A Novel CD48-Based Analysis of Sepsis-Induced Mouse Myeloid-Derived Suppressor Cell Compartments

Figure 3

Granulocytic and monocytic MDSC subsets identified by CD48 expression. (a) Bone marrow, spleen, and blood cells were collected from naïve and 7-day CLP Gfi1:GFP knock-in mice. MDSCs were gated as CD11b⁺Gr-1⁺ and sorted by CD48 expression. G-MDSCs were identified as CD11b⁺Gr-1⁺CD48⁻ and M-MDSCs were identified as CD11b⁺Gr-1⁺CD48⁺ cells. (b) Representative photomicrographs (images of bone marrow are shown) of Wright–Giemsa stained MDSC subsets, Bar = 10 μm. (c) Purity analysis of each MDSC subset by cell morphology. (d) Gfi1:GFP expression in CD11b⁺Gr-1⁺CD48⁻ and CD11b⁺Gr-1⁺CD48⁺ cells. Representative flow cytometry data are shown. (e) Purity analysis of MDSC subsets by Gfi1 expression. (f) Gfi1:GFP knock-in mice were treated with LPS (i.p. 10 mg/kg) or by CLP surgery. Bone marrow, splenic, and blood cells from 3 h LPS treatment and days 1, 7, and 12 CLP surgery were assayed. The purity of CD11b⁺Gr-1⁺CD48⁻ M-MDSCs was evaluated by Gfi1 expression. Representative graphs of bone marrow flow cytometry assay are shown. (g) Statistical analysis of flow cytometry data. Data are mean ± SEM of 5–8 mice per group.

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