Matthew R Thakur, Emily M Flowers, Stephanie C Eisenbarth, Adam Williams
Storage of red blood cells (RBCs) increases the incidence of alloimmunization following transfusion. Mouse transfusion models have revealed the central role that splenic dendritic cells (DCs) play in storage-dependent RBC alloimmunization. The use of flow cytometry, immunofluorescence imaging, and T-cell proliferation assays combined with various genetically modified mouse strains has provided a detailed understanding of the processes by which DCs instruct the generation of alloantibodies. We and others have established a sequence of events in which transfused stored RBCs are taken up by splenic DCs, resulting in their activation and subsequent migration into T-cell zones. Once there, DCs drive MHCII-dependent T-cell activation and eventually alloantibody production. Herein, we described various methods that allow for the investigation of DC function in the context of alloimmunization.