Fig. 3: HIV-1 upregulates ODC-1, EIF5A, EIF5A hypusination and polyamine synthesis in CD4⁺ T cells even after ARI treatment.

A HTOC were TCR-activated and infected with HIV. ARI was added the next day to block subsequent rounds of infection. Cells were allowed to expand in the presence of TGF-β1 (10 ng/ml) and IL-2 (100 U/ml) and ARI for 6 days as described in methods (n = 5 independent experiments). Representative flow cytometric data showing ODC-1 expression in CD3⁺CD8 negative cells (CD4+ T cells) (top) and statistical analyses of MFI from five independent experiments (bottom) are shown. B Key intermediate steps in polyamine synthesis. Cells were stimulated and processed for flow cytometry as in A for determining the expression of EIF5A (C) and hypusinated EIF5A in CD4⁺ T cells (D). Light and dark shaded gray histograms represent staining controls for EIF5A (C) and hypusinated EIF5A (D) respectively. Mean values +/− SEM; ****P < 0.0001; ***P < 0.0002; Two-tailed; Unpaired t test in A–D. E Fluorometric polyamine estimation in cellular lysates from cells stimulated as in A for 4 or 7 days (n = 3 independent experiments). Polyamine concentrations were normalized to cell numbers. Mean values +/− SD; ***P = 0.0002; Two-tailed; Welch’s t test. Circles within box plots represent each replicate.