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. 2013 Jun 6;9(6):e1003406. doi: 10.1371/journal.ppat.1003406

Figure 7. Extramedullary myelopoiesis in malaria is established by mobilizing CCR2+ myeloid-restricted BM progenitors.

Figure 7

A. Expression of message for Ifng and IFN-γ induced chemokines in organ samples from pre-immune and mice infected with P. chabaudi at day 7. Three to four individual organ samples each were obtained from two independent experiments and analyzed by RT-PCR. B. Phenotype of LIN− cells in the spleen of C57BL/6 and Ccr2-null mice at day 7 after infection with P. chabaudi stained for indicated marker. Results in B.–E. are obtained form three infection experiments with 4–5 mice per group each. C. Absolute numbers of LIN− c-Kithi CD27+ CD16/32− (CD27+ CMPs) and CD16/32+ (GMPs) cells in the spleen of C57BL/6 and Ccr2-null animals in steady state or at day 7 after infection with P. chabaudi as quantified by flow cytometry. Results are shown as mean ± SEM from three independent experiments with 4–5 animals per group (**: P≤0.01, Mann-Whitney U-test). D. Changes in the absolute number of clonogenic myeloid progenitors (CFU-GM) in the BM and the spleen in uninfected mice and at day 7 after infection with P. chabaudi. Data for Ccr2-null mice and controls were obtained in three experiments with 4–5 samples per experiment and shown as mean ± SEM of total number of CFU-GM per organ (*: P≤0.05, **: P≤0.01, two-tailed Student's t-test). E. Phenotype of intrasplenic myeloid precursors during acute infection with P. chabaudi. Lineage-negative (Ter-119, CD3, CD11c, CD19, NK1.1) cells were stained for CD11b versus CD115 (M-CSF receptor) allowing the resolution of early myeloid precursors as c-Kithi Ly-6C−. F. Absolute numbers of early myeloid precursors (LIN− c-Kithi CD115+ CD11b− Ly-6C−) in the spleen of uninfected mice and at day 7 after infection. Representative staining and splenic cellularity were obtained from three independent infection experiments with 3–5 animals per group. Data represent mean ± SEM of cell number per spleen (**: P≤0.01, Mann-Whitney U-test). G. Course of malaria infection in C57BL/6 and Ccr2-null mice inoculated with blood stages of the malaria parasite P. chabaudi. Parasitemia was counted on the indicated days. Results for four infection experiments with 4–5 animals per time point in each experiment are shown as mean ± SEM (*: P≤0.05, **: P≤0.01, Mann-Whitney U-test).