CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors
A human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolat...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2013-01-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116301512 |
| _version_ | 1811276281532121088 |
|---|---|
| author | Vanesa Alonso-Camino David Sánchez-Martín Marta Compte Natalia Nuñez-Prado Rosa M Diaz Richard Vile Luis Alvarez-Vallina |
| author_facet | Vanesa Alonso-Camino David Sánchez-Martín Marta Compte Natalia Nuñez-Prado Rosa M Diaz Richard Vile Luis Alvarez-Vallina |
| author_sort | Vanesa Alonso-Camino |
| collection | DOAJ |
| description | A human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolate antibodies termed CARbodies that recognize antigens expressed on the tumor cell surface in a proof-of-principle system. After three rounds of activation-selection there was a clear repertoire restriction, with the emergence dominant clones. The CARbodies were purified from bacterial cultures as soluble and active proteins. Furthermore, to validate its potential application for adoptive cell therapy, human T cells were transduced with a LV encoding a second-generation costimulatory CAR (CARv2) bearing the selected CARbodies. Transduced human primary T cells expressed significant levels of the CARbodies-based CARv2 fusion protein on the cell surface, and importantly could be specifically activated, after stimulation with tumor cells. This approach is a promising tool for the generation of antibodies fully adapted to the display format (CAR) and the selection context (cell synapse), which could extend the scope of current adoptive cell therapy strategies with CAR-redirected T cells. |
| first_indexed | 2024-04-12T23:53:42Z |
| format | Article |
| id | doaj.art-7cba63e40ee34a57a9ba3f4d7bb52423 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-12T23:53:42Z |
| publishDate | 2013-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-7cba63e40ee34a57a9ba3f4d7bb524232022-12-22T03:11:36ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.19CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen ReceptorsVanesa Alonso-Camino0David Sánchez-Martín1Marta Compte2Natalia Nuñez-Prado3Rosa M Diaz4Richard Vile5Luis Alvarez-Vallina6Molecular Immunology Unit, Hospital Universitario Puerta de Hierro, Madrid, SpainMolecular Immunology Unit, Hospital Universitario Puerta de Hierro, Madrid, SpainMolecular Immunology Unit, Hospital Universitario Puerta de Hierro, Madrid, SpainMolecular Immunology Unit, Hospital Universitario Puerta de Hierro, Madrid, SpainDepartment of Molecular Medicine, Mayo Clinic, Rochester, Minnesota, USADepartment of Molecular Medicine, Mayo Clinic, Rochester, Minnesota, USAMolecular Immunology Unit, Hospital Universitario Puerta de Hierro, Madrid, SpainA human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolate antibodies termed CARbodies that recognize antigens expressed on the tumor cell surface in a proof-of-principle system. After three rounds of activation-selection there was a clear repertoire restriction, with the emergence dominant clones. The CARbodies were purified from bacterial cultures as soluble and active proteins. Furthermore, to validate its potential application for adoptive cell therapy, human T cells were transduced with a LV encoding a second-generation costimulatory CAR (CARv2) bearing the selected CARbodies. Transduced human primary T cells expressed significant levels of the CARbodies-based CARv2 fusion protein on the cell surface, and importantly could be specifically activated, after stimulation with tumor cells. This approach is a promising tool for the generation of antibodies fully adapted to the display format (CAR) and the selection context (cell synapse), which could extend the scope of current adoptive cell therapy strategies with CAR-redirected T cells.http://www.sciencedirect.com/science/article/pii/S2162253116301512adoptive cell therapyantibodychimeric antigen receptorrepertoire selectiontumor-associated antigens |
| spellingShingle | Vanesa Alonso-Camino David Sánchez-Martín Marta Compte Natalia Nuñez-Prado Rosa M Diaz Richard Vile Luis Alvarez-Vallina CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors Molecular Therapy: Nucleic Acids adoptive cell therapy antibody chimeric antigen receptor repertoire selection tumor-associated antigens |
| title | CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors |
| title_full | CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors |
| title_fullStr | CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors |
| title_full_unstemmed | CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors |
| title_short | CARbodies: Human Antibodies Against Cell Surface Tumor Antigens Selected From Repertoires Displayed on T Cell Chimeric Antigen Receptors |
| title_sort | carbodies human antibodies against cell surface tumor antigens selected from repertoires displayed on t cell chimeric antigen receptors |
| topic | adoptive cell therapy antibody chimeric antigen receptor repertoire selection tumor-associated antigens |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116301512 |
| work_keys_str_mv | AT vanesaalonsocamino carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT davidsanchezmartin carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT martacompte carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT natalianunezprado carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT rosamdiaz carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT richardvile carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors AT luisalvarezvallina carbodieshumanantibodiesagainstcellsurfacetumorantigensselectedfromrepertoiresdisplayedontcellchimericantigenreceptors |