Fig. 2

Clofarabine induced depletion of cellular dNTPs and inhibition of reverse transcription. Effect of clofarabine on cellular dNTP levels in primary activated CD4⁺ T cells (a) and macrophages (b) from three healthy donors. Activated CD4⁺ T cells and macrophages were treated with the indicated concentration of clofarabine for 8 h, washed with PBS, and dNTPs were extracted for analysis. dNTP levels are expressed as a percentage of the vehicle control (DMSO). Each value represents an individual donor with mean ± SD indicated. *p < .05 compared to vehicle control (multiple t test with Holm-Sidak post hoc test). c Cellular dATP concentrations of untreated and clofarabine (300 nM) treated T cells and macrophages from three healthy donors. Measured dATP levels were converted to cellular dATP concentrations as previously described [8], and are expressed as mean ± SD). Blue bar indicates the Km range of HIV-1 RT [8, 31]. T: untreated T cells, T + clof: T cells + 300 nM clofarabine, M: untreated macrophages, M + clof: macrophages + 300 nM clofarabine. d Clofarabine effect on HIV-1 proviral DNA synthesis. MAGI cells were incubated with DMSO (NI, HI, ND), 200 nM AZT, 300 nM clofarabine (Clof), or 50 nM raltegravir (Ralt) for 2 h prior to infection with pseudotyped HIV-1. Infection (black bars) was determined at 48 h post-infection, and reverse transcription (grey bars) efficiency was determined at 18 h post-infection. Data represent mean ± SD from three independent experiments and are expressed as percentage of vehicle control (DMSO, 100 %). NI no infection, HI heat inactivated virus, ND no drug