An entirely humanized CD3 zeta-chain signaling receptor that directs peripheral blood t cells to specific lysis of carcinoembryonic antigen-positive tumor cells

Int J Cancer. 2000 Oct 1;88(1):115-20.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Specificity
  • Carcinoembryonic Antigen / biosynthesis
  • Carcinoembryonic Antigen / immunology*
  • Carrier Proteins / biosynthesis
  • Carrier Proteins / genetics
  • Carrier Proteins / immunology*
  • Colonic Neoplasms / immunology*
  • 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
  • Membrane Proteins / immunology*
  • Protein Structure, Tertiary
  • Receptor-CD3 Complex, Antigen, T-Cell / genetics
  • Receptor-CD3 Complex, Antigen, T-Cell / immunology*
  • Receptors, Antigen, T-Cell / biosynthesis
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / immunology*
  • Receptors, Cell Surface*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Signal Transduction / immunology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • T-Lymphocytes / virology


  • Carcinoembryonic Antigen
  • Carrier Proteins
  • Epitopes, T-Lymphocyte
  • Immunoglobulin Fragments
  • Membrane Proteins
  • Receptor-CD3 Complex, Antigen, T-Cell
  • Receptors, Antigen, T-Cell
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • antigen T cell receptor, zeta chain
  • carcinoembryonic antigen binding protein, human