Improving the gene transduction efficiencyof chimeric antigen receptor (CAR) in Jurkat cellsthrough spinoculation

Background: Chimeric antigen receptor T cell (CAR-T) therapyis a type of immunotherapy in which T cells are geneticallyengineered to more efficiently target and eliminate cancer cells.In this method, it is critical to produce an adequate number ofengineered T cells. Considering that T cells are hard-to-transducecells, increasing the rate of viral vector transduction has asignificant effect on the success rate of this treatment method.Materials and Methods: Different transduction methods, includingspinoculation, polybrene, and spinoculation plus polybrenewere used to transduce Jurkat cells in order to determine the most effective transduction method. In this study, the concentratedviral stocks were transduced in untransduced and pretransducedJurkat cell lines to produce engineered CAR-T cells.Results: Flow cytometry analysis showed that spinoculationplus polybrene in concentrated virus (MOI=۱۰) had the highesttransduction rate of Jurkat cells (about ۸۱%). The transductionrate in untransduced Jurkat cells using concentratedvirus (MOI=۵) and spinoculation plus polybrene was ۷۵%,whereas for pretransduced Jurkat cells and spinoculation pluspolybrene and spinoculation were ۵۸% and ۴۷%, respectively.Transduction of untransduced Jurkat cells with concentratedvirus(MOI=۵) and polybrene without the spinoculationin hadthe lowest transduction’s rate of ۶%.Conclusion: The results indicated that the viral transductionrate using spinoculation plus polyberene was more efficient forJurkat cells compared to other groups. The production of CARTcells can be enhanced using this method which improves thevirus transduction conditions in manufacturing process.

کلیدواژه ها:

Chimeric antigen receptor ، Immunotherapy ، Spinoculation ، Polybrene ، Jurkat cell