Figure 3

Antigen presentation function of DCs that have captured infected and uninfected neutrophils. (a–c) Sorted CD11c⁺ (no exposure to neutrophils), CD11c⁺GFP⁻ and CD11c⁺GFP⁺ cells (following overnight culture with uninfected or infected neutrophils) were cultured with CFDA-labeled OT-I CD8⁺ T cells for 3 days in the presence of soluble OVA. (a) Representative histogram plots of CFDA fluorescence of CD8⁺CD3⁺ gated cells showing the proliferative response. Numbers represent the frequency of cells with reduced CFDA content. (b) CFDA-labeled OT-I CD8⁺ T cells with reduced CFDA content (mean percentage±1 S.E.M., calculated from the means of three independent experiments, two replicates per experiment). (c) IFN-γ levels in supernatants of the cultures described in (b) (mean concentration±1 S.E.M., calculated from the means of three independent experiments, two replicates per experiment). (d) Histogram plots of CFDA fluorescence of OT-I CD8⁺CD3⁺ gated cells showing the proliferative response following 3 days of co-culture with CD11c⁺, CD11c⁺GFP⁻ and CD11c⁺GFP⁺ cells in the presence of SIINFEKL peptide. Numbers represent the frequency of cells with reduced CFDA content. (e) Representative dot plots showing the gates used for sorting of the CD11c⁺ (no exposure to Lm-RFP) and CD11c⁺RFP⁺ cells following direct infection with Lm-RFP metacyclic promastigotes for 12 h (1 : 1 parasite/DC ratio). (f) Representative histogram plots of gated CFDA-labeled OT-I CD8⁺ T cells showing the percentage of the proliferative response after co-culture with CD11c⁺ and infected CD11c⁺RFP⁺ cells for 3 days in the presence of OVA antigen. (g) CFDA-labeled OT-I CD8⁺ T cells with reduced CFDA content (mean percentage± 1 S.D., calculated from three independent experiments)