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

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.

Keywords: Gaussia luciferase; HIV-1; Neutralization; Pseudotype viruses.