Fig 3. Disparate inflammatory monocyte responses at sites of primary and secondary infection.

Naïve wild type B6 mice (NaLm) or mice with a healed but persistent primary infection (LmLm) were challenged in the ear with 2x10⁵ L. major (A-F) or L. major-RFP (G-J). At the indicated times p.i. the ear dermis was analyzed as indicated. 0h time point data are prior to challenge. Statistical analysis refers to comparison between NaLm and LmLm groups (asterisks) or between time points (#). (A) The number of IFN-γ⁺CD4⁺ T cells/ear determined by multiplying the absolute number of CD3⁺CD4⁺ T cells per ear employing counting beads by the frequency of cells with the capacity to make IFN-γ following APC+L. major-Ag re-stimulation. (B) The frequency of cells with the capacity to make IFN-γ following APC+L. major-Ag re-stimulation. (C) The number of parasites per ear as determined by LDA. (D) The absolute number of CD11b⁺ cells per ear as determined employing counting beads. (E) The total number of the indicated populations per ear during primary (NaLm) or secondary (LmLm) infection. Cells were gated as generally described in S3 Fig. (F) Median fluorescence intensity of MHCII on the indicated populations over time. Data in (A-F) are from one experiment representing more than 3 similar experiments. Data are the Mean +/- error as indicated in the figure, n = 7–8 ears per time point. (G and H) Analysis of the frequency of the indicated populations within the infected RFP⁺CD11b⁺ (G) or total CD11b⁺ (H) population. (I) mRNA expression from the ears depicted in G and H, the values shown are the fold increase relative to expression in naive ears. Data are the mean +/- SEM. Data are pooled from two similar experiments. In (I) statistical analysis refers to comparison between NaLm and LmLm groups: ifnγ p<0.0001 on 10hr, d2, d4 and d8; cxcl10 p≤0.005 on 10hr, d2, d4, and d8; cxcl9 p≤0.0005 on 10hr, d2, d4, and d8; inos p<0.0001 on d2, d4, d8; ccl2 p = 0.0483 on d8; il10 *, p = 0.036, **, p = 0.004. Line refers to time points that were statistically significant. (J) Synchronous comparison of the ifnγ and inos expression kinetic taken from (I). ##, p≤0.0041 comparing inos 0hr with d2, 4, 8, and 14. ****, p≤0.0001 comparing ifnγ 0hr with 10hr, d2, 4, 8, and 14.