Modeling the Application of Chimeric Antigen Receptor T Cell Therapy to Eliminate Alloantibodies in Mice.

Chimeric antigen receptor (CAR) T cell therapy has begun to show clinical promise as a strategy for immune-modulation in non-malignant conditions. In a murine model, we have demonstrated the utility of CAR T cells targeting B cells and plasma cells for the elimination of alloantibodies that cause allograft rejection. We describe a method for producing murine CAR T cells, their functional assessment in vitro, as well as their application in an in vivo transplant model. Our methods include a CAR production phase that involves retrovirus production, T cell isolation, activation, transduction and passage, and assessment of CAR expression by flow cytometry. Then, we describe methods for studying CAR T function in vitro using a cytotoxicity assay and in vivo using an islet transplant model.