Journal of Mazandaran University of Medical Sciences (Nov 2023)
Design and Development of a Pseudotyped-lentivirus for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
Abstract
Background and purpose: Pseudoviruses, due to their immunity and adaptability, are valuable tools for developing vaccines and therapeutic drugs, particularly for emerging viruses. SARS-CoV-2, due to its high mortality rate and ease of transmission, should be handled under Biosafety Level 3 conditions, making the development of vaccines and therapeutic drugs challenging in many research centers. The present study aimed to produce a lentivirus pseudotype that can be used in neutralization assays based on pseudovirus under Biosafety Level 2 conditions. Materials and methods: The SARS-CoV-2 Spike protein gene was inserted into the HIV-1 genome. Confirmation of entry was obtained through flow cytometry, electron microscopy, and immunofluorescence. Finally, the neutralization capacity of convalescent COVID-19 patients' sera was measured using the pseudovirus-based neutralization assay. Results: The results of flow cytometry, electron microscopy, and immunofluorescence confirmed the entry of the Spike gene. Furthermore, the sera of convalescent patients showed significantly higher neutralizing capacity compared to the control group that had not been exposed to SARS-CoV-2. Conclusion: The results of this study demonstrated that the use of the produced lentivirus pseudotype can measure the neutralizing capacity of patients' sera, and the pseudovirus-based neutralization assay serves as a suitable alternative for assessing the neutralization of live SARS-CoV-2. Moreover, this test can effectively evaluate vaccines or therapeutic drugs for combating this deadly virus.
