Abstract
Background: Current advancements in the field of chimeric antigen receptor (CAR) therapy, particularly U.S. FDA approval of
Kymriah and Yescarta, heralds a new era of cancer treatment. This rapid progress in technology has urged more countries and
institutions to keep pace with the fast-growing and developing technology of producing CAR T cell-based therapies in the race
to develop new cancer-targeting drugs. Hence, for stepping in line with global advances and to pave the way for subsequent
preclinical and clinical studies, we have established a development protocol for a cancer-targeting CAR T cell; we have chosen
CD19 CAR T cell as a well-defined model to set-up T cell expansion, activation, and viral transduction as the prerequisites for
diverse CAR T cell therapies.
Methods: T cells from peripheral blood mononuclear cells (PBMCs) were activated and expanded. CD19 CAR lentiviral particles
were produced in the Lenti-X™ 293T Cell Line using PolyFect Transfection Reagent.
Results: Activation protocol resulted in (65 ± 4%; P = 0.046) increase in the rate of activated T cells 24 hours after the initiation
of the procedure. The expansion methodology resulted in a high purity of the T cell population (96 ± 3%) in the pool of PBMCs
within 14 days of the procedure. Finally, 35 ± 6% of T cells were transduced with CD19 lentivirus with MOI of 3.
Conclusion: Collectively, the results of this study prove that we have successfully overcome the first hurdle on the road to reach
CAR T cell technology which is the prerequisite for developing preclinical and clinical phases of CAR therapy in settings with
basic resources