Fig. 3
From: TNF-α x FcαRI bi-specific antibody potentiates neutrophil-mediated anti-tumor effects
Combination of TNF-α and FcαRI BsAb potentiates neutrophil A431 tumor killing and degranulation in vitro. (A) Representative dot plots of neutrophils after 4 h of co-culture with DiO-labeled A431 tumor cells in the presence of FcαRI BsAb, TNF-α or the combination. (B) Quantified trogocytosis of DiO-labeled A431 tumor cells by neutrophils in the experiments of Fig. 3A. Measured by flow cytometry and displayed as the percentage DiO+ neutrophils. (C) Neutrophil-mediated A431 tumor killing following 4 h of co-culture in the presence of FcαRI BsAb, TNF-α or the combination. Measured by CellTiter Blue assay and standardized against untreated controls. (D,E) Surface expression of neutrophil markers CD11b (D) and CD66b (E) following trogocytosis assays of Fig. 3A. Measured by flow cytometry and displayed as the geometric mean fluorescent intensity. (F) Secretion of lactoferrin in supernatants of trogocytosis assays of 2 A. Measured with ELISA. Graphs show means of N ≥ 3 experiments. Error bars indicate standard error. *P < 0.05; ****P < 0.00005 by 1-way ANOVA with Tukey’s multiple-comparison correction.
