Fig. 4: Induction of FcγRIII stimulation, NK activation, and ADCC by FD22. | Cell Discovery

Fig. 4: Induction of FcγRIII stimulation, NK activation, and ADCC by FD22.

From: A potent and broad CD4 binding site neutralizing antibody with strong ADCC activity from a Chinese HIV-1 elite neutralizer

Fig. 4

a Flow cytometry analysis of FD22 binding to the cell surface of H9 (IIIB) cells, TNF-α reactivated ACH-2 cells, and HL2/3 cells. VRC01 and the IgG isotype were used as controls. b FD22-mediated ADCC was evaluated using Jurkat-Lucia NFAT-CD16a reporter cells in the presence of H9 (IIIB) cells, TNF-α reactivated ACH-2 cells, or HL2/3 cells. FcγRIIIa signaling activity was quantified by measuring the luciferase signal from an NFAT-dependent reporter, with isotype control-induced signaling set to 1 for comparison. The ADCC V3-specific antibody 10-1074 served as a positive control. c Degranulation of NK cells upon FD22 engagement, as measured by CD107a surface expression. Intracellular staining for MIP-1β and IFN-γ release from NK cells in response to FD22. The data are representative of at least three independent experiments in which NK cells were isolated from the peripheral blood of healthy donors. d ADCC activity of FD22 against HIV-1-infected dells. The ADCC activity of FD22 was assessed using the LDH release cytotoxicity assay. H9 (IIIB) cells served as target cells, while PBMCs from healthy donors were used as effector cells at an E:T ratio of 6:1. FD22 was tested at concentrations ranging from 6.25 to 200 μg/mL.

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