Up-regulation of CD200/CD200R1 Immunomodulatory Axis of Allogenic Peripheral Blood Mononuclear Cells in a Co-culture with Adipose-derived Mesenchymal Stem Cells
Co-inhibitory molecules modulate immune responses. Immunomodulatory properties of mesenchymal stem cells (MSCs) turn them into ideal candidates for cell therapy. This study was designed to investigate the immunomodulatory effect of adipose-derived stem cells (ASCs) on inflammatory environment of a co-culture of allogenic peripheral blood mononuclear cells (PBMCs) in a two-way mixed leukocyte reaction (twMLR) setting.
ASCs were co-cultured with allogenic PBMCs in twMLR setting for four days. The proliferation of peripheral blood mononuclear cells (PBMCs), levels of interleukin (IL)-10, and expression of interferon-gamma (IFN-γ), B7-1, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed death-ligand 1 (PD-L1), +, and CD200R1 genes, as well as cell surface expression of CD200 and CD200R1, were measured in twMLR as control, and co-culture groups on days 0, 2 and 4 of the experiment.
The proliferation of PBMCs was suppressed on days 2 and 4 of co-culture. The expression of CD200 (p=0.014), CD200R1, CTLA-4, and PD1 genes increased on days 2 and 4 of the co-culture compared to twMLR. CD200 expressing PBMCs decreased by 1.75% on day 2 of the co-culture but increased by 6.23% on day 4 of the co-culture (p=0.013) compared to the same days of twMLR. IL-10 levels increased in the co-culture supernatants on days 2 and 4 compared to twMLR (p<0.05).
Our results showed that ASCs upregulate the CD200/CD200R1 axis more than PD-1/PD-L1 and CTLA-4/B7-1 pathways in the twMLR. Also, elevated expression of CD200R1 in the final day of co-culture was similar to PD-1 expression pattern. This finding suggests a role for the CD200/CD200R1 axis in later modulation of the immune response.
2. Fontaine MJ, Shih H, Schäfer R, Pittenger MF. Unraveling the mesenchymal stromal cells' paracrine immunomodulatory effects. Transfus Med Rev. 2016;30(1):37-43.
3. Wang Y, Chen X, Cao W, Shi Y. Plasticity of mesenchymal stem cells in immunomodulation: pathological and therapeutic implications. Nat Immunol. 2014;15(11):1009.
4. Ren G, Zhang L, Zhao X, Xu G, Zhang Y, Roberts AI, et al. Mesenchymal stem cell-mediated immunosuppression occurs via concerted action of chemokines and nitric oxide. Cell stem cell. 2008;2(2):141-50.
5. Coulson-Thomas VJ, Coulson-Thomas YM, Gesteira TF, Kao WW-Y. Extrinsic and intrinsic mechanisms by which mesenchymal stem cells suppress the immune system. Ocul Sur. 2016;14(2):121-34.
6. Zhao Z-G, Xu W, Sun L, You Y, Li F, Li Q-B, et al. Immunomodulatory function of regulatory dendritic cells induced by mesenchymal stem cells. Immunol invest. 2012;41(2):183-98.
7. Coulson-Thomas VJ, Gesteira TF, Hascall V, Kao W. Umbilical cord mesenchymal stem cells suppress host rejection the role of the glycocalyx. J Biol Chem. 2014;289(34):23465-81.
8. Hoogduijn MJ. Are mesenchymal stromal cells immune cells? Arthritis Res Ther. 2015;17(1):88.
9. Mak TW, Saunders ME. The immune response : basic and clinical principles Amsterdam; Boston: Elsevier/Academic; 2006. Available from: http://site.ebrary.com/id/10188220.
10. Zhou J, He W, Luo G, Wu J. Mixed lymphocyte reaction induced by multiple alloantigens and the role for IL-10 in proliferation inhibition. Burns Trauma. 2014;2(1):24.
11. McGrath MM, Najafian N. The role of coinhibitory signaling pathways in transplantation and tolerance. Front Immunol. 2012;3.
12. Amouzegar A, Chauhan SK. Effector and Regulatory T Cell Trafficking in Corneal Allograft Rejection. Mediators Inflamm. 2017;2017:8670280.
13. Olson NC, Sallam R, Doyle MF, Tracy RP, Huber SA. T helper cell polarization in healthy people: implications for cardiovascular disease. J Cardiovasc Transl Res. 2013;6(5):772-86.
14. Trinchieri G. Interleukin-10 production by effector T cells: Th1 cells show self control. J Exp Med. 2007;204(2):239-43.
15. Linsley PS, Brady W, Urnes M, Grosmaire LS, Damle NK, Ledbetter JA. CTLA-4 is a second receptor for the B cell activation antigen B7. J Exp Med. 1991;174(3):561-9.
16. Chen L, Flies DB. Molecular mechanisms of T cell co-stimulation and co-inhibition. Nat Rev Immunol. 2013;13(4):227.
17. Pentcheva-Hoang T, Egen JG, Wojnoonski K, Allison JP. B7-1 and B7-2 selectively recruit CTLA-4 and CD28 to the immunological synapse. Immunity. 2004;21(3):401-13.
18. Liang SC, Latchman YE, Buhlmann JE, Tomczak MF, Horwitz BH, Freeman GJ, et al. Regulation of PD‐1, PD‐L1, and PD‐L2 expression during normal and autoimmune responses. Eur J Immunol. 2003;33(10):2706-16.
19. Gannavaram S, Bhattacharya P, Ismail N, Kaul A, Singh R, Nakhasi HL. Modulation of innate immune mechanisms to enhance Leishmania vaccine-induced immunity: role of coinhibitory molecules. Front Immunol. 2016;7:187.
20. Holmannová D, Kolackova M, Kondelkova K, Kunes P, Krejsek J, Andrys C. CD200/CD200R paired potent inhibitory molecules regulating immune and inflammatory responses; Part I: CD200/CD200R structure, activation, and function. acta Medica (hradec Králové). 2012;55(1):12-7.
21. Fakhari S, Kalantar E, Nikzaban M, Hakhamneshi MS, Fathi F, Nikkhoo B, et al. Effect of Helicobacter pylori infection on stromal-derived factor-1/CXCR4 axis in bone marrow-derived mesenchymal stem cells. Adv Biomed Res. 2014;3:19.
22. Domnina AP, Novikova PV, Lyublinskaya OG, Zenin VV, Fridlyanskaya II, Mikhailov VM, et al. Mesenchymal stem cells with irreversibly arrested proliferation stimulate decidua development in rats. Experimental and therapeutic medicine. 2016;12(4):2447-54.
23. Alipour R, Karimi MM, Hashemi-Beni B, Adib M, Sereshki N, Sadeghi F. Indoleamine 2, 3-dioxygenase is dispensable for the immunomodulatory function of stem cells from human exfoliated deciduous teeth. Cell J. 2017;18(4):597.
24. Chugh RM, Chaturvedi M, Yerneni LK. Exposure cell number during feeder cell growth-arrest by Mitomycin C is a critical pharmacological aspect in stem cell culture system. J Pharmacol Toxicol Methods. 2016;80:68-74.
25. Saldaña L, Bensiamar F, Vallés G, Mancebo FJ, García-Rey E, Vilaboa N. Immunoregulatory potential of mesenchymal stem cells following activation by macrophage-derived soluble factors. Stem Cell Res Ther. 2019;10(1):58.
26. Lim JY, Im KI, Lee ES, Kim N, Nam YS, Jeon YW, et al. Enhanced immunoregulation of mesenchymal stem cells by IL-10-producing type 1 regulatory T cells in collagen-induced arthritis. Sci Rep. 2016;6:26851.
27. Philipp D, Suhr L, Wahlers T, Choi YH, Paunel-Gorgulu A. Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization. Stem Cell Res Ther. 2018;9(1):286.
28. Hesami S, Mohammadi M, Rezaee MA, Jalili A, Rahmani MR. The effects of hyperthermia on the immunomodulatory properties of human umbilical cord vein mesenchymal stem cells (MSCs). Int J Hyperthermia. 2017;33(7):705-12.
29. Abolhasani M, Rezaee MA, Mohammadi M, Ghadimi T, Mohammadi M, Rahmani MR. Immunomodulatory properties of umbilical cord vein mesenchymal stromal cells influenced by gestational age and in vitro expansion. Immunol lett. 2018;194:62-8.
30. English K, Wood KJ. Mesenchymal stromal cells in transplantation rejection and tolerance. Cold Spring Harb Perspect Med. 2013;3(5):a015560.
31. Patsoukis N, Weaver JD, Strauss L, Herbel C, Seth P, Boussiotis VA. Immunometabolic regulations mediated by coinhibitory receptors and their impact on T cell immune responses. Front Immunol. 2017;8:330.
32. Fife BT, Bluestone JA. Control of peripheral T‐cell tolerance and autoimmunity via the CTLA‐4 and PD‐1 pathways. Immunol Rev. 2008;224(1):166-82.
33. Maccario R, Podestà M, Moretta A, Cometa A, Comoli P, Montagna D, et al. Interaction of human mesenchymal stem cells with cells involved in alloantigen-specific immune response favors the differentiation of CD4+ T-cell subsets expressing a regulatory/suppressive phenotype. Haematologica. 2005;90(4):516-25.
34. Yan Z, Zhuansun Y, Chen R, Li J, Ran P. Immunomodulation of mesenchymal stromal cells on regulatory T cells and its possible mechanism. Exp Cell Res. 2014;324(1):65-74.
35. Najar M, Raicevic G, Jebbawi F, De Bruyn C, Meuleman N, Bron D, et al. Characterization and functionality of the CD200–CD200R system during mesenchymal stromal cell interactions with T-lymphocytes. Immunol lett. 2012;146(1):50-6.
36. Wu Y, Hoogduijn MJ, Baan CC, Korevaar SS, Kuiper Rd, Yan L, et al. Adipose tissue-derived mesenchymal stem cells have a heterogenic cytokine secretion profile. Stem Cells Int. 2017;2017.
37. Liu Y, Beyer A, Aebersold R. On the dependency of cellular protein levels on mRNA abundance. Cell. 2016;165(3):535-50.
38. Parekkadan B, Milwid JM. Mesenchymal stem cells as therapeutics. Annu Rev Biomed Eng. 2010;12:87-117.
39. Najar M, Raicevic G, Fayyad-Kazan H, Bron D, Toungouz M, Lagneaux L. Mesenchymal stromal cells and immunomodulation: a gathering of regulatory immune cells. Cytotherapy. 2016;18(2):160-71.
40. Krampera M, Cosmi L, Angeli R, Pasini A, Liotta F, Andreini A, et al. Role for interferon‐γ in the immunomodulatory activity of human bone marrow mesenchymal stem cells. Stem cells. 2006;24(2):386-98.
41. Rocha PN, Plumb TJ, Crowley SD, Coffman TM. Effector mechanisms in transplant rejection. Immunol Rev. 2003;196(1):51-64.
42. Mojic M, Takeda K, Hayakawa Y. The Dark Side of IFN-γ: Its Role in Promoting Cancer Immunoevasion. Int J Mol Sci. 2017;19(1):89.
43. P De Miguel M, Fuentes-Julian S, Blazquez-Martinez A, Y Pascual C, A Aller M, Arias J, et al. Immunosuppressive properties of mesenchymal stem cells: advances and applications. Curr Mol Med. 2012;12(5):574-91.
44. Ivanova-Todorova E, Bochev I, Dimitrov R, Belemezova K, Mourdjeva M, Kyurkchiev S, et al. Conditioned Medium from Adipose Tissue-Derived Mesenchymal Stem Cells Induces CD4. J Biomed Biotechnol. 2012;2012.
45. Yang S, Li W, Liu W, Gao C, Zhou B, Li S, et al. IL-10 gene modified dendritic cells induced antigen-specific tolerance in experimental autoimmune myocarditis. Clin Immunol. 2006;121(1):63-73.
46. Jiang X-X, Zhang Y, Liu B, Zhang S-X, Wu Y, Yu X-D, et al. Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells. Blood. 2005;105(10):4120-6.
47. Keir ME, Butte MJ, Freeman GJ, Sharpe AH. PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol. 2008;26:677-704.
48. Del Rio ML, Buhler L, Gibbons C, Tian J, Rodriguez‐Barbosa JI. PD‐1/PD‐L1, PD‐1/PD‐L2, and other co‐inhibitory signaling pathways in transplantation. Transpl Int. 2008;21(11):1015-28.
49. Raimondi G, Shufesky WJ, Tokita D, Morelli AE, Thomson AW. Regulated compartmentalization of programmed cell death-1 discriminates CD4+ CD25+ resting regulatory T cells from activated T cells. J Immunol. 2006;176(5):2808-16.
50. Mimura K, Teh JL, Okayama H, Shiraishi K, Kua LF, Koh V, et al. PD‐L1 expression is mainly regulated by interferon gamma associated with JAK‐STAT pathway in gastric cancer. Cancer Sci. 2018.
51. Watanabe N, Nakajima H. Coinhibitory molecules in autoimmune diseases. Clin Dev Immunol. 2012;2012.
52. Ford ML. T cell cosignaling molecules in transplantation. Immunity. 2016;44(5):1020-33.
53. DeWolf S, Shen Y, Sykes M. A new window into the human alloresponse. Transplantation. 2016;100(8):1639-49.
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