Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Ann Weideman
Department of Biostatistics, UNC Chapel Hill, Chapel Hill, North Carolina, USACenter for AIDS Research, School of Medicine, UNC Chapel Hill, Chapel Hill, NC 27599, USA
Maria Abad-Fernandez
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Katie Mollan
Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, USASchool of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Sallay Kallon
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Shahryar Samir
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Joanna Warren
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Genevieve Clutton
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Nadia Roan
Department of Urology, University of California San Francisco, CA 94158, USAGladstone Institute of Virology and Immunology, CA 94158, USA
Adaora Adimora
Center for AIDS Research, School of Medicine, UNC Chapel Hill, Chapel Hill, NC 27599, USADepartment of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA, School of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Nancie Archin
School of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
JoAnn Kuruc
School of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Cindy Gay
School of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
Michael Hudgens
Department of Biostatistics, UNC Chapel Hill, Chapel Hill, North Carolina, USA, Center for AIDS Research, School of Medicine, UNC Chapel Hill, Chapel Hill, NC 27599, USA
Nilu Goonetilleke
Department of Microbiology & Immunology, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USASchool of Medicine and UNC HIV Cure Center, UNC Chapel Hill School of Medicine, Chapel Hill, North Carolina, USA
The human immunodeficiency virus (HIV)-1 viral inhibition assay (VIA) measures CD8+ T cell-mediated inhibition of HIV replication in CD4+ T cells and is increasingly used for clinical testing of HIV vaccines and immunotherapies. Different VIAs that differ in length of CD8:CD4 T cell culture periods (6–13 days), purity of CD4 cultures [isolated CD4+ T cells or CD8+ depleted peripheral blood mononuclear cells (PBMCs)], HIV strains (laboratory strains, isolates, reporter viruses) and read-outs of virus inhibition (p24 ELISA, intracellular measurement of p24, luciferase reporter expression, and viral gag RNA) have been reported.Here, we describe multiple modifications to a 7-day VIA protocol, the most impactful being the introduction of independent replicate cultures for both HIV infected-CD4 (HIV-CD4) and HIV-CD4:CD8 T cell cultures. Virus inhibition was quantified using a ratio of weighted averages of p24+ cells in replicate cultures and the corresponding 95% confidence intervals. We identify methodological and analysis changes that could be incorporated into other protocols to improve assay reproducibility. We found that in people living with HIV (PLWH) on antiretroviral therapy (ART), CD8 T cell virus inhibition was largely stable over time, supporting the use of this assay and/or analysis methods to examine therapeutic interventions.Graphic abstract:
