Fig. 1.

Phenotype of plasmacytoid DC in the small intestine of mice. (A) Cells of the IE and LP preparation of the small intestine were stained with antibodies specific for CD3, CD11c, B220, MHCII, and Ly6C. CD3⁻ cells were analyzed for the expression of B220 and Ly6C (Left). The expression of MHCII and CD11c is shown for Ly6C⁺ B220⁺ (blue box, Center) and Ly6C⁻ cells (red box, Right). (B) Expression pDC-marker. pDC (CD11c⁺, B220⁺, and Ly6C⁺) of the IE and LP preparation were stained for PDCA-1 or 120G8 (solid lines) or isotype controls (shaded area) as indicated. (C) Cytospins of flow sorted IE mDC and pDC were stained with H&E (pDC: CD3⁻CD11c⁺B220⁺Ly6C⁺; mDC: CD3⁻CD11c⁺B220⁻Ly6C⁻). (Scale bars: 10 μm.) Representative data from one of two experiments are shown. (D) Cryostat sections of the small intestinal villi were stained for nuclei (DAPI, white), B220 (blue), CD3 (green), and 120G8 (red). The yellow bar in the upper left micrograph indicates how the positioning of pDC (120G8⁺B220⁺) relative to the epithelial layer was determined. (Top Right) Distance distribution of 150 pDC analyzed relative to the epithelium. (Middle and Bottom) Examples of the positioning of individual pDC with distances as indicated. (Scale bars: 10 μm.) (E) Flow cytometric analysis of pDC of the IE and LP preparation. Cells were stained with antibodies as indicated (solid lines) or isotype controls (shaded area) and gated on pDC (CD11c⁺, B220⁺, and Ly6C⁺). Shown are representative data from four independent experiments with cells pooled from two to six mice each.