A foundation for universal T-cell based immunotherapy: T cells engineered to express a CD19-specific chimeric-antigen-receptor and eliminate expression of endogenous TCR

Blood. 2012 Jun 14;119(24):5697-705. doi: 10.1182/blood-2012-01-405365. Epub 2012 Apr 24.


Clinical-grade T cells are genetically modified ex vivo to express a chimeric antigen receptor (CAR) to redirect specificity to a tumor associated antigen (TAA) thereby conferring antitumor activity in vivo. T cells expressing a CD19-specific CAR recognize B-cell malignancies in multiple recipients independent of major histocompatibility complex (MHC) because the specificity domains are cloned from the variable chains of a CD19 monoclonal antibody. We now report a major step toward eliminating the need to generate patient-specific T cells by generating universal allogeneic TAA-specific T cells from one donor that might be administered to multiple recipients. This was achieved by genetically editing CD19-specific CAR(+) T cells to eliminate expression of the endogenous αβ T-cell receptor (TCR) to prevent a graft-versus-host response without compromising CAR-dependent effector functions. Genetically modified T cells were generated using the Sleeping Beauty system to stably introduce the CD19-specific CAR with subsequent permanent deletion of α or β TCR chains with designer zinc finger nucleases. We show that these engineered T cells display the expected property of having redirected specificity for CD19 without responding to TCR stimulation. CAR(+)TCR(neg) T cells of this type may potentially have efficacy as an off-the-shelf therapy for investigational treatment of B-lineage malignancies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adult
  • Antigen-Presenting Cells / immunology
  • Antigens, CD19 / immunology*
  • Antigens, Neoplasm / immunology
  • CD28 Antigens / metabolism
  • CD3 Complex / metabolism
  • Cells, Cultured
  • Endonucleases / metabolism
  • Epitopes / immunology*
  • Gene Knockout Techniques
  • Genetic Engineering*
  • Humans
  • Immunotherapy / methods*
  • K562 Cells
  • Lymphocyte Activation / immunology
  • Receptors, Antigen, T-Cell / immunology*
  • Recombinant Proteins / immunology*
  • T-Lymphocytes / immunology*
  • Zinc Fingers


  • Antigens, CD19
  • Antigens, Neoplasm
  • CD28 Antigens
  • CD3 Complex
  • Epitopes
  • Receptors, Antigen, T-Cell
  • Recombinant Proteins
  • Endonucleases