Comparison of the EURT assay with the gold standard in HIV entry, the Blam-Vpr assay.

<p>To compare the EURT assay with the widely used Blam-Vpr assay, virions were produced by transfection of HEK293T cells using either reporters. A) Viruses were produced with an R5- or an X4- tropic Env (JR-FL and NL4-3, respectively), then used to challenge SupT1 and monocyte-derived dendritic cells (MDDCs, derived upon incubation of blood monocytes with GM-CSF and IL4 for 4 days). Envelope-receptor specificity was then measured according to the two viral entry assays in lymphocytes (SupT1) and in myeloid cells (MDDCs) either after cell lysis and luciferase activity measurement, or following the incubation of target cells with the fluorescent dye CCF2 and flow cytometry. B) SupT1 were used as target cells to compare the inhibitory rates that could be retrieved upon usage of Vpr-Blam- or EU-repRNA bearing HIV-1 X4 particles in the presence of the HIV-1 fusion inhibitor T20. The graphs present averages and SEM obtained with 3 independent experiments. * p≤0.05 after a Student t test between WT and the respective indicated conditions.