Engineered Jurkat Cells for Targeting Prostate-Specific Membrane Antigen on Prostate Cancer Cells by Nanobody-Based Chimeric Antigen Receptor

Engineered Jurkat Cells for Targeting Prostate-Specific Membrane Antigen on Prostate Cancer Cells by Nanobody-Based Chimeric Antigen Receptor

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Background: Recently, modification of T cells with chimeric blunt wraps antigen receptor (CAR) has been an attractive approach for adoptive immunotherapy of cancers.Typically, CARs contain a single-chain variable domain fragment (scFv).Most often, scfvs are derived from a monoclonal antibody of murine origin and may be a trigger for host immune system that leads to the T-cell clearance.Nanobody is a specific antigen-binding fragment derived from camelid that has great homology to human VH and low immunogenic potential.Therefore, in this study, nanobody was employed instead of scFv in CAR construct.

Methods: In this study, a CAR was constructed based on a nanobody against PSMA (NBPII-CAR).At first, Jurkat cells were electroporated with NBPII-CAR, and then flow cytometry was performed for NBPII-CAR expression.For functional analysis, CAR T cells were co-cultured with prostate cancer cells and analyzed for IL-2 secretion, CD25 expression, and cell proliferation.Results: Flow cytometry results confirmed the expression of NBPII-CAR on the transfected Jurkat cells.Our data showed the specificity of engineered Jurkat cells against prostate cancer cells by not only increasing the IL-2 cytokine (about 370 pg/ml) but also expressing the T-cell activation marker CD25 (about 30%).

In addition, proliferation of engineered Jurkat cells increased nearly 60% when co-cultured with LNCaP (PSMA+), Wash Tee as compared with DU145 (PSMA-).Conclusion: Here, we describe the ability of nanobody-based CAR to recognize PSMA that leads to the activation of Jurkat cells.This construct might be used as a promising candidate for clinical applications in prostate cancer therapy.