Figure 4.

CB-pDCs show key functional features resembling primary pDCs. Unprimed and primed sorted CB-pDCs as well as primary pDCs were activated by TLR9 or TLR7 agonists for 24 hrs. (A) Quantification of pan-IFN-ɑ release in supernatant. (B) Representative flow cytometry dot plots showing expression of intracellular IFN-ɑ2 after 6 hrs (from CD45⁺CD123⁺CD303⁺) of 2 independent experiments. (C) Quantification of IFN-λ1, TNF-ɑ, CCL5, and IFN-β in supernatant. (D) Overlaid histograms comparing expression of activation markers on the surface of the different pDC types. (E) Quantification of marker expression on the surface of pDCs (from CD45⁺CD123⁺CD303⁺). (F) Scheme illustrating the set-up of pDC-NK cell co-culture assay. (G) Quantification of IFN-ɣ in supernatant upon co-culture of NK cells with CB-pDCs or pDCs. (H) Overlaid histograms showing activation marker expression on NK cells upon co-culture with CB-pDCs. (I) Quantification of activation marker expression on NK cells (from CD45⁺CD56⁺CD123^-) upon co-culture with pDCs. Results are shown as mean ± SEM of 2 independent experiments, (A, C, E) 6 independent CB donors and 4 independent primary pDC donors and (G, I) 5 independent CB donors, 4 independent pDC donors and 4 independent NK cell donors are depicted. *p<0.05; **p<0.01, ***p<0.001, ****p<0.0001, (A, C, E) two-way ANOVA or (G, I) one-way ANOVA with Tukey’s post-hoc test. See also Supplementary Figure S3 .