Chimeric Antigen Receptors Combining 4-1BB and CD28 Signaling Domains Augment PI3kinase/AKT/Bcl-XL Activation and CD8+ T Cell-Mediated Tumor Eradication

Mol Ther. 2010 Feb;18(2):413-20. doi: 10.1038/mt.2009.210. Epub 2009 Sep 22.

Abstract

To enhance the strength of activation afforded by tumor antigen-specific receptors, we investigated the effect of adding combined CD28 and 4-1BB costimulatory signaling domains to a chimeric antigen receptor (CAR) specific for prostate-specific membrane antigen (PSMA). Having transferred receptors encompassing the CD28, 4-1BB, and/or CD3zeta cytoplasmic domains in primary human CD8(+) T cells, we find that the P28BBz receptor, which includes all three signaling domains, is superior to receptors that only include one or two of these domains in promoting cytokine release, in vivo T-cell survival and tumor elimination following intravenous T-cell administration to tumor-bearing severe combined immunodeficient (SCID)/beige mice. Upon in vitro exposure to PSMA, the P28BBZ receptor-induced the strongest PI(3)Kinase/Akt activation and Bcl-X(L) expression, and the least apoptosis in transduced peripheral blood CD8(+) T cells. These findings further support the concept of integrating optimized costimulatory properties into recombinant antigen receptors to augment the survival and function of genetically targeted T cells within the tumor microenvironment.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Surface / immunology
  • CD28 Antigens / genetics
  • CD28 Antigens / metabolism*
  • CD8-Positive T-Lymphocytes / immunology*
  • Cells, Cultured
  • Flow Cytometry
  • Glutamate Carboxypeptidase II / immunology
  • Humans
  • Immunoblotting
  • Mice
  • Mice, SCID
  • NIH 3T3 Cells
  • Neoplasms / genetics
  • Neoplasms / therapy
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Proto-Oncogene Proteins c-akt / genetics
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism*
  • bcl-X Protein / genetics
  • bcl-X Protein / metabolism*

Substances

  • Antigens, Surface
  • CD28 Antigens
  • Tumor Necrosis Factor Receptor Superfamily, Member 9
  • bcl-X Protein
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • FOLH1 protein, human
  • Glutamate Carboxypeptidase II