Fig. 2: Elevated IFNα effect on HIV coreceptor CCR5 expression in human CD4⁺T cells and on release of circulating HIV by infected CD4⁺T cells.

aI CCR5 mRNA expression levels, assessed by RT-qPCR, and (aII) CCR5 protein expression level, evaluated by flow cytometry in CD4⁺T cells stimulated with different concentrations of IFNα (mRNA analysis n = 4, protein analysis n = 7, purple) and IFNλ2 (mRNA analysis n = 5, protein analysis n = 4, orange). aIII CXCR4 mRNA expression levels assessed by RT-qPCR and (aIV) CXCR4 protein expression level evaluated by flow cytometry in CD4⁺T cells stimulated with different concentrations of IFNα (mRNA analysis n = 4, protein analysis n = 4). b Histograms show CCR5 frequency in CD4⁺T cells stimulated with different IFNα concentrations in each studied group (HDs n = 7, ECs n = 3, UPs n = 3, and TPs n = 5). Multiple group comparisons were made using the Kruskal–Wallis test with Dunn’s multiple comparison testing. Values are medians and p values (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001); ns: not significant. Error bars on graphs represent interquartile ranges. c PBMCs from a single HD were stimulated in the presence (pretreated) or absence (not pretreated) of IFNα for 4 days. The stimulated cells were infected by the CH058 T/F virus (CCR5 tropic) (cI) or the 40700 T/F virus (CXCR4-tropic) (cII). Upon infection, cells were cultured with the corresponding concentration of IFNα for 48 h. For the positive control, the infected cells (not pretreated) were cultured in the absence of IFNα for determination of the normal replication kinetics of the virus. The p24 concentration in the culture supernatants was measured at 2, 6, 12, 24, and 48 h post infection. The infections were performed in duplicate, and the error bar represents the standard deviation (SD). Experiments were performed in PBMCs from three HDs. The results of one representative experiment were shown. MFI: median fluorescence intensity.