Figure 5
HIV-1 induces inhibitory molecules and T-cell impairment through P38MAPK pathway. Mature DCs were pulsed with mock or HIV-1BaL, cultured for ∼24 h, washed and cocultured with TN cells for 8 d. (A) Intracellular staining for phosphorylated P38MAPK was performed after 8 d of culture to compare between HIV pulsed and mock DC primed T cells. P38MAPK inhibitor (500 nmol/L) (Sigma-SB220025) was added to the wells at the onset of coculture. The cocultures were restimulated on d 7 and (B) T-cell proliferation was measured for CFSE dilution by flow cytometry (n = 6). (C) The gene levels of CTLA-4, PD-1, TRAIL, TIM-3, CD160, LAG3, FOXP3, BLIMP-1 and DTX1 were assessed by qRT-PCR (n = 4). The experiments (A-C) were normalized, and statistical analysis was performed by one-way ANOVA (Tukey post hoc test). *P < 0.05, **P < 0.005, ***P < 0.001. (D) The protein profiles of CTLA-4, PD-1, TRAIL, TIM-3, CD160 and LAG3 were measured by flow cytometry (n = 4: one representative experiment shown).