Extended Data Fig. 4: Characterization of B7H3 and PDL1 expression across tumor and immune compartments.
From: A trispecific antibody engaging T cells with tumour and myeloid cells augments antitumour immunity
(a) PBMCs were co-cultured with SKOV3 tumor cells in the presence or absence of B7H3xCD3xPDL1 (Tri-TCE). Flow cytometry was performed before and after treatment to assess the surface expression of B7H3 and PDL1 on SKOV3 tumor cells (left panels) and CD11b+ myeloid cells (right panels). Representative histograms from one of three independent experiments are shown. (b) SKOV3 cells were treated with or without IFNγ (50 ng/ml) for 24 h. PDL1 surface expression was then assessed using flow cytometry. (c-d) Patient-derived tumor suspensions were treated with control IgG or Tri-TCE (B7H3xCD3xPDL1) in PDTS samples. (c) Representative flow cytometry plots showing gating strategy, Histograms show surface expression of B7H3 and PDL1 on tumor cells (CD45-) and CD11b+ myeloid cells before and after treatment. (d) Quantitative results (n = 3 independent experiments, mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test). (e) In the patient-derived tumor suspension (PDTS) model, tumor and immune compartments were distinguished by CD45 expression, and immune subsets were further gated into CD11b+ myeloid cells and CD3+ T cells. Flow cytometry was performed to assess B7H3 surface expression in each subset. (f) Quantification of B7H3+ cells among tumor and myeloid compartments in PDTS samples (n = 33 patient tumors, mean ± SD, paired two-tailed Student’s t-test).
