Skip to main content
. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: J Immunol. 2018 Jun 22;201(3):971–981. doi: 10.4049/jimmunol.1701551

Figure 1. Stimulation of PBMCs by HIV Gag induces a CD4 T cell-dependent production of cytokines by NK cells.

Figure 1.

PBMCs were stimulated in vitro with an HIV Gag peptide pool or left unstimulated (negative control) and assessed by a delayed ICS assay, in which cells were incubated with Ag for 12–36 before addition of brefeldin and monensin for 12h and collection after 24–48h. The flow cytometry plots show IFNγ production by CD4 T cells (CD3+CD4+), B cells (CD19+), NK cells (CD56+) and monocytes (CD14+) after doublet exclusion on FSC-A/FSC-H and dead cell exclusion. (A) Example plots for one representative donor showing IFN-γ production by CD4 T cells (CD3+CD4+), B cells (CD3-CD19+), NK cells (CD3-CD56+) and CD14+ monocytes, after doublet exclusion on FSC-A/FSC-H and dead cell exclusion after 24 or 48h of stimulation. (B) ICS frequencies of IFN-γ-producing NK cells 48h after Gag stimulation of PBMCs (n=21 untreated subjects) correlated (C) with HIV viral loads; and (D) with HIV Gag-specific CD4 T cell responses. (E) Representative example and (F) summary data on 7 subjects of the impact of CD3+ or CD8+ cell depletion on IFN-γ production by NK cells. Statistical tests: Wilcoxon paired test (BF) and Spearman correlation (CD).