Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer |
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Authors: | Mahmoud Hassani Fatemeh Hajari Taheri Zahra Sharifzadeh Arash Arashkia Jamshid Hadjati Wytske M van Weerden Mohammad Hossein Modarressi Mohsen Abolhassani |
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Institution: | 1. Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran;2. Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran;3. Department of Virology, Pasteur Institute of Iran, Tehran, Iran;4. Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;5. Department of Urology, Erasmus Medical Center, Rotterdam, The Netherlands |
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Abstract: | Adoptive transfer of T cells expressing chimeric antigen receptors (CARs) is considered to be a novel anticancer therapy. To date, in most cases, single-chain variable fragments (scFvs) of murine origin have been used in CARs. However, this structure has limitations relating to the potential immunogenicity of mouse antigens in humans and the relatively large size of scFvs. For the first time, we used camelid nanobody (VHH) to construct CAR T cells against prostate specific membrane antigen (PSMA). The nanobody against PSMA (NBP) was used to show the feasibility of CAR T cells against prostate cancer cells. T cells were transfected, and then the surface expression of the CAR T cells was confirmed. Then, the functions of VHH-CAR T cell were evaluated upon coculture with prostate cancer cells. At the end, the cytotoxicity potential of NBPII-CAR in T cells was approximated by determining the cell surface expression of CD107a after encountering PSMA. Our data show the specificity of VHH-CAR T cells against PSMA+ cells (LNCaP), not only by increasing the interleukin 2 (IL-2) cytokine (about 400 pg/mL), but also the expression of CD69 by almost 38%. In addition, VHH-CAR T cells were proliferated by nearly 60% when cocultured with LNCaP, as compared with PSMA negative prostate cancer cell (DU-145), which led to the upregulation of CD107a in T cells upto 31%. These results clearly show the possibility of using VHH-based CAR T cells for targeted immunotherapy, which may be developed to target virtually any tumor-associated antigen for adoptive T-cell immunotherapy of solid tumors. |
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Keywords: | chimeric antigen receptor T-cell immunotherapy prostate cancer prostate specific membrane antigen single-domain antibody fragment |
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