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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibody Specificity
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Carcinoembryonic Antigen / biosynthesis
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Carcinoembryonic Antigen / immunology*
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Carrier Proteins / biosynthesis
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Carrier Proteins / genetics
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Carrier Proteins / immunology*
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Colonic Neoplasms / immunology*
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Colonic Neoplasms / therapy
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Cross Reactions
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Cytotoxicity, Immunologic
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Epitopes, T-Lymphocyte / immunology
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Gene Transfer Techniques
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Humans
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Immunoglobulin Fragments / genetics
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Immunoglobulin Fragments / immunology
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Immunomagnetic Separation
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Immunotherapy, Adoptive / methods
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Leukemia Virus, Gibbon Ape / genetics
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Leukemia Virus, Murine / genetics
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Lymphocyte Activation
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Membrane Proteins / biosynthesis
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Membrane Proteins / genetics
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Membrane Proteins / immunology*
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Protein Structure, Tertiary
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Receptor-CD3 Complex, Antigen, T-Cell / genetics
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Receptor-CD3 Complex, Antigen, T-Cell / immunology*
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Receptors, Antigen, T-Cell / biosynthesis
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Receptors, Antigen, T-Cell / genetics
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Receptors, Antigen, T-Cell / immunology*
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Receptors, Cell Surface*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Signal Transduction / immunology
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T-Lymphocytes / immunology*
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T-Lymphocytes / metabolism
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T-Lymphocytes / virology
Substances
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Carcinoembryonic Antigen
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Carrier Proteins
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Epitopes, T-Lymphocyte
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Immunoglobulin Fragments
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Membrane Proteins
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Receptor-CD3 Complex, Antigen, T-Cell
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Receptors, Antigen, T-Cell
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Receptors, Cell Surface
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Recombinant Fusion Proteins
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Recombinant Proteins
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antigen T cell receptor, zeta chain
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carcinoembryonic antigen binding protein, human