecting neutralizing antibodies against SARS-COV-2 in COVID-19 patient plasma.
Area of Science:
- Virology and Molecular Biology
- Immunology
- Biotechnology
Background:
- The SARS-COV-2 Spike glycoprotein mediates viral entry into host cells.
- Generating pseudotyped viral particles is crucial for studying viral infectivity and developing diagnostics.
- Optimizing Spike glycoprotein function in pseudotyped systems is essential for high-titer production.
Purpose of the Study:
- To characterize codon-optimized SARS-COV-2 Spike glycoprotein for pseudotyped virus production.
- To enhance the efficiency of pseudotyped virus generation using various modifications.
- To validate the utility of SARS-COV-2 pseudotyped viruses for detecting neutralizing antibodies.
Main Methods:
- Generation of pseudotyped HIV-1, MLV, and VSV particles using modified SARS-COV-2 Spike glycoproteins.
- Systematic deletion of the Spike protein's cytoplasmic tail and introduction of ACE2 and TMPRSS2.
- Evaluation of Spike protein signal peptide, D614G, and R682Q mutations for enhanced production.
- Titration of infectious pseudotyped particles and assessment of neutralizing antibody detection.
Main Results:
- Deletion of the C-terminal 19 amino acids of the Spike protein enhanced pseudotyped particle production over 10-fold.
- Co-expression of ACE2 and TMPRSS2 significantly increased target cell susceptibility to infection.
- Spike mutations D614G and R682Q further boosted infectious particle production.
- Combined modifications yielded HIV-1 pseudotyped particles with titers approaching 10^6 particles/ml.
- Pseudotyped HIV-1 particles successfully detected neutralizing antibodies in COVID-19 patient plasma.
Conclusions:
- Optimized SARS-COV-2 Spike glycoproteins enable high-titer production of pseudotyped viruses.
- These pseudotyped viral systems provide a sensitive and safe platform for detecting neutralizing antibodies.
- The developed method facilitates research on viral entry mechanisms and antibody-based diagnostics for SARS-COV-2.
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