Polyclonal Expansion of Human CD4+CD25+ Regulatory T Cells
Based on results from experimental animal models, the adoptive transfer of CD4+CD25+FOXP3+ regulatory T cells (Treg) is expected to be efficacious in treating autoimmune and inflammatory diseases, as well as in preventing alloresponses after solid organ or stem-cell transplantation. For potential clinical applications, large numbers of Treg cells in maximum purity will be required to avoid the risk of disease exacerbation by contaminating effector T cells. We have recently described methods for the efficient in vitro expansion of human Treg cells and identified CD4+CD25highCD45RA+ T cells as the ideal starting population for the generation of homogeneous and stable Treg cell products. Here, we provide detailed instructions for their identification, isolation, expansion, and functional characterization.
Key wordsImmunotherapy Tolerance Suppressor T cell Transplantation T-cell therapy
- 11.Trzonkowski P, Bieniaszewska M, Juscinska J, Dobyszuk A, Krzystyniak A, Marek N, Mysliwska J, and Hellmann A (2009) First-in-man clinical results of the treatment of patients with graft versus host disease with human ex vivo expanded CD4+CD25+CD127- T regulatory cells. Clin Immunol 133:22–6.PubMedCrossRefGoogle Scholar
- 18.Baron U, Floess S, Wieczorek G, Baumann K, Grutzkau A, Dong J, Thiel A, Boeld TJ, Hoffmann P, Edinger M, Turbachova I, Hamann A, Olek S, and Huehn J (2007) DNA demethylation in the human FOXP3 locus discriminates regulatory T cells from activated FOXP3(+) conventional T cells. Eur J Immunol 37:2378–89.PubMedCrossRefGoogle Scholar
- 20.Liu W, Putnam AL, Xu-Yu Z, Szot GL, Lee MR, Zhu S, Gottlieb PA, Kapranov P, Gingeras TR, de St Groth BF, Clayberger C, Soper DM, Ziegler SF, and Bluestone JA (2006) CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J Exp Med 203:1701–11.PubMedCrossRefGoogle Scholar
- 21.Seddiki N, Santner-Nanan B, Martinson J, Zaunders J, Sasson S, Landay A, Solomon M, Selby W, Alexander SI, Nanan R, Kelleher A, and Fazekas de St Groth B (2006) Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J Exp Med 203:1693–700.PubMedCrossRefGoogle Scholar