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Functional analysis and gene expression profile of umbilical cord blood regulatory T cells

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Abstract

The aim of the study was to analyze and compare the functional properties and the gene expression profile of regulatory T cells (Tregs) isolated from cord blood (CB) units (n = 23) and from the peripheral blood (PB) of adult normal donors (n = 13). Tregs were purified from mononuclear cells and expanded for 6 days with anti-CD3, anti-CD28, and IL-2. CB and PB Tregs presented similar immunophenotypic features. However, Tregs isolated from CB presented a much higher expansion capacity; this was confirmed by the genomic characterization that showed in CB-derived Tregs significant enrichments of genes involved in cell proliferation, chromatin modification, and regulation of gene expression. All samples were positive for the FoxP3 gene and protein after expansion. CB and PB expanded Tregs exerted a comparable and potent suppressive function on the proliferative reaction of autologous T cells stimulated by allogeneic dendritic cells and presented a high in vitro IL-10 production capacity. Gene profile analysis also revealed for PB Tregs significant enrichments of genes involved in the adaptive immune response. These data offer further insights into the understanding of the biology of CB transplantation indicating a possible role played by CB Tregs in the suppression of the allogeneic T cell response.

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References

  1. Kurtzberg J, Laughlin M, Graham ML, Smith C, Olson JF, Halperin EC et al (1996) Placental blood as a source of hematopoietic stem cells for transplantation into unrelated recipients. N Engl J Med 335:157–166

    Article  PubMed  CAS  Google Scholar 

  2. Gluckman E, Rocha V, Boyer-Chammard A, Locatelli F, Arcese W, Pasquini R et al (1997) Outcome of cord-blood transplantation from related and unrelated donors. Eurocord Transplant Group and the European Blood and Marrow Transplantation Group. N Engl J Med 337:373–381

    Article  PubMed  CAS  Google Scholar 

  3. Rubinstein P, Carrier C, Scaradavou A, Kurtzberg J, Adamson J, Migliaccio AR et al (1998) Outcomes among 562 recipients of placental-blood transplants from unrelated donors. N Engl J Med 339:1565–1577

    Article  PubMed  CAS  Google Scholar 

  4. Laughlin MJ, Barker J, Bambach B, Koc ON, Rizzieri DA, Wagner JE et al (2001) Hematopoietic engraftment and survival in adult recipients of umbilical-cord blood from unrelated donors. N Engl J Med 344:1815–1822

    Article  PubMed  CAS  Google Scholar 

  5. Wagner JE, Barker JN, DeFor TE, Baker KS, Blazar BR, Eide C et al (2002) Transplantation of unrelated donor umbilical cord blood in 102 patients with malignant and non-malignant diseases: influence of CD34 cell dose and HLA disparity on treatment-related mortality and survival. Blood 100:1611–1618

    PubMed  CAS  Google Scholar 

  6. Arcese W, Rocha V, Labopin M, Sanz G, Iori AP, de Lima M et al (2006) Unrelated cord blood transplants in adults with hematologic malignancies. Haematologica 91:223–230

    PubMed  Google Scholar 

  7. Iori AP, Arcese W, Milano F, Calabrese E, Torelli GF, Barberi W et al (2007) Unrelated cord blood transplant in children with high-risk acute lymphoblastic leukemia: a long-term follow-up. Haematologica 92:1051–1058

    Article  PubMed  Google Scholar 

  8. Wagner JE, Rosenthal J, Sweetman R, Shu XO, Davies SM, Ramsay NK et al (1996) Successful transplantation of HLA-matched and HLA-mismatched umbilical cord blood from unrelated donors: analysis of engraftment and acute graft-versus-host disease. Blood 88:795–802

    PubMed  CAS  Google Scholar 

  9. Locatelli F, Rocha V, Chastang C, Arcese W, Michel G, Abecasis M et al (1999) Factors associated with outcome after cord blood transplantation in children with acute leukemia. Eurocord–Cord Blood Transplant Group. Blood 93:3662–3671

    PubMed  CAS  Google Scholar 

  10. Rocha V, Wagner JE, Sobocinski KA, Klein JP, Zhang MJ, Horowitz MM et al (2000) Graft-versus-host disease in children who have received a cord-blood or bone marrow transplant from an HLA-identical sibling. Eurocord and International Bone Marrow Transplant Registry Working Committee on Alternative Donor and Stem Cell Source. N Engl J Med 342:1846–1854

    Article  PubMed  CAS  Google Scholar 

  11. Wilson CB, Westall J, Johnston L, Lewis DB, Dower SK, Alpert AR (1986) Decreased production of interferon-γ by human neonatal cells. Intrinsic and regulatory deficiencies. J Clin Invest 77:860–867

    Article  PubMed  CAS  Google Scholar 

  12. Chang M, Suen Y, Lee SM, Baly D, Buzby JS, Knoppel E et al (1994) Transforming growth factor-β 1, macrophage inflammatory protein-1 α, and interleukin-8 gene expression is lower in stimulated human neonatal compared with adult mononuclear cells. Blood 84:118–124

    PubMed  CAS  Google Scholar 

  13. Lee SM, Suen Y, Chang L, Bruner V, Qian J, Indes J et al (1996) Decreased interleukin-12 from activated cord vs adult peripheral blood mononuclear cells and upregulation of interferon-γ, natural killer, and lymphokine-activated killer activity by IL-12 in cord blood MNC. Blood 88:945–954

    PubMed  CAS  Google Scholar 

  14. Zola H, Fusco M, Weedon H, Macardle PJ, RidingsJ RDM (1996) Reduced expression of the interleukin-2-receptor γ chain on cord blood lymphocytes: relationship to functional immaturity of the neonatal immune response. Immunology 87:86–91

    PubMed  CAS  Google Scholar 

  15. Qian JX, Lee SM, Suen Y, Knoppel E, van de Ven C, Cairo MS (1997) Decreased IL-15 from activated cord versus adult peripheral blood mononuclear cells (MNC) and the effect of IL-15 in up-regulating anti-tumor immune activity and cytokine production in cord blood. Blood 90:3106–3117

    PubMed  CAS  Google Scholar 

  16. Thornton CA, Upham JW, Wikstrom ME, Holt BJ, White GP, Sharp MJ et al (2004) Functional maturation of CD4+CD25+CTLA4+CD45RA+ T regulatory cells in human neonatal T cell responses to environmental antigens/allergens. J Immunol 173:3084–3092

    PubMed  CAS  Google Scholar 

  17. Godfrey WR, Spoden DJ, Ge YG, Baker SR, Liu B, Levine BL et al (2005) Cord blood CD4+CD25+-derived T regulatory cell lines express FoxP3 protein and manifest potent suppressor function. Blood 105:750–758

    Article  PubMed  CAS  Google Scholar 

  18. Schubert LA, Jeffery E, Zhang Y, Ramsdell F, Ziegler SF (2001) Scurfin (FOXP3) acts as a repressor of transcription and regulates T cell activation. J Biol Chem 276:37672–37679

    Article  PubMed  CAS  Google Scholar 

  19. Nagler-Anderson C, Bhan AK, Podolsky DK, Terhorst C (2004) Control freaks: immune regulatory cells. Nat Immunol 5:119–122

    Article  PubMed  CAS  Google Scholar 

  20. Fehervari Z, Sakaguchi S (2004) CD4+ Tregs and immune control. J Clin Invest 114:1209–1217

    PubMed  CAS  Google Scholar 

  21. Fehervari Z, Sakaguchi S (2004) Development and function of CD25+CD4+ regulatory T cells. Curr Opin Immunol 16:203–208

    Article  PubMed  CAS  Google Scholar 

  22. Annacker O, Pimenta-Araujo R, Burlen-Defranoux O, Barbosa TC, Cumano A, Bandeira A (2001) CD25+ CD4+ T cells regulate the expansion of peripheral CD4 T cells through the production of IL-10. J Immunol 166:3008–3018

    PubMed  CAS  Google Scholar 

  23. Kingsley CI, Karim M, Bushell AR, Wood KJ (2002) CD25+CD4+ regulatory T cells prevent graft rejection: CTLA-4- and IL-10-dependent immunoregulation of alloresponses. J Immunol 168:1080–1086

    PubMed  CAS  Google Scholar 

  24. Taylor PA, Panoskaltsis-Mortari A, Swedin JM, Lucas PJ, Gress RE, Levine BL et al (2004) L-Selectinhi but not the L-selectinlo CD4+25+ T-regulatory cells are potent inhibitors of GVHD and BM graft rejection. Blood 104:3804–3812

    Article  PubMed  CAS  Google Scholar 

  25. Edinger M, Hoffmann P, Ermann J, Drago K, Fathman CG, Strober S et al (2003) CD4 + CD25+ regulatory T cells preserve graft-versus-tumor activity while inhibiting graft-versus-host disease after bone marrow transplantation. Nat Med 9:1144–1150

    Article  PubMed  CAS  Google Scholar 

  26. Miura Y (2004) Association of FoxP3 regulatory gene expression with graft-versus-host disease. Blood 104:2187–2193

    Article  PubMed  CAS  Google Scholar 

  27. Taylor PA, Lees CJ, Blazar BR (2002) The infusion of ex vivo activated and expanded CD4+CD25+ immune regulatory cells inhibits graft-versus-host disease lethality. Blood 99:3493–3499

    Article  PubMed  CAS  Google Scholar 

  28. Hoffmann P, Ermann J, Edinger M, Fathman CG, Strober S (2002) Donor-type CD4(+) CD25(+) regulatory T cells suppress lethal acute graft-versus-host-disease after allogeneic bone marrow transplantation. J Exp Med 196:389–399

    Article  PubMed  CAS  Google Scholar 

  29. Cohen JL, Trenado A, Vasey D, Klatzmann D, Salomon BL (2002) CD4+CD25+ immunoregulatory T cells: new therapeutics for graft-versus-host disease. J Exp Med 196:401–406

    Article  PubMed  CAS  Google Scholar 

  30. Trenado A, Charlotte F, Fisson S, Yagello M, Klatzmann D, Salomon BL et al (2003) Recipient-type specific CD4+CD25+ regulatory T cells favor immune reconstitution and control graft-versus-host disease while maintaining graft-versus-leukemia. J Clin Invest 112:1688–1696

    PubMed  CAS  Google Scholar 

  31. Di Ianni M, Falzetti F, Carotti A, Terenzi A, Castellino F, Bonifacio E et al (2011) Tregs prevent GvHD and promote immune reconstitution in HLA-haploidentical transplantation. Blood 117:3921–3928

    Article  PubMed  Google Scholar 

  32. Tolar J, Hippen KL, Blazar BR (2009) Immune regulatory cells in umbilical cord blood: T regulatory cells and mesenchymal stromal cells. Br J Haematol 147:200–206

    Article  PubMed  CAS  Google Scholar 

  33. Wing K, Ekmark A, Karlsson H, Rudin A, Suri-Payer E (2002) Characterization of human CD25+ T cells in thymus, cord and adult blood. Immunology 106:190–199

    Article  PubMed  CAS  Google Scholar 

  34. Wing K, Larsson P, Sandstrom K, Lundin SB, Suri-Payer E, Rudin A (2005) CD4+CD25+FOXP3+ regulatory T cells from human thymus and cord blood suppress antigen-specific T cell responses. Immunology 115:516–525

    Article  PubMed  CAS  Google Scholar 

  35. Wing K, Lindgren S, Kollberg G, Lundgren A, Harris RA, Rudin A et al (2003) CD4 T cell activation by myelin oligodendrocyte glycoprotein is suppressed by adult but not cord blood CD25+ T cells. Eur J Immunol 33:579–587

    Article  PubMed  CAS  Google Scholar 

  36. Chang C-C, Satwani P, Oberfiel N, Vlad G, Simpson LL, Cairo MS (2005) Increased induction of allogeneic-specific cord blood CD4+CD25+ regulatory T (Treg) cells: a comparative study of naïve and antigenic-specific cord blood Treg cells. Exp Hematol 33:1508–1520

    Article  PubMed  CAS  Google Scholar 

  37. Fujimaki W, Takahashi N, Ohnuma K, Nagatsu M, Kurosawa H, Yoshida S et al (2008) Comparative study of regulatory T cell function of human CD25CD4 T cells from thymocytes, cord blood, and adult peripheral blood. Clin Dev Immunol 2008:305859

    Article  PubMed  Google Scholar 

  38. Brunstein CG, Miller JS, Cao Q, McKenna DH, Hippen KL, Curtsinger J et al (2011) Infusion of ex vivo expanded T regulatory cells in adults transplanted with umbilical cord blood: safety profile and detection kinetics. Blood 117:1061–1070

    Article  PubMed  CAS  Google Scholar 

  39. Maggio R, Peragine N, Calabrese E, De Propris MS, Intoppa S, Della Starza I et al (2007) Generation of functional dendritic cells (DC) in adult acute lymphoblastic leukemia: rationale for a DC-based vaccination program for patients in complete hematological remission. Leuk Lymphoma 48:302–310

    Article  PubMed  CAS  Google Scholar 

  40. Li C, Wong WH (2001) Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc Natl Acad Sci USA 98:31–36

    Article  PubMed  CAS  Google Scholar 

  41. Shevach EM (2002) CD4+ CD25+ suppressor T cells: more questions than answers. Nat Rev Immunol 2:389–400

    PubMed  CAS  Google Scholar 

  42. Andersson J, Stefanova I, Stephens GL, Shevach EM (2007) CD4 + CD25+ regulatory T cells are activated in vivo by recognition of self. Int Immunol 19:557–566

    Article  PubMed  CAS  Google Scholar 

  43. Davidson TS, DiPaolo RJ, Andersson J, Shevach EM (2007) Cutting edge: IL-2 is essential for TGF-beta-mediated induction of Foxp3+ T regulatory cells. J Immunol 178:4022–4026

    PubMed  CAS  Google Scholar 

  44. Stephens GL, Andersson J, Shevach EM (2007) Distinct subsets of FoxP3+ regulatory T cells participate in the control of immune responses. J Immunol 178:6901–6911

    PubMed  CAS  Google Scholar 

  45. Hippen KL, Harker-Murray P, Porter SB, Merkel SC, Londer A, Taylor DK et al (2008) Umbilical cord blood regulatory T-cell expansion and functional effects of tumor necrosis factor receptor family members OX40 and 4-1BB expressed on artificial antigen-presenting cells. Blood 112:2847–2857

    Article  PubMed  CAS  Google Scholar 

  46. MacMillan ML, Weisdorf DJ, Brunstein CG, Cao Q, DeFor TE, Verneris MR et al (2009) Acute graft-versus-host disease after unrelated donor umbilical cord blood transplantation: analysis of risk factors. Blood 113:2410–2415

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study recieved research grant support from the Istituto Superiore di Sanità (ISS), Rome, n. 7OAF3; Progetto “Oncologia,” Ministero della Salute, Fondazione Italiana di Ricerca in Medicina Sperimentale (FIRMS), Turin; Compagnia di San Paolo, Turin; and Associazione Italiana per la Ricerca sul Cancro (AIRC), Milan, Italy.

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Authors and Affiliations

  1. Division of Hematology, Department of Cellular Biotechnologies and Hematology, “Sapienza” University of Rome, Via Benevento 6, 00161, Rome, Italy

    Giovanni Fernando Torelli, Roberta Maggio, Nadia Peragine, Sabina Chiaretti, Maria Stefania De Propris, Barbarella Lucarelli, Maria Grazia Mascolo, Anna Paola Iori, Anna Guarini & Robin Foà

  2. Cord Blood Bank, Department of Transfusional Medicine, “Sapienza” University of Rome, Rome, Italy

    Maria Screnci & Gabriella Girelli

  3. Cord Blood Transplant Program, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    Filippo Milano

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  1. Giovanni Fernando Torelli
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  2. Roberta Maggio
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Correspondence to Giovanni Fernando Torelli.

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Supplementary Table 1

List of genes differentially expressed by Tregs expanded from CB units and from normal donor PB. (PDF 237 kb)

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Torelli, G.F., Maggio, R., Peragine, N. et al. Functional analysis and gene expression profile of umbilical cord blood regulatory T cells. Ann Hematol 91, 155–161 (2012). https://doi.org/10.1007/s00277-011-1288-y

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