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. 2016 Jun;433:51–58. doi: 10.1016/j.jim.2016.03.002

Fig. 3.

Fig. 3

ADCC assay and mAbs surface staining conducted with cells coated with HIV-1 Bal gp120. A. Comparison of RFADCC assay layout with CEM NKr CCR5 vs. EGFP-CEM-NKr-CCR5-SNAP cells. CEM NKr CCR5 were stained with PKH-26 and CSFE (Gomez-Roman et al., 2006), while EGFP-CEM-NKr-CCR5-SNAP cells were stained with Alexa Fluor 647-SNAP tag dye. Killing by C11 mAb (1 μg/ml) is determined as loss of CSFE in PKH26-positive CEM NKr CCR5 cells or loss of GFP in SNAP-positive EGFP-CEM-NKr-CCR5-SNAP cells. B. Cytotoxicity curves for gp120-coated EGFP-CEM-NKr-CCR5-SNAP. The ADCC results are representative of three independent assays and the bars indicate the range of the values of cytotoxicity of duplicate samples. The binding of 1 μg/ml Alexa-Fluor 647-conjugated mAbs C11 (C and F), N5-i5 (D and G) or N12-i2 (E and H) was compared in gp120-coated-CEM NKr CCR5 vs. EGFP-CEM-NKr-CCR5-SNAP cells.