TY  - JOUR
TI  - CD8 T Cell Virus Inhibition Assay Protocol
AU  - Xu, Yinyan 
AU  - Weideman, Ann Marie
AU  - Abad-Fernandez, Maria 
AU  - Mollan, Katie R.
AU  - Kallon, Sallay 
AU  - Samir, Shahryar 
AU  - Warren, Joanna A.
AU  - Clutton, Genevieve 
AU  - Roan, Nadia 
AU  - Adimora, Adaora A.
AU  - Archin, Nancie 
AU  - Kuruc, JoAnn 
AU  - Gay, Cindy 
AU  - Hudgens, Michael G.
AU  - Goonetilleke, Nilu 
VL  - 12
IS  - 6
PY  - 2022
DA  - 2022/03/20
SP  - e4354
C1  - Bio-protocol 2022;12:e4354
DO  - 10.21769/BioProtoc.4354
UR  - https://doi.org/10.21769/BioProtoc.4354
AB  - 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:
KW  - CD4
KW  - CD8
KW  - HIV
KW  - JRCSF
KW  - T-cell
KW  - p24
KW  - Virus inhibition
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False