1. Genetically engineered T cells bearing chimeric nanoconstructed receptors harboring TAG-72-specific camelid single domain antibodies as targeting agents
- Author
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Zahra Sharifzadeh, S. Moein Moghimi, Fatemeh Rahbarizadeh, Davoud Ahmadvand, Mohammad Ali Shokrgozar, Fereidoun Mahboudi, Fatemeh Rahimi Jamnani, Institut Pasteur d'Iran, Réseau International des Instituts Pasteur (RIIP), Department of Medical Biotechnology, Tarbiat Modares University [Tehran]-Faculty of Medical Sciences, School of Allied Medical Sciences, Tehran University of Medical Sciences, Biotechnology Research Center, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Nano-Science Center [Copenhagen], Faculty of Health and Medical Sciences, University of Copenhagen = Københavns Universitet (KU)-University of Copenhagen = Københavns Universitet (KU)-Faculty of Science [Copenhagen], University of Copenhagen = Københavns Universitet (KU), Nanomedicine Laboratory, Centre of Pharmaceutical Nanotechnology and Nanotoxicology-University of Copenhagen = Københavns Universitet (KU), and This paper was supported by Pasteur Institute of Iran, Tehran, Iran, the Biotechnology Committee of Tarbiat Modares University (TMU-88-8-67), Tehran, Iran and from the Danish Agency for Science, Technology and Innovative (det Strategiske Forskningsråd) reference 09-065746/DSF to SMM.
- Subjects
Cancer Research ,Leukemia, T-Cell ,medicine.medical_treatment ,T cell ,Recombinant Fusion Proteins ,T-Lymphocytes ,Biology ,Lymphocyte Activation ,Transfection ,Immunotherapy, Adoptive ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Antigen ,Antibody Specificity ,Antigens, Neoplasm ,Peptide Library ,medicine ,Animals ,Humans ,Chimeric antigen receptor ,030304 developmental biology ,Glycoproteins ,0303 health sciences ,Immunogenicity ,CD28 ,Immunotherapy ,Single-Domain Antibodies ,Molecular biology ,3. Good health ,Cell biology ,Oligoclonal single domain antibodies ,Receptors, Antigen ,medicine.anatomical_structure ,Oncology ,TAG-72 ,030220 oncology & carcinogenesis ,Immunoglobulin G ,Colonic Neoplasms ,biology.protein ,Carcinoma, Squamous Cell ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Antibody ,Genetic Engineering ,Camelids, New World ,HT29 Cells ,Signal Transduction - Abstract
International audience; Despite the preclinical success of adoptive therapy with T cells bearing chimeric nanoconstructed antigen receptors (CARs), certain limitations of this therapeutic approach such as the immunogenicity of the antigen binding domain, the emergence of tumor cell escape variants and the blocking capacity of soluble antigen still remain. Here, we address these issues using a novel CAR binding moiety based on the oligoclonal camelid single domain antibodies. A unique set of 13 single domain antibodies were selected from an immunized camel phage library based on their target specificity and binding affinity. A combination of these single domain antibodies was used to generate four tumor associated glycoprotein (TAG-72)-specific CARs harboring an identical antigen binding site, but with different signaling and spacer domains. Although all four CARs were functionally active against the TAG-72 expressing tumor cells, the combination of CD3ζ, OX40, CD28 as well as the CH3-CH2-hinge-hinge domains most efficiently triggered T cell activation. Importantly, CAR mediated functions were not blocked by the soluble TAG-72 antigen at a supraphysiological concentration. Our approach may have the potential to reverse multiple tumor immune evasion mechanisms, avoid CAR immunogenicity, and overcome problems in cancer gene therapy with engineered nanoconstructs.
- Published
- 2012