1. An entirely humanized CD3 zeta-chain signaling receptor that directs peripheral blood t cells to specific lysis of carcinoembryonic antigen-positive tumor cells.
- Author
-
Hombach A, Schneider C, Sent D, Koch D, Willemsen RA, Diehl V, Kruis W, Bolhuis RL, Pohl C, and Abken H
- Subjects
- Antibody Specificity, Carcinoembryonic Antigen biosynthesis, Carrier Proteins biosynthesis, Carrier Proteins genetics, Colonic Neoplasms therapy, Cross Reactions, Cytotoxicity, Immunologic, Epitopes, T-Lymphocyte immunology, Gene Transfer Techniques, Humans, Immunoglobulin Fragments genetics, Immunoglobulin Fragments immunology, Immunomagnetic Separation, Immunotherapy, Adoptive methods, Leukemia Virus, Gibbon Ape genetics, Leukemia Virus, Murine genetics, Lymphocyte Activation, Membrane Proteins biosynthesis, Membrane Proteins genetics, Protein Structure, Tertiary, Receptor-CD3 Complex, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell biosynthesis, Receptors, Antigen, T-Cell genetics, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins immunology, Recombinant Proteins genetics, Recombinant Proteins immunology, Signal Transduction immunology, T-Lymphocytes metabolism, T-Lymphocytes virology, Carcinoembryonic Antigen immunology, Carrier Proteins immunology, Colonic Neoplasms immunology, Membrane Proteins immunology, Receptor-CD3 Complex, Antigen, T-Cell immunology, Receptors, Antigen, T-Cell immunology, Receptors, Cell Surface, T-Lymphocytes immunology
- Abstract
Recombinant T-cell receptors with antibody-like specificity for tumor-associated antigens are successfully used to direct the cytolytic activity of T cells toward tumor cells. Clinical application, however, needs to comply with the low immunogenicity of the recombinant receptor, efficient gene transfer into peripheral blood T cells, and enrichment of receptor-grafted cells. Here, we address these issues and describe an entirely humanized immune receptor for use in adoptive immunotherapy of colorectal carcinoma. The receptor consists of a single-chain antibody (scFv) binding domain specific for carcinoembryonic antigen (CEA), the IgG hinge and CH2/CH3 (Fc) joining region, and the transmembrane and intracellular CD3 zeta signaling chain. To express the receptor in peripheral blood T cells, both GALV envelope and MuLV 4070A pseudotyped retrovirus turned out to be equally efficient, with transduction efficiencies of about 5% to 40%, depending on the lymphocyte donor. Furthermore, receptor-grafted T cells could be 2- to 6-fold enriched by magnetic activated cell sorting, utilizing an antibody directed to the extracellular IgG domain of the receptor. Upon co-culture with CEA(+) tumor cells, receptor-grafted T cells are specifically and efficiently activated to cytolysis and IFN-gamma secretion, demonstrating their feasibility for the adoptive immunotherapy of CEA(+) carcinomas., (Copyright 2000 Wiley-Liss, Inc.)
- Published
- 2000