The role of Toll-like receptor 1 in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.06.022

Abstract

Abstract:

BACKGROUND: Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Toll-like receptor 1 in which will provide possible experimental basis for the future potential of cell therapy strategies.
OBJECTIVE: To investigate the role of Toll-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells.
METHODS: Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4⁺ T cells. CD4⁺ T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Toll-like receptor 1 expression levels related genes; number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4⁺ T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Toll-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry.
RESULTS AND CONCLUSION: Toll-like receptor 1 mRNA was expressed in both CD4⁺ T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4⁺ T cells than in CD4⁺ T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P < 0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Toll-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4⁺T cells compared with CD4⁺ T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P < 0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4⁺ T cells was significantly higher than that in CD4⁺ T cells cultured alone, (4.53±1.27)% vs. (2.39±0.80)% (P < 0.01). Toll-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: bone marrow, mesenchymal stem cells, Toll-Like receptor 1, Th17 cells, immunomodulation

CLC Number:

R394.2

Guo Zhen-xing, Chen Zhen-ping. The role of Toll-like receptor 1 in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(6): 956-961.

Figures/Tables 3

References

[1] Aggarwal S, Pittenger MF. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood. 2005; 105(4):1815-1822.

[2] 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(12):4619-4621.

[3] Glennie S, Soeiro I, Dyson PJ,et al. Bone marrow mesenchymal stem cells induce division arrest anergy of activated T cells.Blood. 2005;105(7):2821-2827.

[4] 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(2):386-398.

[5] English K, Mahon BP.Allogeneic mesenchymal stem cells: agents of immune modulation.J Cell Biochem. 2011;112(8): 1963-1968.

[6] 余劲明,蔡德鸿,张桦,等.同种异基因大鼠间充质干细胞移植对体内CD4⁺CD25⁺T细胞的影响[J].中国组织工程研究与临床康复, 2009,13(1):137-140.

[7] Jiang Y, Jahagirdar BN, Reinhardt RL,et al.Pluripotency of mesenchymal stem cells derived from adult marrow.Nature. 2002;418(6893):41-49.

[8] Patel SA, Sherman L, Munoz J, et al. Immunological properties of mesenchymal stem cells and clinical implications.Arch Immunol Ther Exp (Warsz). 2008;56(1):1-8.

[9] Le Blanc K, Rasmusson I, Sundberg B,et al.Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells.Lancet. 2004;363 (9419):1439-1441.

[10] Bartholomew A, Sturgeon C, Siatskas M,et al. Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo.Exp Hematol. 2002;30(1): 42-48.

[11] Zhang J, Li Y, Chen J,et al. Human bone marrow stromal cell treatment improves neurological functional recovery in EAE mice.Exp Neurol. 2005;195(1):16-26.

[12] Lazarus HM, Koc ON, Devine SM,et al.Cotransplantation of HLA-identical sibling culture-expanded mesenchymal stem cells and hematopoietic stem cells in hematologic malignancy patients.Biol Blood Marrow Transplant. 2005; 11(5):389-398.

[13] Guo Z, Zheng C, Chen Z, et al. FBM-derived mesenchymal stem cells promote the expansion of human Th17 cells, but inhibit the production of Th1 cells.Eur J Immunol. 2009; 39(10): 2840-2849.

[14] Schirmer C, Klein C, von Bergen M,et al. Human fibroblasts support the expansion of IL-17-producing T cells via up-regulation of IL-23 production by dendritic cells.Blood. 2010;116(10):1715-1725.

[15] Eljaafari A, Tartelin ML, Aissaoui H,et al. Bone marrow-derived and synovium-derived mesenchymal cells promote Th17 cell expansion and activation through caspase 1 activation: contribution to the chronicity of rheumatoid arthritis.Arthritis Rheum. 2012;64(7):2147-2157.

[16] Janeway CA Jr.Approaching the asymptote?Evolution and revolution in immunology.Cold Spring Harb Symp Quant Biol.1989;54 Pt 1:1-13.

[17] 陈青,张国梁,王勇强,等.TLR4对肝脏缺血再灌注损伤大鼠肝组织Th17细胞相关细胞因子表达的影响[J].山东医药,2013, 53(39):15-18.

[18] 李亭.Toll样受体在诱导小鼠小肠移植免疫耐受中的作用的实验研究[J].长沙:中南大学,2011.

[19] 钱雷,吕丽君,徐敏,等.活化的类风湿关节炎患者外周血单个核细胞Toll样受体4诱导Th17细胞分化[J].中华风湿病学杂志, 2011, 15(12):816-820.

[20] 李开艳.Th17细胞亚群在支气管哮喘中的发病机制及TLR2对Th17细胞分化的调控研究[J].武汉:华中科技大学,2011.

[21] Fang C,Zhang X,Miwa T.Complement promotes the development of inflammatory T-helper 17 cells through synergistic interaction with Toll-like receptor signaling and interleukin-6 production.Blood.2009;114(5):1005-1015.

[22] Fischer M, Ehlers M.Toll-like receptors in autoimmunity. Ann N Y Acad Sci. 2008;1143:21-34.

[23] Reynolds JM,Pappu BP,Peng J,et al.Toll-like receptor 2 signaling in CD4(+) T lymphocytes promotes T helper 17 responses and regulates the pathogenesis of autoimmune disease. Immunity. 2010;32(5):692-702.

[24] Nyirenda MH, Sanvito L, Darlington PJ,et al.TLR2 stimulation drives human naive and effector regulatory T cells into a Th17-like phenotype with reduced suppressive function.J Immunol. 2011;187(5):2278-2290.

[25] Gupta S, Aggarwal S, Rashanravan B,et al.Th1- and Th2-like cytokines in CD4+ and CD8+ T cells in autism.J Neuroimmunol. 1998;85(1):106-109.

[26] Medzhitov R, Preston-Hurlburt P, Janeway CA Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature. 1997;388(6640):394-397.

[27] Krishnan J, Selvarajoo K, Tsuchiya M,et al.Toll-like receptor signal transduction.Exp Mol Med. 2007;39(4):421-438.

[28] Liew FY, Xu D, Brint EK, et al.Negative regulation of toll-like receptor-mediated immune responses.Nat Rev Immunol. 2005;5(6):446-458.

[29] Kaisho T,Akira S.Toll-like receptor function and signaling.J Allergy Clin Immunol.2006;117(5):979-987.

[30] Summers SA, Steinmetz OM, Gan PY,et al.Toll-like receptor 2 induces Th17 myeloperoxidase autoimmunity while Toll-like receptor 9 drives Th1 autoimmunity in murine vasculitis. Arthritis Rheum. 2011;63(4):1124-1135.

[31] Allam JP, Duan Y, Winter J,et al.Tolerogenic T cells, Th1/Th17 cytokines and TLR2/TLR4 expressing dendritic cells predominate the microenvironment within distinct oral mucosal sites.Allergy. 2011;66(4):532-539.

[32] Teixeira-Coelho M, Cruz A, Carmona J,et al.TLR2 deficiency by compromising p19 (IL-23) expression limits Th 17 cell responses to Mycobacterium tuberculosis.Int Immunol. 2011; 23(2):89-96.

[33] Mu HH, Hasebe A, Van Schelt A, et al.Novel interactions of a microbial superantigen with TLR2 and TLR4 differentially regulate IL-17 and Th17-associated cytokines.Cell Microbiol. 2011;13(3):374-387.

[34] Dzopalic T, Dragicevic A, Vasilijic S,et al.Loxoribine, a selective Toll-like receptor 7 agonist, induces maturation of human monocyte-derived dendritic cells and stimulates their Th-1- and Th-17-polarizing capability.Int Immunopharmacol. 2010;10(11):1428-1433.

[35] Tournadre A, Lenief V, Miossec P.Expression of Toll-like receptor 3 and Toll-like receptor 7 in muscle is characteristic of inflammatory myopathy and is differentially regulated by Th1 and Th17 cytokines.Arthritis Rheum. 2010;62(7):2144- 2151.

[36] Kattah MG, Wong MT, Yocum MD,et al.Cytokines secreted in response to Toll-like receptor ligand stimulation modulate differentiation of human Th17 cells.Arthritis Rheum. 2008; 58(6):1619-1629.

[37] Aliahmadi E, Gramlich R, Grützkau A,et al.TLR2-activated human langerhans cells promote Th17 polarization via IL-1beta, TGF-beta and IL-23.Eur J Immunol. 2009; 39(5): 1221-1230.

[38] Kattah MG, Wong MT, Yocum MD,et al. Cytokines secreted in response to Toll-like receptor ligand stimulation modulate differentiation of human Th17 cells.Arthritis Rheum. 2008; 58(6):1619-1629.

[39] Lombardi V, Van Overtvelt L, Horiot S,et al. Human dendritic cells stimulated via TLR7 and/or TLR8 induce the sequential production of Il-10, IFN-gamma, and IL-17A by naive CD4⁺ T cells.J Immunol. 2009;182(6):3372-3379.

[40] Tanaka J, Watanabe N, Kido M,et al. Human TSLP and TLR3 ligands promote differentiation of Th17 cells with a central memory phenotype under Th2-polarizing conditions.Clin Exp Allergy. 2009;39(1):89-100.

[41] Marta M, Andersson A, Isaksson M,et al. Unexpected regulatory roles of TLR4 and TLR9 in experimental autoimmune encephalomyelitis.Eur J Immunol. 2008; 38(2): 565-575.

[42] Abdollahi-Roodsaz S, Joosten LA, Koenders MI,et al. Stimulation of TLR2 and TLR4 differentially skews the balance of T cells in a mouse model of arthritis.J Clin Invest. 2008;118(1):205-216.