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. 2016 Jun 16;9:77–86. doi: 10.1016/j.ebiom.2016.06.008

Fig. 2.

Fig. 2

Stored RBCs activate splenic DCs. (a–c) Chimeric mice were generated with bone marrow from Zbtb46-DTR mice, as described in the Methods, and treated with diphtheria toxin prior to transfusion with HOD RBCs stored for 14 days. a) Representative flow cytometric analysis of CD11c+ MHC-II+ cDCs in spleens of mice treated with (+) or without (−) diphtheria toxin (DT). Dot plots were gated on live TCRβ− CD19− non-lymphocytes. Numbers on plots indicate the percentage of gated cells within the indicated gate. b, c) Anti-RBC antibodies in sera of mice treated with or without DT prior to stored HOD RBC transfusion, quantified by flow cytometric cross-match (b) or ELISA (c) as in Fig. 1. n = 6–10 mice/group; results are representative of 5 independent experiments. ****p < 0.0001; **p < 0.01. d–e) Expression of activation markers, CD86 and MHC-II, on spleen cDCs (gated as in a) from naïve mice (shaded) or mice injected i.v. 4 h prior (open histogram) with (d) LPS, stored HOD RBCs, stored GFP-expressing RBCs or (e) stored WT C57BL/6 RBCs. 1 representative mouse from 2 mice/group from 3 independent experiments.