Figure 1: Distribution and phenotype of slanDCs in human tonsils.

Sections are from representative FFPE human tonsils and are stained as indicated by labels (CKP, pan-cytokeratin; LYS, lysozyme). (a) DD2⁺ slanDCs surround the CKP⁺ epithelium of tonsil crypts and mostly appear as large and stellate cells (b) (n=10). (c) Flow cytometry analysis was performed on live CD45⁺/HLA-DR⁺-singlet cells from CD3⁺/CD19⁺- depleted tonsil cell suspensions. The panel shows the gating strategy used to identify CD141⁺DCs (blue gate), CD1c⁺DCs (orange gate), CD14^high MΦ (grey gate) and slanDCs (purple gate), within the HLA-DR⁺/CD11c⁺ cells. By the same strategy, pDCs were identified within the HLA-DR⁺/CD11c⁻ cells (green gate). A representative experiment out of three ones performed with similar results is shown. (d) Bar graph shows the percentage of each DC subset and macrophages within the HLA-DR⁺ cells (mean±s.d.; n=5). (e) The histogram shows the relative expression of M-DC8 by each DC and MΦ populations. Data are representative of similar results obtained from five tonsil preparations. (f–m) Panels show representative DD2⁺ slanDCs co-stained with antibodies towards a set of markers (n=10), as labeled. Sections are counterstained with Meyer’s haematoxylin. Original magnifications: 40X (a, scale bar, 500 μm); 400X (b,f–m, scale bar, 50 μm).