Frontline Science: Functionally impaired geriatric CAR-T cells rescued by increased α5β1 integrin expression

J Leukoc Biol. 2017 Aug;102(2):201-208. doi: 10.1189/jlb.5HI0716-322RR. Epub 2017 May 25.

Abstract

Chimeric antigen receptor expressing T cells (CAR-T) are a promising form of immunotherapy, but the influence of age-related immune changes on CAR-T production remains poorly understood. We showed that CAR-T cells from geriatric donors (gCAR-T) are functionally impaired relative to CAR-T from younger donors (yCAR-T). Higher transduction efficiencies and improved cell expansion were observed in yCAR-T cells compared with gCAR-T. yCAR-T demonstrated significantly increased levels of proliferation and signaling activation of phosphorylated (p)Erk, pAkt, pStat3, and pStat5. Furthermore, yCAR-T contained higher proportions of CD4 and CD8 effector memory (EM) cells, which are known to have enhanced cytolytic capabilities. Accordingly, yCAR-T demonstrated higher levels of tumor antigen-specific cytotoxicity compared with gCAR-T. Enhanced tumor killing by yCAR-T correlated with increased levels of perforin and granzyme B. yCAR-T had increased α5β1 integrin expression, a known mediator of retroviral transduction. We found that treatment with M-CSF or TGF-β1 rescued the impaired transduction efficiency of the gCAR-T by increasing the α5β1 integrin expression. Neutralization of α5β1 confirmed that this integrin was indispensable for CAR expression. Our study suggests that the increase of α5β1 integrin expression levels enhances CAR expression and thereby improves tumor killing by gCAR-T.

Keywords: M-CSF; TGF-β1; retronectin; transduction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adult
  • Aged
  • Aging / immunology*
  • Blotting, Western
  • Chimera
  • Cytotoxicity, Immunologic / immunology
  • Female
  • Flow Cytometry
  • Humans
  • Immunotherapy / methods*
  • Integrin alpha5beta1 / biosynthesis*
  • Integrin alpha5beta1 / immunology
  • Lymphocyte Activation / immunology
  • Male
  • Receptors, Antigen, T-Cell / immunology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • Young Adult

Substances

  • Integrin alpha5beta1
  • Receptors, Antigen, T-Cell