Figure 10: Migratory properties by slanDCs from CCR patients and chemokine expression in M-TDLN.

(a,b) Freshly isolated PBMC from healthy donors and CCR patients were analysed for C5aR, CXCR4 and CX3CR1 expression on gated M-DC8⁺ slanDCs, by flow cytometry. (a) Data for each chemokine receptor are reported as mean fluorescence intensity (after the subtraction of the mean fluorescence intensity given by the correspondent isotype control antibody, mean±s.d.; n=6). (b) Representative histograms of CXCR4, CX3CR1 and C5aR expression by slanDCs from healthy donors and CCR patients are shown. (c) Graph shows the percentage of migrating slanDCs towards CXCL12, CX3CL1 and C5a in chemotaxis assays (n=4). (d–i) Sections from slanDCs positive M-TDLN representative cases (n=10) stained as indicated. CXCL12/SDF-1 and CX3CL1/Fractalkine are detected in tumour cells, nodal stromal cells and, for CXCL12/SDF-1, also in HEV endothelium (serial sections d,e and g,h). C5a is mainly expressed by nodal stromal cells (f,i), some of them expressing CD11c (blue, insert in f). T, tumour cells. Sections are counterstained with Meyer’s haematoxylin. Original magnifications: 100X (d,e, scale bar, 200 μm); 200X (f–h, scale bar, 100 μm) and 400X (i, scale bar, 50 μm).