Skip to main content
Figure 3 | Retrovirology

Figure 3

From: HIV-1 Nef promotes the localization of Gag to the cell membrane and facilitates viral cell-to-cell transfer

Figure 3

Nef enhances viral cell-to-cell transfer in primary CD4+ T cells. Primary CD4+ T cells were infected with VSV-G-pseudotyped WT- or ∆Nef in order to get similar levels of Gag (KC57) positive cells, or, as a negative control, left uninfected (NI). These cells were then co-cultivated with target lymphocytes pre-stained with carboxyfluorescein succinimidyl ester (CFSE) for 2h, and analyzed by flow cytometry. (a) Dot plot analysis of donors (upper panels) and targets (lower panels) in one representative experiment. The percentage of Gag (KC57) positive cells is indicated in the top right corner of the gated population. MFI is also indicated. (b) Percentages of Gag (KC57) positive primary CD4+ target cells in 5 independent experiments. (c) Contacts and virological synapses between infected donors (D) and uninfected CD4+ lymphocytes targets (T), visualized by immunofluorescence. Donor cells were co-cultivated with CFSE-labeled (green) target cells for 1h and stained for HIV-1 Gag proteins (red) using a polyclonal rabbit anti-Gag antiserum. A contact was defined as a tight interaction between the cells (upper panel). A virological synapse was defined as a cell conjugate in which a polarization of Gag proteins was visible at the contact zone (lower panel). (c, d, e). Quantification of the percentage of conjugates (d) and virological synapses (e) formed between donor and target cells. *p<0.05 (Mann Whitney test).

Back to article page