In vitro generation of mature, naive antigen-specific CD8(+) T cells with a single T-cell receptor by agonist selection

Leukemia. 2014 Apr;28(4):830-41. doi: 10.1038/leu.2013.285. Epub 2013 Oct 4.

Abstract

Peripheral blood T cells transduced with a tumor-specific T-cell receptor (TCR) face problems of auto-reactivity and lack of efficacy caused by cross-pairing of exogenous and endogenous TCR chains, as well as short term in vivo survival due to activation and growth factor-induced differentiation. We here studied an alternative strategy for the efficient generation of naive CD8(+) T cells with a single TCR. TCR-transduced human postnatal thymus-derived and adult mobilized blood-derived hematopoietic progenitor cells (HPCs) were differentiated to CD4(+)CD8(+) double-positive T cells using OP9-Delta-like 1 (OP9-DL1) cultures. Addition of the agonist peptide induced double positive cells to cross-present the peptide, leading, in the absence of co-stimulation, to cell cycle arrest and differentiation into mature CD8(+) T cells. Comprehensive phenotypic, molecular and functional analysis revealed the generation of naive and resting CD8(+) T cells through a process similar to thymic positive selection. These mature T cells show a near complete inhibition of endogenous TCRA and TCRB rearrangements and express high levels of the introduced multimer-reactive TCR. Upon activation, specific cytokine production and efficient killing of tumor cells were induced. Using this strategy, large numbers of high-avidity tumor-specific naive T cells can be generated from readily available HPCs without TCR chain cross-pairing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Differentiation
  • Cell Line, Tumor
  • Child
  • Child, Preschool
  • Gene Rearrangement, T-Lymphocyte
  • Humans
  • Immunotherapy, Adoptive
  • Infant
  • Infant, Newborn
  • Receptors, Antigen, T-Cell / agonists
  • Receptors, Antigen, T-Cell / physiology*

Substances

  • Receptors, Antigen, T-Cell