Determination of neutralization activities by a new versatile assay using an HIV-1 genome carrying the Gaussia luciferase gene

Journal of Virological Methods
Yuepeng ZhangFeng Gao

Abstract

Characterization of neutralizing activities are critical to evaluation of the neutralization potency and breadth of monoclonal antibodies or anti-HIV-1 sera elicited during natural HIV-1 infection or by vaccines. We have developed a new neutralization method using the SG3Δenv genome carrying the Gaussia luciferase gene between the env and nef genes. Pseudotype viruses generated using this new SG3Δenv-GLuc backbone together with HIV-1 env genes were infectious to TZM-bl cells, T cell lines and primary T cells. Viral infection can be detected by measuring luciferase activities with both lysed cells and culture supernatants. Neutralization titers in sera from HIV-1-infected individuals against tier 1 and tier 2 viruses were comparable to those determined by the gold standard TZM-bl-firefly method. Since the neutralization activities can be determined by repeatedly measuring luciferase activities in culture supernatants of any cells that are infected by SG3Δenv-GLuc-Env pseudotype viruses, this new method can serve as a versatile and high throughput assay to determine neutralization activities.

Citations

Apr 25, 2020·Frontiers in Physiology·Madeleine Iafrate, Gilbert O Fruhwirth
Apr 4, 2020·Molecular Therapy : the Journal of the American Society of Gene Therapy·Candice Ashmore-HarrisGilbert O Fruhwirth

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