Figure 1

Human vaginal mucosa contains four major subsets of myeloid-originated antigen-presenting cells (APCs). (a) Flow cytometry analysis of cells in human vaginal mucosa after enzymatic digestion. (Left panel) Live HLA-DR⁺ cells were gated and (middle panel) CD207⁺ cells (I) were gated. HLA-DR⁺CD207⁻ cells were further divided into four groups based on CD1c and CD14 expression (right panel: II, CD1c⁺CD14⁻; III, CD1c⁺CD14⁺; IV, CD1c⁻CD14⁺; V, CD1c⁻CD14⁻). Data are representative of 20 independent experiments. (b) Morphology of FACS-sorted subpopulations of vaginal cells (original magnification, × 100; bars=20 μm). Data are representative of four independent experiments. (c) FACS-sorted HLA-DR⁺CD207⁻CD1c⁻CD14⁻ cells were stained for CD34, CD54, CD66, pan-cytokeratin, or E/P selectin. (Upper panel) One representative data from three independent experiments using cells from different donors. (Lower panel) Percentage of positive cells has been calculated. Each dot represents data acquired from one donor. (d) Frozen tissue sections were stained for CD207 (green), CD14 (red), CD1c (light blue), and cell nuclei (dark blue) (original magnification, × 20; bar=100 μm). Data are representative of six independent experiments. (e, g) Flow cytometry analysis of the vaginal APC subsets. Tissues were digested with enzymes and cell suspension was stained with indicated antibodies and gated as in a. Gray histograms represent isotype controls. Data are representative of (e) 10 or (g) 6 independent experiments. (f) Frozen tissue sections were stained for (upper panel) CD1a (green), CD1c (light blue), and CD14 (red) and (lower panel) CD163, (green), CD1c (light blue), and CD14 (red) (original magnification, × 20; bar=100 μm). Data are representative of six independent experiments. All of the independent experiments were performed with tissues or cells from different donors. DC, dendritic cells; FACS, fluorescence-activated cell sorting; LC, Langerhans cells; LP, lamina propria.

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