title: An entirely humanized CD3 zeta-chain signaling receptor that directs peripheral blood t cells to specific lysis of carcinoembryonic antigen-positive tumor cells.
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.
authors.affiliation: Klinik I für Innere Medizin, Labor Tumorgenetik, Universität zu Köln, Köln, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Hombach
authors.forename: C
authors.initials: C
authors.lastname: Schneider
authors.forename: D
authors.initials: D
authors.lastname: Sent
authors.forename: D
authors.initials: D
authors.lastname: Koch
authors.forename: R A
authors.initials: RA
authors.lastname: Willemsen
authors.forename: V
authors.initials: V
authors.lastname: Diehl
authors.forename: W
authors.initials: W
authors.lastname: Kruis
authors.forename: R L
authors.initials: RL
authors.lastname: Bolhuis
authors.forename: C
authors.initials: C
authors.lastname: Pohl
authors.forename: H
authors.initials: H
authors.lastname: Abken
mesh_terms: D000918:Antibody Specificity; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis / Q000276:immunology*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D003110:Colonic Neoplasms / Q000276:immunology* / Q000628:therapy; D003429:Cross Reactions; D003602:Cytotoxicity, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D018014:Gene Transfer Techniques; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D018189:Immunomagnetic Separation; D016219:Immunotherapy, Adoptive / Q000379:methods; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics; D009052:Leukemia Virus, Murine / Q000235:genetics; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D017434:Protein Structure, Tertiary; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000821:virology
chemical_list: D002272:Carcinoembryonic Antigen; D002352:Carrier Proteins; D018984:Epitopes, T-Lymphocyte; D007128:Immunoglobulin Fragments; D008565:Membrane Proteins; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D011948:Receptors, Antigen, T-Cell; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C054979:antigen T cell receptor, zeta chain; C065686:carcinoembryonic antigen binding protein, human