Abstract
The adaptive immune system exhibits exquisite specificity and memory and is involved in virtually every process in the human body. Redirecting adaptive immune cells, in particular T cells, to desired targets has the potential to lead to the creation of powerful cell-based therapies for a wide range of maladies. While conventional effector T cells (Teff) would be targeted towards cells to be eliminated, such as cancer cells, immunosuppressive regulatory T cells (Treg) would be directed towards tissues to be protected, such as transplanted organs. Chimeric antigen receptors (CARs) are designer molecules comprising an extracellular recognition domain and an intracellular signaling domain that drives full T cell activation directly downstream of target binding. Here, we describe procedures to generate and evaluate human CAR CD4+ helper T cells, CD8+ cytotoxic T cells, and CD4+FOXP3+ regulatory T cells.
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Acknowledgements
This work was funded by the Human Islet Research Network (HIRN) Emerging Leader in Type 1 Diabetes grant U24DK104162-07 (LMRF), American Cancer Society (ACS) Institutional Research Grant IRG-19-137-20 (LMRF), South Carolina Clinical and Translational Research (SCTR) Pilot Project Discovery Grant 1TL1TR001451-01 (LMRF), Diabetes Research Connection (DRC) Grant IPF 22-1224 (LMRF), and Cellular, Biochemical and Molecular Sciences training grant T32GM132055 (RWC). Supported in part by the Flow Cytometry and Cell Sorting Shared Resource, Hollings Cancer Center, Medical University of South Carolina (P30 CA138313). Figures 1, 4, 6, 8, 9, and 12 were created with BioRender.com. Analyses in Figs. 7, 8, 10, and 11 were performed using GraphPad Prism for Mac. The flow cytometry results in Figs. 2, 4, 5, 6, 10, 12, and 13 were analyzed using FlowJo v10.8 Software (BD Life Sciences).
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Zimmerman, C.M., Robino, R.A., Cochrane, R.W., Dominguez, M.D., Ferreira, L.M.R. (2024). Redirecting Human Conventional and Regulatory T Cells Using Chimeric Antigen Receptors. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_15
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DOI: https://doi.org/10.1007/978-1-0716-3593-3_15
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