Archives of Dermatological Research

, Volume 300, Issue 3, pp 115–124 | Cite as

Use of donor bone marrow mesenchymal stem cells for treatment of skin allograft rejection in a preclinical rat model

  • Paolo Sbano
  • Aldo Cuccia
  • Benedetta Mazzanti
  • Serena Urbani
  • Betti Giusti
  • Ilaria Lapini
  • Luciana Rossi
  • Rosanna Abbate
  • Giuseppina Marseglia
  • Genni Nannetti
  • Francesca Torricelli
  • Clelia Miracco
  • Alberto Bosi
  • Michele Fimiani
  • Riccardo Saccardi
Original Paper

Abstract

Recent studies indicate that mesenchymal stem cells (MSC) exhibit a degree of immune privilege due to their ability to suppress T cell mediated responses causing tissue rejection; however, the impact of allogeneic MSC in the setting of organ transplantation has been poorly investigated so far. The aim of our study was to evaluate the effect of intravenous donor MSC infusion for clinical tolerance induction in allogeneic skin graft transplantations in rats. MSC were isolated from Wistar rats and administered in Sprague-Dawley rats receiving Wistar skin graft with or without cyclosporine A (CsA). Graft biopsies were performed at day 10 post transplantation in all experimental groups for histological and gene expression studies. Intravenous infusion with donor MSC in CsA-treated transplanted rats resulted in prolongation of skin allograft survival compared to control animals. Unexpectedly, donor MSC infusion in immunocompetent rats resulted in a faster rejection as compared to control group. Cytokine expression analysis at the site of skin graft showed that CsA treatment significantly decreased pro-inflammatory cytokines IFN-γ and IL-2 and reduced TNF-α gene expression; however, the level of TNF-α is high in MSC-treated and not immunosuppressed rats. Results of our study in a rat tissue transplantation model demonstrated a possible immunogenic role for donor (allogeneic) MSC, confirming the need of adequate preclinical experimentation before clinical use.

Keywords

Mesenchymal stem cells Rat Skin transplantation CsA Cytokine expression 

Notes

Acknowledgments

The authors wish to thank Prof. F. Lolli for his assistance with the statistics.

References

  1. 1.
    Aggarwal S, Pittenger MF (2005) Human mesenchymal stem cells modulate allogeneic immune cell response. Blood 105:1815–1822PubMedCrossRefGoogle Scholar
  2. 2.
    Barber WH, Mankin JA, Laskow DA, Deierhoi MH, Julian BA, Curtis JJ, Diethelm AG (1991) Long-term results of a controlled prospective study with transfusion of donor-specific bone marrow in 57 cadaveric renal allograft recipients. Transplantation 1:70–75CrossRefGoogle Scholar
  3. 3.
    Bartholomew A, Sturgeon C, Siatskas M, Ferrer K, McIntosh K, Patil S, Hardy W, Devine S, Ucker D, Deans R, Moseley A, Hoffman R (2002) Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol 1:42–48CrossRefGoogle Scholar
  4. 4.
    Chung NG, Jeong DC, Park SJ, Choi BO, Cho B, Kim HK, Chun CS, Won JH, Han CW (2004) Cotransplantation of marrow stromal cells may prevent lethal graft-versus-host disease in major histocompatibility complex mismatched murine hematopoietic stem cell transplantation. Int J Hematol 4:370–376CrossRefGoogle Scholar
  5. 5.
    Demir Y, Ozmen S, Klimczak A, Mukherjee AL, Siemionow M (2004) Tolerance induction in composite facial allograft transplantation in the rat model. Plast Reconstr Surg 7:1790–1801CrossRefGoogle Scholar
  6. 6.
    Dey B, Sykes M, Spitzer TR (1998) Outcomes of recipients of both bone marrow and solid organ transplants. A review. Medicine (Baltimore) 5:355–369Google Scholar
  7. 7.
    Djouad F, Plence P, Bony C, Tropel P, Apparailly F, Sany J, Noel D, Jorgensen C (2003) Immunosuppressive effect of mesenchymal stem cells favors tumor growth in allogeneic animals. Blood 102:3837–3844PubMedCrossRefGoogle Scholar
  8. 8.
    Djouad F, Fritz V, Apparailly F, Louis-Plence P, Bony C, Sany J, Jorgensen C, Noel D (2005) Reversal of the immunosuppressive properties of mesenchymal stem cells by tumor necrosis factor alpha in collagen-induced arthritis. Arthritis Rheum 5:1595–1603CrossRefGoogle Scholar
  9. 9.
    Dobson KR, Reading L, Haberey M, Marine X, Scutt A (1999) Centrifugal isolation of bone marrow from bone: an improved method for the recovery and quantitation of bone marrow osteoprogenitor cells from rat tibiae and femurae. Calcif Tissue Int 5:411–413CrossRefGoogle Scholar
  10. 10.
    Eliopoulos N, Stagg J, Lejeune L, Pommey S, Galipeau J (2005) Allogeneic marrow stromal cells are immune rejected by MHC class I- and class II-mismatched recipient mice. Blood 13:4057–4065CrossRefGoogle Scholar
  11. 11.
    Fehr T, Sykes M (2004) Tolerance induction in clinical transplantation. Transpl Immunol 2:117–130CrossRefGoogle Scholar
  12. 12.
    Friedenstein AJ (1976) Precursor cells of mechanocytes. Int Rev Cytol 47:327–359PubMedCrossRefGoogle Scholar
  13. 13.
    Gaciong Z, Koziak K, Religa P, Lisiecka A, Morzycka-Michalik M, Rell K, Kozlowska-Boszko B, Lao M (1995) Increased expression of growth factors during chronic rejection of human kidney allograft. Transplant Proc 27:928–929PubMedGoogle Scholar
  14. 14.
    Grinnemo KH, Mansson A, Dellgren G, Klingberg D, Wardell E, Drvota V, Tammik C, Holgersson J, Ringden, Sylven C, Le Blanc K (2004) Xenoreactivity and engraftment of human mesenchymal stem cells transplanted into infarcted rat myocardium. J Thorac Cardiovasc Surg 5:1293–1300CrossRefGoogle Scholar
  15. 15.
    Heslan JM, Renaudin K, Thebault P, Josien R, Cuturi MC, Chiffoleau E (2006) New evidence for a role of allograft accommodation in long-term tolerance. Transplantation 82:1185–1193PubMedCrossRefGoogle Scholar
  16. 16.
    Inoue S, Popp FC, Koehl GE, Piso P, Schlitt HJ, Geisser EK, Dahlke MH (2006) Immunomodulatory effects of mesenchymal stem cells in a rat organ transplant model. Translantation 81:1589–1595CrossRefGoogle Scholar
  17. 17.
    Khanna AK, Cairns VR, Becker CG, Hosenpud JD (1998) TGF-beta: a link between immunosuppression, nephrotoxicity, and CsA. Transplant Proc 4:944–945CrossRefGoogle Scholar
  18. 18.
    Le Blanc K, Tammik L, Sundberg B, Haynesworth SE, Ringden O (2003) Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. Scand J Immunol 1:11–20CrossRefGoogle Scholar
  19. 19.
    Le Blanc K, Rasmusson I, Sundberg B, Gotherstrom C, Hassan M, Uzunel M, Ringden O (2004) Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. Lancet 9419:1439–1441CrossRefGoogle Scholar
  20. 20.
    Lim Yeon J, Jeun SS, Lee KJ, Oh JH, Kim SM, Park SI, Jeong CH, Kang SG (2006) Multiple stem cell traits of expanded rat bone marrow stromal cells. Exp Neurol 199:416–426CrossRefGoogle Scholar
  21. 21.
    Liu Y, Song J, Liu W, Wan Y, Chen X, Hu C (2003) Growth and differentiation of rat bone marrow stromal cells: does 5-azacytidine trigger their cardiomyogenic differentiation? Cardiovasc Res 58:460–468PubMedCrossRefGoogle Scholar
  22. 22.
    Maccario R, Moretta A, Cometa A, Montagna D, Comoli P, Locatelli F, Podesta M, Frassoni F (2005) Human mesenchymal stem cells and cyclosporin a exert a synergistic suppressive effect on in vitro activation of alloantigen-specific cytotoxic lymphocytes. Biol Blood Marrow Transplant 12:1031–1032CrossRefGoogle Scholar
  23. 23.
    Mannon RB (2006) Therapeutic targets in the treatment of allograft fibrosis. Am J Transplant 6:867–875PubMedCrossRefGoogle Scholar
  24. 24.
    Mathew JM, Garcia-Morales RO, Carreno M, Jin Y, Fuller L, Blomberg B, Cirocco R, Burke GW, Ciancio G, Ricordi C, Esquenazi V, Tzakis AG, Miller J (2003) Immune responses and their regulation by donor bone marrow cells in clinical organ transplantation. Transpl Immunol 3–4:307–321CrossRefGoogle Scholar
  25. 25.
    Meisel R, Zibert A, Laryea M, Gobel U, Daubener W, Dilloo D (2004) Human bone marrow stromal cells inhibit allogeneic T-cell responses by indoleamine 2,3-dioxygenase-mediated tryptophan degradation. Blood 12:4619–4621CrossRefGoogle Scholar
  26. 26.
    Monaco AP, Clark AW, Wood ML, Sahyoun AI, Codish SD, Brown RW (1976) Possible active enhancement of a human cadaver renal allograft with antilymphocyte serum (ALS) and donor bone marrow: case report of an initial attempt. Surgery 4:384–392Google Scholar
  27. 27.
    Nauta AJ, Westerhuis G, Kruisselbrink AB, Lurvink EG, Willemze R, Fibbe WE (2006) Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a non-myeloablative setting. Blood 6:2114–2120CrossRefGoogle Scholar
  28. 28.
    Petit F, Minns AB, Nazzal JA, Hettiaratchy SP, Lantieri LL, Randolph MA, Lee WP (2004) Prolongation of skin allograft survival after neonatal injection of donor bone marrow and epidermal cells. Plast Reconstr Surg 113:270–276PubMedCrossRefGoogle Scholar
  29. 29.
    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 5411:143–147CrossRefGoogle Scholar
  30. 30.
    Plain KM, Boyd R, Verma ND, Robinson CM, Tran GT, Hodgkinson SJ, Hall BM (2007) Transplant tolerance associated with a Th1 response and not broken by IL-4, IL-5, and TGF-beta blockade or Th1 cytokine administration. Transplantation 83:764–773PubMedCrossRefGoogle Scholar
  31. 31.
    Rasmusson I, Ringden O, Sundberg B, Le Blanc K (2005) Mesenchymal stem cells inhibit lymphocyte proliferation by mitogens and alloantigens by different mechanisms. Exp Cell Res 1:33–41CrossRefGoogle Scholar
  32. 32.
    Shapiro R, Starzl TE (1998) Bone marrow augmentation in renal transplant recipients. Transplant Proc 4:1371–1374CrossRefGoogle Scholar
  33. 33.
    Sharma VK, Bologa RM, Xu GP, Li B, Mouradian J, Wang J, Serur D, Rao V, Suthanthiran M (1996) Intragraft TGF-beta 1 mRNA: a correlate of interstitial fibrosis and chronic allograft nephropathy. Kidney Int 49:1297–1303PubMedCrossRefGoogle Scholar
  34. 34.
    Stagg J, Pommey S, Eliopoulos N, Galipeau J (2006) Interferon-gamma-stimulated marrow stromal cells: a new type of nonhematopoietic antigen-presenting cell. Blood 6:2570–2577CrossRefGoogle Scholar
  35. 35.
    Starzl TE, Demetris AJ, Trucco M, Murase N, Ricordi C, Ildstad S, Ramos H, Todo S, Tzakis A, Fung JJ (1993) Cell migration and chimerism after whole-organ transplantation: the basis of graft acceptance. Hepatology 6:1127–1152Google Scholar
  36. 36.
    Sudres M, Norol F, Trenado A, Gregoire S, Charlotte F, Levacher B, Lataillade JJ, Bourin P, Holy X, Vernant JP, Klatzmann D, Cohen JL (2006) Bone marrow mesenchymal stem cells suppress lymphocytes proliferation in vitro but fail to prevent graft-versus-host disease in mice. J Immunol 176:7761–7767PubMedGoogle Scholar
  37. 37.
    Taylor HR, Morris PJ (1970) Skin grafting in rats. Am J Med Technol 4:149–157Google Scholar
  38. 38.
    Tse WT, Pendleton JD, Beyer WM, Egalka MC, Guinan EC (2003) Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation 3:389–397CrossRefGoogle Scholar
  39. 39.
    Wood ML, Gozzo JJ, Monaco AP (1972) Use of antilymphocyte serum and bone marrow for production of immunological tolerance and enhancement: review and recent experiments. Transplant Proc 4:523–529PubMedGoogle Scholar
  40. 40.
    Zappia E, Casazza S, Pedemonte E, Benvenuto F, Bonanni I, Gerdoni E, Giunti D, Ceravolo A, Cazzanti F, Frassoni F, Mancardi G, Uccelli A (2005) Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy. Blood 5:1755–1761CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Paolo Sbano
    • 1
  • Aldo Cuccia
    • 1
  • Benedetta Mazzanti
    • 2
  • Serena Urbani
    • 2
  • Betti Giusti
    • 3
  • Ilaria Lapini
    • 3
  • Luciana Rossi
    • 3
  • Rosanna Abbate
    • 3
  • Giuseppina Marseglia
    • 3
  • Genni Nannetti
    • 3
  • Francesca Torricelli
    • 3
  • Clelia Miracco
    • 4
  • Alberto Bosi
    • 2
  • Michele Fimiani
    • 1
  • Riccardo Saccardi
    • 2
  1. 1.Department of Clinical Medicine and Immunological Sciences, Section of DermatologyUniversity of SienaSienaItaly
  2. 2.Hematology UnitCareggi HospitalFlorenceItaly
  3. 3.Department of Medical and Surgical Critical Care and Center of Research, Transfer and High Education, “DENOTHE”University of FlorenceFlorenceItaly
  4. 4.Section of Pathological Anatomy and Histology, Department of Human Pathology and Oncology, Policlinico Le ScotteUniversity of SienaSienaItaly

Personalised recommendations