Skip to main content
. 2023 Nov 2;7(1):13–27. doi: 10.1093/abt/tbad025

Figure 2.

Figure 2

Prevention of Fc-FcγR crosslinking by Fc engineering decreases LILRB4 internalization in FcγR high monocytic AML. (A, B) The expression level of FcγRs on THP-1 (A) and Mono-mac-6 (B) cells were checked by flow cytometry. All cells except debris were gated on an FSC-A/SSC-A plot. DAPI− cells were then gated and analyzed. Mean fluorescence intensity of FcγRs on THP-1 and Mono-mac-6 was measured by flow cytometry (n = 2). (C) Representative graphic of h128-3/N297A Fc-mutant IgG1 antibody with significantly decreased affinity for FcγRs. (D) Binding of WT h128-3 or h128-3/N297A to LILRB4 as assessed by ELISA (n = 2). (E, F) LILRB4 internalization induced by h128-3, h128-3/N297A or hIgG isotype control in THP-1 (E) or Mono-mac-6 (F) cells (n = 4). In brief, 2.5 × 105 cells were treated with 1 μg/ml of antibodies at 37°C for 24 h before measurement of LILRB4 internalization by flow cytometry.