FIG 3.

Recruitment of neutrophils to peripheral tissues in the absence of CCR2 and localization of these cells in pyogranulomas with IMs. Mice were left uninfected (UI) or infected, and tissues were collected and processed as described in the legend to Fig. 2. (A to E) Cells from the indicated tissues were analyzed by flow cytometry. (A) Representative dot plots of CD19-CD3 (left) signals of viable splenocytes and Ly6G-CD11b (right) signals of gated CD19⁻ CD3⁻ cells of C57BL/6 (top) or CCR2^−/− (bottom) mice at 7 dpi. (B to E) The numbers of neutrophils from BM (B), spleen (C), MLN (D), and liver (E) of uninfected mice or mice at the indicated days postinfection were determined and are plotted. Means and standard errors shown are the summaries of data from two or more independent experiments. Significant differences between C57BL/6 and CCR2^−/− mice were calculated with a Mann-Whitney test, P values of <0.05 are indicated. In panel D, the numbers of polymorphonuclear leukocytes in MLNs of uninfected C57BL/6 and CCR2^−/− mice were 2,166 ± 410 and 7,943 ± 1,755, respectively. (F and G) Representative confocal microscopic images of individual and overlaid immunofluorescent signals, as indicated, from frozen sections of MLNs of CCR2-GFP mice infected with 32777 at 3 dpi (F) and 6 dpi (G).