Upregulation of Retinal Dehydrogenase 2 in Alternatively Activated Macrophages during Retinoid-dependent Type-2 Immunity to Helminth Infection in Mice
Figure 5
Type-2 inflammatory cells express Raldh2 and Raldh3, with Raldh2 most highly expressed in macrophages.
(A) qRT-PCR analysis of retinal dehydrogenase (Raldh) expression (isoforms 1–3) in hepatic inflammatory infiltrates. Expression is normalized to HPRT and presented as fold-change above the average expression in LCMV samples. n = 3–5 mice per group. (B) Fluorescence microscopy of a hepatic granuloma co-stained with antibodies recognizing CD11b (green) and Raldh (red). Arrows point to cells within granulomas that co-stain for CD11b and Raldh. Scale bar = 50 µm. (C) qRT-PCR analysis of Raldh isoform expression in sorted macrophages (Mac), eosinophils (Eos), and T cells. Expression levels were normalized and presented relative to HPRT expression. Expression levels of Raldh isoforms are presented here on the same scale (different from panel A) in order to illustrate that Raldh2 is the most highly expressed isoform. Hepatic leukocytes from S. mansoni-infected mice were pre-sorted by microbead selection of Siglec-F+ cells. Cell fractions were then sorted to >90% purity by FACS, according to the gating strategy shown. A modified Giemsa stain demonstrated the expected morphology of sorted cells. n = 3 mice per group. Error bars illustrate SEM; #p<0.001. Results are representative of two (C) or three (A, B) independent experiments.