Costimulation by chimeric antigen receptors revisited the T cell antitumor response benefits from combined CD28-OX40 signalling

Int J Cancer. 2011 Dec 15;129(12):2935-44. doi: 10.1002/ijc.25960. Epub 2011 Mar 29.


The therapeutic success of adoptive therapy with chimeric antigen receptor (CAR) engineered T cells depends on the appropriate costimulation of CD3ζ to induce full T cell activation. Costimulatory endodomains of the CD28 family are therefore fused with CD3ζ in a dual signalling CAR. Serious adverse events in two most recent trials; however, highlight the need to analyse in more detail the impact of each costimulatory endodomain on individual effector functions of redirected T cells. We therefore performed a thoroughly controlled side-by-side comparison of the most frequently used endodomains with respect to their impact on CD4(+) and CD8(+) T cell effector functions. CD28 reinforced T cell proliferation and is mandatory to induce IL-2. In the absence of added IL-2, CD28 and OX40 (CD137) but not 4-1BB (CD134) enhanced specific cytolysis. While CD28, 4-1BB and OX40 similarly improved pro-inflammatory cytokine secretion, OX40 most efficiently prevented activation induced cell death of CD62L(-) effector memory T cells. CD28 was superior to initiate the T cell response, OX40 and 4-1BB sustained the response in long term with OX40 being most effective. We consequently combined the beneficial functions in a 3rd generation CD28-OX40 CAR which substantially improved the antitumor response without loosing specificity.

MeSH terms

  • Animals
  • CD28 Antigens / metabolism*
  • Cell Line, Tumor
  • Cytotoxicity, Immunologic
  • DNA, Recombinant
  • Humans
  • Immunotherapy, Adoptive / methods*
  • Lymphocyte Activation / immunology
  • Mice
  • Neoplasm Transplantation
  • Neoplasms / immunology
  • Neoplasms / therapy*
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, OX40 / metabolism*
  • Signal Transduction / immunology
  • T-Lymphocytes / immunology


  • CD28 Antigens
  • DNA, Recombinant
  • Receptors, Antigen, T-Cell
  • Receptors, OX40