Extended Data Fig. 1: FcγR expression profile of engineered cell lines and assessment of in vitro ADE activity.
From: Human FcγRIIIa activation on splenic macrophages drives dengue pathogenesis in mice
(a) FcγR expression on K562 was assessed by flow cytometry using antibodies against FcγRI, FcγRIIa, FcγRIIb, and FcγRIII (shaded histogram). Corresponding isotype control is shown in open histograms. (b,c) The in vitro ADE activity of the anti-DENV mAb C10 expressed as afucosylated (afuc) human IgG1 was assessed in U937 (black) vs U937-panFcγR (purple) cells. Percent (%) infection was assessed by flow cytometry. Area under curve (AUC) was calculated for each cell type and compared by two-tailed unpaired t-test. Figure shows one representative experiment (mean ± s.e.m) out of three performed in duplicates and normalized AUC from the three independent experiments. (d) The in vitro ADE activity of fucosylated (fuc) human IgG1, afucosylated (afuc) human IgG1, or Fc null variant was determined by multi-step viral yield on 6-, 15-, 24-, and 32-hours post-infection. Percent (%) infection was assessed by flow cytometry and compared between fucosylated and afucosylated hIgG1 by two-way ANOVA (Bonferroni post hoc analysis adjusted for multiple comparisons). Figure shows one representative experiment (mean ± s.e.m) performed in triplicates.
