FIG 3.

ADCC NK cell activation mediated by MAbs CH42 and CH43 incubated with NK-92-CD16 cells (which stably express GFP) and either HSV-1-infected SK-N-SH cells or HSV-1 gD-coated 96-well plates. (A) NK-92-CD16 cells (1 × 10⁵) were incubated with HSV-1-infected or uninfected SK-N-SH cells in the presence or absence of MAbs at 5 μg/ml (CH42, CH43, and A32 [a human IgG1 isotype control] or no antibody [No AB]) for 5 h. Cell surface CD107a staining was then performed as a marker of NK cell activation. The percentages of NK-92-CD16 cells (GFP positive) that expressed CD107a on their surfaces are shown on the y axis. (B) Dose-response curve of ADCC using serial dilutions of MAb CH42, CH43, or isotype control A32 with HSV-1-infected or uninfected SK-N-SH cells. (C) Dose-response curve of NK-92-CD16 cell activation using serial dilutions of MAb CH42, CH43, or isotype control A32 in HSV-1 gD-coated 96-well plates. The diluted antibodies were added to the gD-coated wells and incubated for 15 min, and 5 × 10⁵ NK-92-CD16 cells/well were added and incubated for 5 h. After washing with PBS, the cells were stained with 4 μg/ml APC-Cy7-conjugated anti-CD107a antibody and fixed with 10% paraformaldehyde. Activated degranulating NK cells (GFP⁺ CD107a⁺) were detected by flow cytometry. The percentages of CD107a⁺ NK-92-CD16 cells are shown. The error bars indicate standard deviations.