An Indigenous Workflow for Generating CD19 CAR-T Cells

FDA-approved CD19 CAR T cell therapy for treating B-cell lineage malignancies gave impetus to the adoptive immune/cell and gene therapy field. Although tested to a considerable extent, side effects such as immune effector cell-associated neurotoxicity syndrome (ICANS), immune effector cell-associated hematotoxicity (ICAHTs), the high cost of the therapy, and often complicated logistics make the accessibility of this CAR therapy far from the reach of many eligible patients. Therefore, the development of a safe, cost-effective, streamlined indigenous workflow for T-cell expansion, transduction, and CAR T-cell characterization is essential in a preclinical as well as clinical setting. We have optimized a method to develop CD19 CAR T cells from peripheral blood T cells from healthy donors. In this chapter, we describe an indigenous protocol for the generation and characterization of CD19 CAR T cells via lentiviral transduction to understand the immunology or possible downstream applications.