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. 2021 Jun 30;12:666991. doi: 10.3389/fimmu.2021.666991

Figure 1.

Figure 1

Schematic of 7-day VIA. Following an overnight rest (Day 0), CD4 T cells isolated by negative bead selection (Day 1) are activated in bulk culture for 3 days. On day 4, CD4 T cells are infected with HIV-JRCSF by spin-oculation with addition of Polybrene™ and thawed CD8 T cells are isolated by positive bead selection from autologous PBMCs. Independent cultures of CD4 T cells not JRCSF infected (n = 1), JRCSF infected CD4 T cells (n = 6) and JRCSF-CD4 T cells + CD8 T cells in 1:2 (n = 6) are set up in round-bottom flow-cytometry tubes and then cultured for another 3 days. On Day 7, all cultures (13 per participant) are stained and acquired by flow cytometry to measure intracellular p24+ frequency in CD4 (CD3+CD8-) T cells. Flow data are exported to the VIA replicate calculator ( Supplementary Material 4 ) for automated calculation of weighted averages across replicate cultures, % virus inhibition and 95% confidence intervals.