Figure 7.

(A) DART-molecules with CD16-targeting arms bind to the surface of human NK cells (from n = 4 donors) by flow cytometry analysis. Data are reported as mean fluorescent intensities (MFI), error bars represent standard deviation. (B) Redirection of cord blood mononuclear cell (CBMC)-derived NK cells (from n = 12 CBMC samples, blue circles) for elimination of HIV-1 4403bmC5-infected autologous CD4⁺ T cells by 7B2 × CD16 or control (4420 × CD16) DART molecules in the absence of IL-15 (filled symbols) or after overnight incubation with 10 ng/mL IL-15 (open symbols). Results of assays performed with adult NK cells from three PBMC samples, in absence of IL-15, are included for comparison (black filled squares). Data are positive area under the dilution curve (pAUC) from 24 h killing assays with a NK cell to CD4⁺ T cell ratio of 5:1. (C) Similar levels of HIV x CD16 DART molecule-redirected killing of autologous HIV-1 4403bmC5-infeced CD4⁺ T cells were observed in assays performed with A32 × CD16 DART molecules and A32 × CD16 DART molecules combined with 7B2 × CD16 DART molecules (n = 3 CBMC-derived, IL-15 treated, NK cell samples, assayed in duplicate). (D) Redirection of CBMC (n = 2 samples, assayed in duplicate) for elimination of HIV-1 4403bmC5-infected autologous CD4⁺ T cells by 7B2 x CD3 DART molecules, 7B2 × CD16 DART molecules, or 7B2 x CD3 and 7B2 x CD16 DART molecules used in combination. Twenty four hours killing assay with an IL-15 treated whole CBMC to CD4⁺ T cell ratio of 30:1. Data in (C,D) represent mean +/– standard deviation.