Figure 1

Uptake of infected and uninfected neutrophils by DCs. (a) The eGFP^hi neutrophils recovered from the ear dermis 12 h after infection of C57BL/6 LYS-eGFP mice with 2 × 10⁶ Lm-RFP metacyclic promastigotes were sorted to obtain uninfected eGFP^hiRFP⁻ and infected eGFP^hiRFP⁺ neutrophils. Data are representative of several independent experiments. (b) Frequencies of annexinV⁺ 7-AAD⁻ uninfected eGFP^hiRFP⁻ or infected eGFP^hiRFP⁺ neutrophils recovered from the ear dermis at 2 and 16 h after infection with 2 × 10⁶ Lm-RFP metacyclic promastigotes. Values shown are the mean percentage±1 S.D., n=3–4 ears/group/time point. Data are representative of two independent experiments. (c) Representative dot plots of DCs cultured with eGFP^hiRFP⁻ and eGFP^hiRFP⁺ dermal neutrophils for 16 h (1 : 2 neutrophil/DC ratio) and analyzed for expression of CD11c and GFP signal. Gates represent populations of CD11c⁺GFP⁻ and CD11c⁺GFP⁺ cells. (d) Representative histogram plots of RFP expression by post-sorted populations of CD11c⁺GFP^- (black line) and CD11c⁺GFP⁺ (gray filled) cells recovered from co-culture of DCs with eGFP^hiRFP⁺ dermal neutrophils, gated as shown in (c). (e) Confocal microscopic images of sorted CD11c⁺GFP⁺ cells recovered from culture of DCs with eGFP^hiRFP⁻ and eGFP^hiRFP⁺ dermal neutrophils. DCs were derived from β-actin CFP mice and are shown in blue, eGFP^hi neutrophils are shown in green and Lm-RFP parasites are shown in red