C5aR+ lysozyme-expressing dendritic cells (LysoDCs) comprise a unique cell population in the Peyer's patch (PP) subepithelial dome (SED).

CD11chi cells can be subdivided into six different cell subsets:plasmacytoid DCs (pDCs), CD8a+ DCs, CD11b+ DCs, double-negative (DN) DCs, LysoDCs, and LysoMacs. Among these, LysoDC (CD4-) and LysoMacs (CD4+) are classified as CD11chiMHC class II+BST2intSIRPalphahi lysozyme+ cells, depending on whether they express CD4. Here, we determine that C5aR expressed in LysoDC and LysoMacs. We used microarrays to determine the lineage of C5aR+ LysoDC and C5aR- LysoDC. PP cells were prepared by collagenase D (Roche)/DNase I buffer and after removing dead cells using a dead cell removal kit, CD11c+ cells were enriched using anti-mouse CD11c microbeads. C5aR+ LysoDC, C5aR- LysoDC, and LysoMac were sorted using FACSAria III and total RNA was prepared using an RNeasy PLUS Micro kit.