Fig. 4: The effect of anti-stab1 antibody and FOXP3-ASO treatment in vitro.
From: Immunotherapy that improves response to chemotherapy in high-grade serous ovarian cancer
A Incucyte human macrophage phagocytosis curves (n = 3 donors). Green = anti-stab1 antibody treatment, dark-red = isotype antibody treatment, red = macrophages alone and cancer cells alone (as negative controls). B Area under the curve (AUC) of the curves in 4 A (n = 3 donors). Error bars at median and 95%CI. Two-side paired t-test. C Median florescent intensity (MFI) of TNFα, CD11C and PDL1 on macrophages using flow cytometry. Gating on single/live/CD14+ cells. Error bars at median and 95%CI. Two-way ANOVA test with Tukey correction. D Incucyte S3 curves showing T cell killing (n = 3 donors) of G164 ovarian cancer cell line. T cells cultured in anti-stab1 antibody (pink) or isotype-treated macrophage conditioned medium (blue) or no macrophage medium (green). Green and red curves = T cells or cancer cells alone as controls for naturally occurring cell death in both. Error bars at median and 95%CI. E AUC of the curves in 4D (n = 3 donors). One-way ANOVA with Tukey correction. F Flow cytometry analysis of T cells in 4D. Gating was done on live/singlet/CD4+ or CD8 + . One-way ANOVA with Tukey correction. G Time-course flow cytometry study of human T cells showing the effect of FOXP3-ASO over time (n = 3 donors). Gating was done on live/singlet/CD4+ or CD8 + . i) FOXP3 percentage of CD4, ii) MFI of Ki67 on CD4 + , iii) TBET percentage of CD4, iv) LAG3 on CD4, v) CTLA4 percentage on CD4, vi) MFI of Ki67 on CD8, vii) LAG3 percentage of CD8 cells and viii) Perforin+ cells in CD8. Error bars at median and 95%CI. Two-way ANONA t test with Tukey correction. H Incucyte S3 curves showing T cell killing of G164 cells plotted over time (n = 3 donors). Star=FOXP3-ASO treatment, circle= CTL-ASO treatment. Pink curves= T cells cultured without rhTGFβ. Purple curves= T cells cultured with rhTGFβ Error bars at median and 95%CI. I AUC of T cell killing curves in 4H (n = 3 donors). Error bars at median and 95%CI. Two-sided unpaired student t-test. rhTGFβ = recombinant human TGβ. CTL-ASO=control-ASO.
