Immunosuppressive function of mesenchymal stem cells from human umbilical cord matrix in immune thrombocytopenia patients

Summary

Human umbilical cord matrix/Wharton's Jelly (hUC)-derived mesenchymal stem cells (MSC) have been shown to have marked therapeutic effects in a number of inflammatory diseases and autoimmune diseases in humans based on their potential for immunosuppression and their low immunogenicity. Currently, no data are available on the effectiveness of UC-MSC transplantation in immune thrombocytopenia (ITP) patients. It was the objective of this study to assess the effect of allogeneic UC-MSCs on ITP patients in vitro and in vivo. Peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BM-MNCs) from ITP patients and healthy controls were co-cultured with UC-MSCs for three days and seven days, respectively. Flow cytometry and ELISA were applied to assess the various parameters. In PBMCs from ITP patients, the proliferation of autoreactive T, B lymphocytes and destruction of autologous platelets were dramatically suppressed by UC-MSCs. UC-MSCs not only suppressed co-stimulatory molecules CD80, CD40L and FasL expression but also in shifting Th1/Th2/Treg cytokines profile in ITP patients. UC-MSCs obviously reversed the dysfunctions of megakaryocytes by promoting platelet production and decreasing the number of living megakaryocytes as well as early apoptosis. In addition, the level of thrombopoietin was increased significantly. Our clinical study showed that UC-MSCs play a role in alleviating refractory ITP by increasing platelet numbers. These findings suggested that UC-MSCs transplantation might be a potential therapy for ITP.

Keywords

UC-MSC - ITP - lymphocytes - platelets - megakaryocytes

• References

• 1 McMillan R. The pathogenesis of chronic immune (idiopathic) thrombocytopenic purpura. Semin Hematol 2000; 37 1 Suppl 1 5-9.
• 2 Semple JW. Infections, antigen-presenting cells, T cells, and immune tolerance: their role in the pathogenesis of immune thrombocytopenia. Hematol Oncol Clinics North Am 2009; 23: 1177-1192.
• 3 Yu J, Heck S, Patel V. et al. Defective circulating CD25 regulatory T cells in patients with chronic immune thrombocytopenic purpura. Blood 2008; 112: 1325-1328.
• 4 Semple JW, Milev Y, Cosgrave D. et al. Differences in serum cytokine levels in acute and chronic autoimmune thrombocytopenic purpura: relationship to platelet phenotype and antiplatelet T-cell reactivity. Blood 1996; 87: 4245-4254.
• 5 Wang T, Zhao H, Ren H. et al. Type 1 and type 2 T-cell profiles in idiopathic thrombocytopenic purpura. Haematologica 2005; 90: 914-923.
• 6 Panitsas F P, Theodoropoulou M, Kouraklis A. et al. Adult chronic idiopathic thrombocytopenic purpura (ITP) is the manifestation of a type-1 polarized immune response. Blood 2004; 103: 2645-2647.
• 7 Olsson B, Andersson PO, Jernas M. et al. T-cell-mediated cytotoxicity toward platelets in chronic idiopathic thrombocytopenic purpura. Nat Med 2003; 09: 1123-1124.
• 8 Zhang F, Chu X, Wang L. et al. Cell-mediated lysis of autologous platelets in chronic idiopathic thrombocytopenic purpura. Eur J Haematol 2006; 76: 427-431.
• 9 Deng J, Petersen BE, Steindler DA. et al. Mesenchymal stem cells spontaneously express neural proteins in culture and are neurogenic after transplantation. Stem Cells 2006; 24: 1054-1064.
• 10 Lee OK, Kuo TK, Chen WM. et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood 2004; 103: 1669-1675.
• 11 Pittenger MF, Mackay AM, Beck SC. et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143-147.
• 12 Sato Y, Araki H, Kato J. et al. Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. Blood 2005; 106: 756-763.
• 13 Stenderup K, Justesen J, Clausen C. et al. Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone 2003; 33: 919-926.
• 14 Rao MS, Mattson M P. Stem cells and aging: expanding the possibilities. Mech Ageing Develop 2001; 122: 713-734.
• 15 Deuse T, Stubbendorff M, Tang-Quan K. et al. Immunogenicity and immunomodulatory properties of umbilical cord lining mesenchymal stem cells. Cell Transplant 2011; 20: 655-667.
• 16 Romanov YA, Svintsitskaya VA, Smirnov VN. Searching for alternative sources of postnatal human mesenchymal stem cells: candidate MSC-like cells from umbilical cord. Stem Cells 2003; 21: 105-110.
• 17 Kim JW, Kim SY, Park SY. et al. Mesenchymal progenitor cells in the human umbilical cord. Ann Hematol 2004; 83: 733-738.
• 18 Fauza D. Amniotic fluid and placental stem cells. Best Pract Res 2004; 18: 877-891.
• 19 Sun L, Wang D, Liang J. et al. Umbilical cord mesenchymal stem cell transplantation in severe and refractory systemic lupus erythematosus. Arthritis Rheuma 2010; 62: 2467-2475.
• 20 Liu Y, Mu R, Wang S. et al. Therapeutic potential of human umbilical cord mesenchymal stem cells in the treatment of rheumatoid arthritis. Arthritis Res Therapy 2010; 12: R210.
• 21 Zappia E, Casazza S, Pedemonte E. et al. Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy. Blood 2005; 106: 1755-1761.
• 22 Provan D, Stasi R, Newland AC. et al. International consensus report on the investigation and management of primary immune thrombocytopenia. Blood 2009; 115: 168-186.
• 23 Rodeghiero F, Stasi R, Gernsheimer T. et al. Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children: report from an international working group. Blood 2009; 113: 2386-2393.
• 24 Lu LL, Liu YJ, Yang SG. et al. Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica 2006; 91: 1017-1026.
• 25 Jones S, Horwood N, Cope A. et al. The antiproliferative effect of mesenchymal stem cells is a fundamental property shared by all stromal cells. J Immunol 2007; 179: 2824-2831.
• 26 Cui G, Liu X, Yao J. The effect of costimulatory factors in the pathogenesis of chronic idiopathic thrombocytopenic purpura. J Huazhong Univ Science Technol Med Sci 2003; 23: 352-355.
• 27 Branehog I, Kutti J, Ridell B. et al. The relation of thrombokinetics to bone marrow megakaryocytes in idiopathic thrombocytopenic purpura (ITP). Blood 1975; 45: 551-562.
• 28 Faulhaber M, Wormann B, Ganser A. et al. In vitro response of myelodysplastic megakaryocytopoiesis to megakaryocyte growth and development factor (MGDF). Ann Hematol 2002; 81: 695-700.
• 29 Tajika K, Nakamura H, Nakayama K. et al. Thrombopoietin can influence mature megakaryocytes to undergo further nuclear and cytoplasmic maturation. Exp Hematol 2000; 28: 203-209.
• 30 Bernardo ME, Locatelli F, Fibbe WE. Mesenchymal stromal cells. Ann NY Acad Sci 2009; 1176: 101-117.
• 31 Krampera M, Cosmi L, Angeli R. et al. Role for interferon-gamma in the immunomodulatory activity of human bone marrow mesenchymal stem cells. Stem Cells 2006; 24: 386-398.
• 32 Ren G, Zhang L, Zhao X. et al. Mesenchymal stem cell-mediated immunosuppression occurs via concerted action of chemokines and nitric oxide. Cell Stem Cell 2008; 02: 141-150.
• 33 Semple J W, Provan D, Garvey MB. et al. Recent progress in understanding the pathogenesis of immune thrombocytopenia. Curr Opin Hematol 2010; 17: 590-595.
• 34 Wang M, Yang Y, Yang D. et al. The immunomodulatory activity of human umbilical cord blood-derived mesenchymal stem cells in vitro. Immunology 2009; 126: 220-232.
• 35 Di Nicola M, Carlo-Stella C, Magni M. et al. Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 2002; 99: 3838-3843.
• 36 Meisel R, Zibert A, Laryea M. et al. Human bone marrow stromal cells inhibit allogeneic T-cell responses by indoleamine 2,3-dioxygenase-mediated tryptophan degradation. Blood 2004; 103: 4619-4621.
• 37 Aggarwal S, Pittenger MF. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 2005; 105: 1815-1822.
• 38 Sato K, Ozaki K, Oh I. et al. Nitric oxide plays a critical role in suppression of T-cell proliferation by mesenchymal stem cells. Blood 2007; 109: 228-234.
• 39 Wang D, Chen K, Du WT. et al. CD14+ monocytes promote the immunosuppressive effect of human umbilical cord matrix stem cells. Exp Cell Res 2010; 316: 2414-2423.
• 40 Chen K, Wang D, Du WT. et al. Human umbilical cord mesenchymal stem cells hUC-MSCs exert immunosuppressive activities through a PGE2-dependent mechanism. Clin Immunol 2010; 135: 448-458.
• 41 Chang TT, Jabs C, Sobel RA. et al. Studies in B7-deficient mice reveal a critical role for B7 costimulation in both induction and effector phases of experimental autoimmune encephalomyelitis. J Exp Med 1999; 190: 733-740.
• 42 Peterson KE, Sharp GC, Tang H. et al. B7.2 has opposing roles during the activation versus effector stages of experimental autoimmune thyroiditis. J Immunol 1999; 162: 1859-1867.
• 43 Peng J, Liu C, Liu D. et al. Effects of B7-blocking agent and/or CsA on induction of platelet-specific T-cell anergy in chronic autoimmune thrombocytopenic purpura. Blood 2003; 101: 2721-2726.
• 44 Kuwana M, Okazaki Y, Kaburaki J. et al. Spleen is a primary site for activation of platelet-reactive T and B cells in patients with immune thrombocytopenic purpura. J Immunol 2002; 168: 3675-3682.
• 45 Van Kooten C, Banchereau J. CD40-CD40 ligand: a multifunctional receptor-ligand pair. Advances Immunol 1996; 61: 1-77.
• 46 Solanilla A, Pasquet JM, Viallard JF. et al. Platelet-associated CD154 in immune thrombocytopenic purpura. Blood 2005; 105: 215-218.
• 47 Batten P, Sarathchandra P, Antoniw JW. et al. Human mesenchymal stem cells induce T cell anergy and downregulate T cell allo-responses via the TH2 pathway: relevance to tissue engineering human heart valves. Tissue Engin 2006; 12: 2263-2273.
• 48 Selmani Z, Naji A, Zidi I. et al. Human leukocyte antigen-G5 secretion by human mesenchymal stem cells is required to suppress T lymphocyte and natural killer function and to induce CD4+CD25highFOXP3+ regulatory T cells. Stem Cells 2008; 26: 212-222.
• 49 Steller H. Mechanisms and genes of cellular suicide. Science 1995; 267: 1445-1449.
• 50 Zauli G, Vitale M, Falcieri E. et al. In vitro senescence and apoptotic cell death of human megakaryocytes. Blood 1997; 90: 2234-2243.
• 51 Falcieri E, Bassini A, Pierpaoli S. et al. Ultrastructural characterization of maturation, platelet release, and senescence of human cultured megakaryocytes. Anatom Rec 2000; 258: 90-99.
• 52 Houwerzijl EJ, Blom NR, van der Want JJ. et al. Megakaryocytic dysfunction in myelodysplastic syndromes and idiopathic thrombocytopenic purpura is in part due to different forms of cell death. Leukemia 2006; 20: 1937-1942.
• 53 Yang L, Wang L, Zhao CH. et al. Contributions of TRAIL-mediated megakaryocyte apoptosis to impaired megakaryocyte and platelet production in immune thrombocytopenia. Blood 2010; 116: 4307-4316.
• 54 De Botton S, Sabri S, Daugas E. et al. Platelet formation is the consequence of caspase activation within megakaryocytes. Blood 2002; 100: 1310-1317.
• 55 Crist SA, Elzey BD, Ludwig AT. et al. Expression of TNF-related apoptosis-inducing ligand (TRAIL) in megakaryocytes and platelets. Exp Hematol 2004; 32: 1073-1081.
• 56 Melloni E, Secchiero P, Celeghini C. et al. Functional expression of TRAIL and TRAIL-R2 during human megakaryocytic development. J Cell Physiol 2005; 204: 975-982.
• 57 Bacigalupo A, Valle M, Podesta M. et al. T-cell suppression mediated by mesenchymal stem cells is deficient in patients with severe aplastic anemia. Exp Hematol 2005; 33: 819-827.
• 58 Perez-Simon JA, Tabera S, Sarasquete ME. et al. Mesenchymal stem cells are functionally abnormal in patients with immune thrombocytopenic purpura. Cytotherapy 2009; 11: 1-8.
• 59 Liu B, Zhao H, Poon MC. et al. Abnormality of CD4(+)CD25(+) regulatory T cells in idiopathic thrombocytopenic purpura. Eur J Haematol 2007; 78: 139-143.