Fig. 3. Distinct dynamic behaviors from different neutrophil subtypes.

10⁶ GFP⁺ neutrophils isolated from UBC-GFP mice were intravenously (i.v.) injected in C57BL6/J mice that were i.p. infected with 10⁷ PFU of NYVAC-C Δ3 virus and the spleen was imaged from 4 to 5 h post infection (a). Multiphoton intravital microscopy dot plot determined by intensity and volume mean of GFP⁺ neutrophils from spleen of NYVAC-C Δ3-infected mice (b). Cell track plot of Nα and Nβ neutrophil from spleen of NYVAC-C Δ3 infected mice from 4 to 5 h post infection (c). Violin plots of displacement length (μm) (d) and speed (μm/min) (e) of Nα and Nβ neutrophils from 4 to 5 h post infection. Dot plots based on directionality and arrest coefficient of neutrophil subset activities (arrested: black; patrolling: red) (f). Percentages of arrested and patrolling Nα and Nβ from 4 to 5 h post infection (g). Nα (gray), Nβ (green) neutrophil and cognate CD8 T cells (cyan) from spleen of NYVAC-C Δ3-infected mice from 4 to 5 h post infection (h). Violin plots of percentage of time in contact of Nα and Nβ (i), graph bars of percentage of Nα and Nβ in contact with CD8 T cells (j), and violin plots of differential distance of Nα and Nβ to closest T cell (k). Graphs show mean ± SEM; each point represents an individual mouse. Data are representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.