Scheme 2.

Schematic representation of methodological approach. Human peripheral blood cells were isolated from whole blood through gradient centrifugation [28,29,31]. Primary CD4⁺ T cells were infected with a GFP-tagged HIV-1 NL4.3 virus for 5 days [30]. In method 1, cells were infected with HIV-1 and then treated with AZT-derivatives. In method 2, primary T cells were pre-treated with AZT-derivatives for 18 h, then washed to remove the drugs and allow for HIV-1 infection. To restore drug treatment, 10 µM of AZT-derivative was added post-infection. At the end of the 5 days, HIV-1 replication was assessed by comparing the frequency of HIV-1 infected cells in non-treated versus AZT-derivative treated conditions by flow cytometry.