Fig. 3: Ova-IC generated nAPCs retain neutrophil functions, promote naive CD4+ and CD8+ T cell proliferation and generate immunogenic cytokines. | Nature Communications

Fig. 3: Ova-IC generated nAPCs retain neutrophil functions, promote naive CD4+ and CD8+ T cell proliferation and generate immunogenic cytokines.

From: FcγR engagement reprograms neutrophils into antigen cross-presenting cells that elicit acquired anti-tumor immunity

Fig. 3

a Freshly isolated FcgRIIIB(3B)/γ−/− BMNs (Neut) or the same treated with anti-Ova, Ova-IC or SLE-IC and cultured to generate nAPCs were incubated with inactivated FITC-E. coli or IgG-coated, FITC-labeled latex beads and analyzed for FITC-uptake by flow cytometry. b Reactive oxygen species (expressed as relative light units/sec, RLU/s) generated over time by GM-CSF-primed 3B/γ−/− Neut and Ova-IC- or SLE-IC- generated nAPCs incubated with serum opsonized E. coli or zymosan. c, d Proliferation of CellTrace Violet-labeled CD4+ (OT-II) (c) and CD8+ (OT-I) (d) T cells after co-culture with Ova, Ova-IC or vehicle (−, GM-CSF alone) generated nAPCs of indicated genotypes assessed by CellTrace Violet dilution. In d, vehicle generated nAPCs pulsed with Ova SIINFEKL-peptide (pSIINF) is a positive control. Representative profiles for CD8+ T cells (d) are shown. e, f CellTrace Violet-labeled CD4+ (e) or CD8+ (f) T cells co-cultured with Ova- or Ova-IC generated nAPC, and Ova or Ova-IC treated splenic monocyte-derived (mDC) or Flt3L-induced splenic DCs, and analyzed as in (c, d). g, h Cytokine concentrations in supernatant of Ova- and Ova-IC generated nAPCs (g) and splenic cDCs (h). Data are mean ± s.e.m. For c–f one-way analysis of variance and Dunnett’s multiple comparison test; g, h Multiple t test between pairs of samples. *p < 0.05, **p < 0.005.

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