Immune Netw.  2016 Dec;16(6):358-365. 10.4110/in.2016.16.6.358.

Induction of Indoleamine 2,3-dioxygenase by Pre-treatment with Poly(I:C) May Enhance the Efficacy of MSC Treatment in DSS-induced Colitis

Affiliations
  • 1Department of Internal Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea. ckmin@catholic.ac.kr
  • 2Department of Pathology, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.

Abstract

Mesenchymal stem cells (MSCs) have been used experimentally for treating inflammatory disorders, partly owing to their immunosuppressive properties. The goal of the study was to determine whether TLR ligands can enhance the therapeutic efficacy of bone marrow-derived MSCs for the treatment of inflammatory bowel disease. Mice (C57BL6) were administered with 4% dextran sulfate sodium (DSS) in drinking water for 7 days and injected with MSCs on days 1 and 3 following DSS ingestion. Our results demonstrated that among various TLR ligands, MSCs treated with polyinosinic-polycytidylic acid [poly(I:C)], which is a TLR3 ligand, more profoundly induced IDO, which is a therapeutically relevant immunosuppressive factor, without any observable phenotype change in vitro. The poly(I:C)-treated MSCs attenuated the pathologic severity of DSS-induced murine colitis when injected i.p. but not i.v. In summary, preconditioning MSCs with poly(I:C) might improve their efficacy in treating DSS-induced colitis, and this effect at least partly depends on the enhancement of their immunosuppressive activity through increasing their production of IDO.

Keyword

Mesenchymal stem cells; Dextran sulfate sodium (DSS)-induced colitis; Toll-like receptors; Poly(I:C); Indoleamine 2,3-dioxygenase; Intraperitoneal route

MeSH Terms

Animals
Colitis*
Dextran Sulfate
Drinking Water
Eating
In Vitro Techniques
Indoleamine-Pyrrole 2,3,-Dioxygenase*
Inflammatory Bowel Diseases
Ligands
Mesenchymal Stromal Cells
Mice
Phenotype
Poly I-C
Toll-Like Receptors
Dextran Sulfate
Drinking Water
Indoleamine-Pyrrole 2,3,-Dioxygenase
Ligands
Poly I-C
Toll-Like Receptors

Figure

  • Figure 1 Immunophenotypic analysis of murine mesenchymal stem cells (MSCs). MSCs were cultured with or without TLR ligands for 24 h. (A) The expression of immunophenotypic surface antigens on MSCs was detected by flow cytometric analysis. (B) Morphological changes were observed in MSCs. There were no significant differences among the treatment groups. Gray color represents the control isotypes. Red lines show antibody-specific staining.

  • Figure 2 Effects of IFN-γ and TLR ligands on IDO expression in murine MSCs. (A) mRNA from 24-h cultures of unstimulated, IFNγ-stimulated, and IFNγ+TLR ligand-stimulated MSCs were assayed by quantitative RT-PCR for IDO expression. **p<0.01 between IFNγ-stimulated MSCs and IFNγ+polyinosinic-polycytidylic acid [poly(I:C)]-stimulated MSCs. (B) Western blot analysis of the protein expression of IDO was performed on 48-h cultures. *p<0.05 compared with unstimulated MSCs. The data shown are mean±SEM values calculated from four separate experiments.

  • Figure 3 Effects of i.v. injection of MSCs in the dextran sulfate sodium (DSS)-induced colitis model. (A) Schematic outline of the DSS model. DSS was given to mice via drinking water for 7 days to induce colitis. Afterwards, unstimulated MSCs (DSS+MSCs) or stimulated MSCs (DSS+sti-MSCs) were administered into the DSS-treated mice through a tail vein injection. The same volume of saline was injected into other DSStreated mice as positive controls (DSS control). All the mice were sacrificed 9 days after DSS administration. (B) Body weight and (C) disease activity index scores were monitored daily. (D) Colon lengths were measured on day 9. n=4 for DSS control, n=6 for treatment group. *p<0.05, **p<0.01, and ***p<0.001 compared with DSS control.

  • Figure 4 Effect of i.p. injection of MSCs in DSS-induced colitis model. (A) Schematic outline of the DSS model. DSS was given to mice via drinking water for 7 days to induce colitis. Afterwards, unstimulated MSCs (DSS+MSC) or stimulated MSCs (DSS+sti-MSC) were i.p. injected. The same volume of saline was administered into other DSS-treated mice as positive controls (DSS control). All the mice were sacrificed 9 days after DSS administration. (B) Body weight and (C) disease activity index scores were monitored daily. (D) Colon lengths were measured on day 9. n=4 for DSS control, n=6 for treatment group. **p<0.01 compared with DSS control.

  • Figure 5 Colonic mRNA expression of IDO according to the injection route of MSCs. On day 9 of DSS administration, colon tissues were harvested from mice administered with MSCs by (A) i.v. or (B) i.p. injection to measure the expression of IDO by quantitative RT-PCR. The data shown are mean±SEM values calculated from two separate experiments. n=4 for DSS control, n=6 for treatment group. ns=non-significant (p>0.05), **p<0.01 compared with DSS control.


Reference

1. Tsagias N, Koliakos I, Karagiannis V, Eleftheriadou M, Koliakos GG. Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes. Transfus Med. 2011; 21:253–261.
Article
2. Liang L, Dong C, Chen X, Fang Z, Xu J, Liu M, Zhang X, Gu DS, Wang D, Du W, Zhu D, Han ZC. Human umbilical cord mesenchymal stem cells ameliorate mice trinitrobenzene sulfonic acid (TNBS)-induced colitis. Cell Transplant. 2011; 20:1395–1408.
Article
3. Chamberlain G, Fox J, Ashton B, Middleton J. Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells. 2007; 25:2739–2749.
Article
4. DelaRosa O, Lombardo E. Modulation of adult mesenchymal stem cells activity by toll-like receptors: implications on therapeutic potential. Mediators Inflamm. 2010; 2010:865601.
Article
5. English K, Barry FP, Field-Corbett CP, Mahon BP. IFN-gamma and TNF-alpha differentially regulate immunomodulation by murine mesenchymal stem cells. Immunol Lett. 2007; 110:91–100.
Article
6. English K, Ryan JM, Tobin L, Murphy MJ, Barry FP, Mahon BP. Cell contact, prostaglandin E(2) and transforming growth factor beta 1 play non-redundant roles in human mesenchymal stem cell induction of CD4+CD25(High) forkhead box P3+ regulatory T cells. Clin Exp Immunol. 2009; 156:149–160.
Article
7. Tipnis S, Viswanathan C, Majumdar AS. Immunosuppressive properties of human umbilical cord-derived mesenchymal stem cells: role of B7-H1 and IDO. Immunol Cell Biol. 2010; 88:795–806.
Article
8. Sato K, Ozaki K, Oh I, Meguro A, Hatanaka K, Nagai T, Muroi K, Ozawa K. Nitric oxide plays a critical role in suppression of T-cell proliferation by mesenchymal stem cells. Blood. 2007; 109:228–234.
Article
9. Nishiyama Y, Kataoka T, Yamato K, Taguchi T, Yamaoka K. Suppression of dextran sulfate sodium-induced colitis in mice by radon inhalation. Mediators Inflamm. 2012; 2012:239617.
Article
10. Lim JY, Kim BG, Moon SK, Min CK. Enhancement of graft-versus-leukemia effects by mesenchymal stem cells in mixed chimerisim after a murine non-myeloablative hematopoietic stem cell transplantation. Korean J Hematol. 2008; 43:219–231.
Article
11. Gaudio E, Taddei G, Vetuschi A, Sferra R, Frieri G, Ricciardi G, Caprilli R. Dextran sulfate sodium (DSS) colitis in rats: clinical, structural, and ultrastructural aspects. Dig Dis Sci. 1999; 44:1458–1475.
12. Pevsner-Fischer M, Morad V, Cohen-Sfady M, Rousso-Noori L, Zanin-Zhorov A, Cohen S, Cohen IR, Zipor D. Toll-like receptors and their ligands control mesenchymal stem cell functions. Blood. 2007; 109:1422–1432.
Article
13. Mastri M, Shah Z, McLaughlin T, Greene CJ, Baum L, Suzuki G, Lee T. Activation of Toll-like receptor 3 amplifies mesenchymal stem cell trophic factors and enhances therapeutic potency. Am J Physiol Cell Physiol. 2012; 303:C1021–C1033.
Article
14. Caplan AI, Correa D. The MSC: an injury drugstore. Cell Stem Cell. 2011; 9:11–15.
Article
15. Opitz CA, Litzenburger UM, Lutz C, Lanz TV, Tritschler I, Koppel A, Tolosa E, Hoberg M, Anderl J, Aicher WK, Weller M, Wick W, Platten M. Toll-like receptor engagement enhances the immunosuppressive properties of human bone marrow-derived mesenchymal stem cells by inducing indoleamine-2,3-dioxygenase-1 via interferon-beta and protein kinase. R Stem Cells. 2009; 27:909–919.
Article
16. Hare JM, Traverse JH, Henry TD, Dib N, Strumpf RK, Schulman SP, Gerstenblith G, DeMaria AN, Denktas AE, Gammon RS, Hermiller JB Jr, Reisman MA, Schaer GL, Sherman W. A randomized, double-blind, placebo-controlled, dose-escalation study of intravenous adult human mesenchymal stem cells (prochymal) after acute myocardial infarction. J Am Coll Cardiol. 2009; 54:2277–2286.
Article
17. Liotta F, Angeli R, Cosmi L, Fili L, Manuelli C, Frosali F, Mazzinghi B, Maggi L, Pasini A, Lisi V, Santarlasci V, Consoloni L, Angelotti ML, Romagnani P, Parronchi P, Krampera M, Maggi E, Romagnani S, Annunziato F. Toll-like receptors 3 and 4 are expressed by human bone marrow-derived mesenchymal stem cells and can inhibit their T-cell modulatory activity by impairing Notch signaling. Stem Cells. 2008; 26:279–289.
Article
Full Text Links
  • IN
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr