Fig. 5.

ADCC assay using cells infected with HIV-1 BaL IMC. Left panel. EGFP-CEM-NKr-CCR5-SNAP cells were spinoculated with HIV-1 Bal molecular clone at 2000 rpm for 2 h at 12 °C. After 5 days of co-culture with the virus, cells were washed twice, labeled with Alexa Fluor 647-SNAP tag dye and incubated with dilutions of mAbs (C11, N5-i5, N10-U1 or Synagis) for 15 min at RT, then PBMC were added to the reaction for 3 h at 37 °C. At the end of the incubation, the samples were washed with PBS, fixed with 1% paraformaldehyde and analyzed by flow cytometry. The ADCC data represent the typical results obtained in three independent experiments. Upper right panel. The efficiency of the infection was evaluated by staining of the cells with live/dead (not shown), CD4-APC and p24-PE. Lower right panel. Binding of infected EGFP-CEM-NKr-CCR5-SNAP cells with 5 μg/ml Alexa Fluor-647-conjugated mAbs C11 (panel A), N5-i5 (panel B) or N10U1 (panel C).