Author: Lippens, Carla; Duraes, Fernanda V.; Dubrot, Juan; Brighouse, Dale; Lacroix, Mathilde; Irla, Magali; Aubry-Lachainaye, Jean-Pierre; Reith, Walter; Mandl, Judith N.; Hugues, Stéphanie
Title: IDO-orchestrated crosstalk between pDCs and Tregs inhibits autoimmunity Document date: 2016_12_23
ID: sl8148ap_26
Snippet: WT BM-derived pDCs were generated in vitro for 7–8 days and purified using the Plasmacytoid Dendritic Cell isolation kit II (Miltenyi Biotec) according to manufacturer's instructions. Cell purity was assessed by flow cytometry using a Cyan™ ADP (Beckman Coulter) and exceeded 90%. pDCs were loaded or not with MOG35–55 peptide (10 μg/mL). 500 000 pDCs were seeded in 24 well plates with 300 000 2D2 CD4+ T cells or 300 000 pDCs were seeded in .....
Document: WT BM-derived pDCs were generated in vitro for 7–8 days and purified using the Plasmacytoid Dendritic Cell isolation kit II (Miltenyi Biotec) according to manufacturer's instructions. Cell purity was assessed by flow cytometry using a Cyan™ ADP (Beckman Coulter) and exceeded 90%. pDCs were loaded or not with MOG35–55 peptide (10 μg/mL). 500 000 pDCs were seeded in 24 well plates with 300 000 2D2 CD4+ T cells or 300 000 pDCs were seeded in 48 well plates with 100 000 2D2 CD4+ T cells or 100 000 2D2 CD4+ CD25− cells or 100 000 2D2 CD4+ CD25+ cells. Cells were co-cultured in complete RPMI medium for 16 h and pDCs were isolated again using the Plasmacytoid Dendritic Cell isolation kit II (Miltenyi Biotec) according to manufacturer's instructions.
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