Robust and cost effective expansion of human regulatory T cells highly functional in a xenograft model of graft-versus-host disease

Haematologica. 2013 Apr;98(4):533-7. doi: 10.3324/haematol.2012.076430. Epub 2012 Dec 14.


The low frequency of naturally occurring regulatory T cells (nTregs) in peripheral blood and the suboptimal protocols available for their ex vivo expansion limit the development of clinical trials based on the adoptive transfer of these cells. We have, therefore, generated a simplified, robust and cost-effective platform for the large-scale expansion of nTregs using a gas permeable static culture flask (G-Rex) in compliance with Good Manufacturing Practice. More than 10(9) putative Tregs co-expressing CD25 and CD4 molecules (92 ± 5%) and FoxP3 (69 ± 19%) were obtained within 21 days of culture. Expanded Tregs showed potent regulatory activity in vitro (80 ± 13% inhibition of CD8(+) cell division) and in vivo (suppression or delay of graft-versus-host disease in a xenograft mouse model) indicating that the cost-effective and simplified production of nTregs we propose will facilitate the implementation of clinical trials based on their adoptive transfer.

Publication types

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

MeSH terms

  • Adoptive Transfer
  • Animals
  • CD2 Antigens / immunology
  • CD2 Antigens / metabolism
  • Cell Culture Techniques / economics
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Cell Proliferation*
  • Cost-Benefit Analysis
  • Flow Cytometry
  • Forkhead Transcription Factors / immunology
  • Forkhead Transcription Factors / metabolism
  • Graft vs Host Disease / immunology*
  • Graft vs Host Disease / therapy
  • Humans
  • Immunophenotyping
  • Interleukin Receptor Common gamma Subunit / deficiency
  • Interleukin Receptor Common gamma Subunit / genetics
  • Kaplan-Meier Estimate
  • Mice
  • Mice, Inbred NOD
  • Mice, Knockout
  • Mice, SCID
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • T-Lymphocytes, Regulatory / immunology*
  • T-Lymphocytes, Regulatory / metabolism
  • T-Lymphocytes, Regulatory / transplantation
  • Transplantation, Heterologous


  • CD2 Antigens
  • FOXP3 protein, human
  • Forkhead Transcription Factors
  • Il2rg protein, mouse
  • Interleukin Receptor Common gamma Subunit