Figure 6.

Inhibition of HIV Nef interaction with ASK1 in the CD4 T cells procured from patients on ART results in increased susceptibility to killing by autologous CD8 T cells. CD4 T cells and autologous CD8 T cells were procured from the PBMC of four HIV-infected patients on ART and matched healthy controls. The CD4 T cells were incubated with ASK1 peptide that interferes with Nef-ASK1 interaction. Following the incubation with or without the peptide, the CD4 T cells were mixed with autologous CD8 T cells for the cytotoxic assay. The cells were then fixed and labeled with anti-CD4 antibody, anti-CD8 antibody, anti-active caspase-3 antibody and DNA fragmentation assay (TUNEL assay) and analyzed by the ImageStream. (A) Four independent conjugation and apoptosis assays are presented. ** indicate P-values < 0.01, as obtained by two-tailed paired t-test. (B) Representative imaging flow cytometry plots of CD4 T cells incubated with the peptide and interacted with CD8 T cells. (C) ImageStream images of TUNEL and activated caspase-3 positive CD4 T cells treated with ASK1 peptide conjugated with autologous CD8 T cells.