Supplementary Figure S4 from Human Tumor–Associated Macrophages and Neutrophils Regulate Antitumor Antibody Efficacy through Lethal and Sublethal Trogocytosis

Supplementary Figure S4. (A) Cumulative results showing the kinetics of GFP+A431 tumor cell growth when co-cultured with PBN isolated from lung cancer patients at 50:1 E:T ratio in the presence of cetuximab (IgG) and IgA anti-EGFR Ab (IgA) using the IncuCyte® Live Cell Analysis System. One-way ANOVA with Turkey’s multiple comparisons test. Number of patients included in each analysis is indicated on the graphs. All data represented as mean ± SEM. (B) Cumulative results showing the kinetics of GFP+A431 tumor cell growth when co-cultured with blood and tumor FcR+ effectors for 48 hrs at 50:1 E:T ratio in the presence of IgA2 isotype control Abs in the IncuCyte® Live Cell Analysis System. One-way ANOVA with Dunnett's multiple comparisons test. All data represented as mean ± SEM. (C) Representative dotplots showing ADT mediated by TAM pretreated with cytohalasin D (1uM) or blocking anti-CD64 F(ab')2 and anti-CD32 F(ab')2 fragments (5ug/ml). TAM were co-cultured with PKH67+ A431 cells at a ratio 3:1 in the presence of cetuximab (1ug/ml) for 2 hrs. (D) Cumulative flow cytometry results showing the trogocytosis mediated by indicated blood and tumor myeloid effectors co-cultured with PKH67+A431 cells for 12 hrs at a 50:1 E:T ratio in the presence of IgG1 and IgA2 isotype control Abs (1ug/ml). Indicated FcR+ effectors were freshly isolated for all experiments.