Microglia and macrophages differ in their inflammatory profile after permanent brain ischemia.

We studied the expression of pro- and anti-inflammatory molecules in microglia and infiltrating monocyte-derived macrophages after permanent Middle Cerebral Artery Occlusion (pMCAO). LysM-EGFP knock-in mice were used to distinguish between these two cell types, as peripheral myeloid cells are LysM-EGFP+, while microglia are not. This was confirmed with P2ry12 (a microglial specific marker), Iba-1 and EGFP immunostaining. The peak of LysM-EGFP+ myeloid cell infiltration was 72h post-ischemia, and were distributed evenly in the lesion core, surrounded by a dense region of microglia. Flow cytometry showed that a higher percentage of microglia expressed TNF-α at 3 (24.3% vs 1.4%) and 7 (18.8% vs 3.4%) days post-pMCAO as compared to infiltrating macrophages. Microglia and macrophages were purified by fluorescence activated cell sorting 72h post-ischemia to assess the mRNA expression of inflammatory markers. Macrophages upregulated expression of mRNA for arginase-1 (Arg-1) by 1000-fold, and IL-1β by 90-fold as compared to microglia. At the protein level, a significantly number of macrophages expressed Arg-1, while few if any microglia expressed Arg-1. However, IL-1β protein was not detected in macrophages by flow cytometry or immunofluorescence labeling of tissue sections. It was, however, detected in astrocytes along the lesion border. A PCR-array screen of 84 inflammatory genes revealed that pro-inflammatory chemokines and cytokines were predominantly upregulated in macrophages but down-regulated in microglia in the ischemic brain. Our results show clear differences in the inflammatory expression profiles between microglia and macrophages 72h post-ischemia which may shape repair and pro-regenerative mechanisms after stroke.