Mice per group. Outcomes are representative of no less than two independent experiments with three to four mice per group (n = 6 animals per genotype). (F) IF staining of granulomas for the mannose receptor (red) and DAPI (blue). White arrowheads depict mannose receptor+ cells. (G) Flow cytometric quantification of mannose receptor+ cells from dissociated lung tissue. (H) Lung Arg1 and ChiA expression in naive and Sm egg-challenged WT and Retnla/ mice. , P 0.001; , P 0.05. (I) Masson’s trichromestained granulomas from WT and Retnla/ mice. White arrowheads, collagen stain. Bars, 50 . Results (imply SEM of three to four mice) are representative of two to 3 independent experiments (n = 60 per group).JEM VOL. 206, April 13, 2009activated CXCR4 drug macrophages, lymphoid cells, granulocytes, and multinucleated giant cells (Fig. 3 D, left). In contrast, Sm egg-challenged Retnla/ mice exhibited far more extreme inflammation surrounding the egg (Fig. 3 D, proper), which integrated a considerable increase inside the imply region of inflammation surrounding the granuloma (Fig. 3 E). Collectively these data indicate that RELM- deficiency final results in exacerbated Sm egg-induced pulmonary inflammation.AAMac responses are enhanced in Sm egg-challenged Retnla/ mice Provided that RELM- can be a signature gene of AAMacs, we hypothesized that RELM- deficiency might affect expression of other AAMac-derived genes or the CCR9 web recruitment or function of AAMacs. IF staining of mannose receptor+ cells in the granulomas from Sm egg-challenged WT and Retnla/ mice indicated that there was no impairment in the recruitment of mannose receptor+ AAMacs in the absence of RELM- (Fig. three F, red), and quantification of the mannose receptor+ cells by flow cytometric analysis of dissociated lung tissue revealed equivalent frequencies of mannose receptor+ AAMacs within the Sm egg-challenged WT and Retnla/ mice (Fig. 3 G). To examine AAMac responses after Sm egg challenge, lungs from naive and Sm eggchallenged WT and Retnla/ mice had been analyzed for expression of the AAMac genes Arg1 (Arginase 1) and ChiA (acidic mammalian chitinase) by real-time PCR. In WT mice, Sm egg challenge resulted in a 17-fold induction of Arg1 over naive controls (Fig. three H). In contrast, there was a 70-fold induction of Arg1 in Sm egg-challenged Retnla/ mice. Furthermore, despite the fact that Sm egg challenge of WT mice resulted inside a fourfold induction of ChiA, we observed a ninefold induction of ChiA in Retnla/ mice (Fig. 3 H). The increased recruitment of mannose receptor+ cells into the granulomas, coupled with substantial increases in levels of Arg1 and ChiA mRNA in Retnla/ mice, indicates that the AAMac responses are elevated in the absence of RELM-. Offered that a single proposed function of AAMacs would be to promote fibrosis (19, 37), in aspect by way of mediating collagen synthesis, we tested the hypothesis that Retnla/ mice may possibly exhibit variations in collagen deposition inside the Sm egg-induced granulomas. Consistent with elevated AAMac responses, Masson’s trichrome staining of your lung sections revealed that Retnla/ mice exhibited elevated collagen deposition within the egg-induced granuloma in comparison with WT mice (Fig. three I, arrowheads). Together, these data demonstrate that Sm egg-induced AAMac responses were increased in the absence of RELM-.draining mediastinal LNs (Fig. 4 A), comparable frequencies of lymphocytes inside the BAL and lung tissue (Fig. S3, A and B), and an equivalent boost inside the frequency of proliferating LN CD4+ T cells (Fig. four B) at day eight just after chall.
