Blockade of LFA-1 augments in vitro differentiation of antigen-induced Foxp3⁺ Treg cells

J Immunol Methods. 2014 Dec 1:414:58-64. doi: 10.1016/j.jim.2014.07.012. Epub 2014 Aug 6.

Abstract

Adoptive transfer of antigen-specific, in vitro-induced Foxp3(+) Treg (iTreg) cells protects against autoimmune disease. To generate antigen-specific iTreg cells at high purity, however, remains a challenge. Whereas polyclonal T cell stimulation with anti-CD3 and anti-CD28 antibody yields Foxp3(+) iTreg cells at a purity of 90-95%, antigen-induced iTreg cells typically do not exceed a purity of 65-75%, even in a TCR-transgenic model. In a similar vein to thymic Treg cell selection, iTreg cell differentiation is influenced not only by antigen recognition and the availability of TGF-β but also by co-factors including costimulation and adhesion molecules. In this study, we demonstrate that blockade of the T cell integrin Leukocyte Function-associated Antigen-1 (LFA-1) during antigen-mediated iTreg cell differentiation augments Foxp3 induction, leading to approximately 90% purity of Foxp3(+) iTreg cells. This increased efficacy not only boosts the yield of Foxp3(+) iTreg cells, it also reduces contamination with activated effector T cells, thus improving the safety of adoptive transfer immunotherapy.

Keywords: Autoimmunity; Foxp3; Immunotherapy; LFA-1; Treg cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • Animals
  • Autoimmune Diseases / immunology*
  • Autoimmunity / immunology
  • CTLA-4 Antigen / immunology
  • Cell Differentiation / immunology
  • Cells, Cultured
  • Forkhead Transcription Factors / biosynthesis*
  • Interleukin-2 / immunology
  • Lymphocyte Function-Associated Antigen-1 / immunology*
  • Mice
  • Mice, Transgenic
  • Signal Transduction / immunology
  • T-Lymphocytes, Regulatory / cytology*
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / transplantation
  • Transforming Growth Factor beta / immunology

Substances

  • CTLA-4 Antigen
  • Forkhead Transcription Factors
  • Foxp3 protein, mouse
  • Interleukin-2
  • Lymphocyte Function-Associated Antigen-1
  • Transforming Growth Factor beta