en Related Fields Related Fields مقاله کامل Full Length/Original Article <strong>Background</strong>: Recently, modification of T cells with chimeric 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. <strong>Methods</strong>: 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. <strong>Results</strong>: 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⁺), as compared with DU145 (PSMA^-). <strong>Conclusion</strong>: 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. Chimeric antigen receptor, Immunotherapy, Prostate cancer, PSMA 81 88 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-798&slc_lang=en&sid=1 Mahmoud Hassani No Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran Fatemeh Hajari Taheri No Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran Zahra Sharifzadeh 0000-0002-4419-0315 No Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran Arash Arashkia 0000-0002-3117-3217 No Department of Virology, Pasteur Institute of Iran, Tehran, Iran Jamshid Hadjati 0000-0002-8747-123X No Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Wytske M. van Weerden No Department of Urology, Erasmus Medical Center, Rotterdam, Netherlands Shahriyar Abdoli No Department of Virology, Pasteur Institute of Iran, Tehran, Iran Mohammad Hossein Modarressi 0000-0002-8203-5674 No Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran Mohsen Abolhassani 0000-0003-1340-5114 Yes Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran