MICROVESICLES DERIVED FROM MESENCHYMAL STEM CELLS HAVE ANTI-INFLAMMATORY AND IMMUNOMODULATORY EFFECTS ON PROINFLAMMATORY MACROPHAGES
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1
Federal University ofLaboratory of Clinical and Experimental Immunology, Department of Nephrology, Brazil
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2
University of Sao Paulo, Department of Immunology, Brazil
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3
University of Sao Paulo, Department of Pharmacology, Brazil
Introduction: The macrophages (Mɸs) are heterogeneous cells that contribute to renal regeneration and/or renal disease. M1 Mɸs secrete proinflammatory cytokines and reactive oxygen species (ROS) that enhance and exacerbate the inflammatory response to kidney injury. Conversely, M2 Mɸs release anti-inflammatory molecules and growth factors that promote renal regeneration and attenuate inflammation. Accordingly, the generation of regulatory Mɸs in the renal parenchyma turns into an important tool to establish alternative therapies in the treatment of acute and chronic renal disease. Interestingly, mesenchymal stem cells (MSCs) have a confirmed modulatory potential on various cells of the immune system. MSCs have been shown to be effective at educating Mɸs and reprogramming M1 into M2 Mɸs through paracrine secretion of TSG-6 and PGE2. It has been demonstrated that secretion of bioactive factors in the extracellular environment is the main mechanism by which occurs Mɸ modulation. In addition to this, it has recently been discovered that MSCs have the capacity to secrete renoprotective microvesicles with similar immunomodulatory functions. However, the physiological effect of MSCs-derived microvesicles (MSCs-MVs) on Mɸs remains unknown. Given that Mɸ modulation is a promising approach to renal protection, the aim of the study was to evaluate the immunoregulatory effect of MSCs-MVs on Mɸs. Methods: Mɸs derived from murine bone marrow prior converted to M1 phenotype (24h treatment with LPS/IFN-g) were co-cultured with MSCs (1:1 ratio) or MSC-MVs (dose of 50 μg every 8 hours) for 48 hours. Complementary, the effect of MSC-MVs on resident Mɸs was evaluated in vivo in a model of acute sterile peritonitis induced by intraperitoneal injection of 3% of thioglycollate in male C57BL/6 mice. The experiments were performed after characterization and polarization of Mɸs derived from bone marrow, differentiation of MSCs, MSC-MVs analysis and standardization of thioglycollate model. Results: Initially we verified and characterized the ability of Mɸs derived from bone marrow to be converted in M1 or M2 Mɸs. Later, we found that co-culture of MSCs with M1 Mɸs reduced the expression of M1 markers, such as CD86 and CCR7, and increased the expression of CD206 (M2). Additionally, treatment of M1 Mɸs with MSCs-MVs was effective in reducing the levels of pro-inflammatory molecules, such as CCR7, IL-1-β, IL-6 and NO, and was efficient in promoting a regulatory profile of treated Mɸs by increasing IL-10, CD206, and enzyme activity of arginase. We also studied the profile of miRNAs associated with Mɸ polarization in order to understand the mechanism involved in the Mɸ modulation. The expression of miR-155, a miR associated with the production of pro-inflammatory cytokines, was surprisingly reduced in Mɸs treated with MVs. SOCS3, one of miR-155 targets with a high immunoregulatory potencial, was increased in Mɸs exposed to MVs. This mechanism finally explains the reduced expression of proinflammatory molecules and the increased levels of M2 molecules in treated Mɸs. NanoSight analysis indicated that MVs-MSCs had an average size of 75 nm and expressed classical markers of MSCs on their surface, such as CD105, CD90, CD44 and CD73. The in vivo therapeutic potential of MSCs-MVs on resident Mɸs was confirmed in a model of acute sterile peritonitis. Primarily, we observed that the mice treated with 4 doses of MVs (100μg/24h) had a significant reduction in the number of recruited cells in the peritoneal cavity. Similarly, resident Mɸs (F4 /80 + CD11b +) of peritoneal lavage of these animals were in lower proportion than in control animals, and underexpressed CD86 and INOS. Interestingly, the treatment with MSCs-MVs increased the recruitment of myeloid cells defined as F4 /80 + CD11b- in the peritoneal cavity .These cells presented regulatory characteristics overexpressing CD206 and lacking expression of TLR4 and CD86. Finally, the anti-inflammatory microenvironment generated by MSCs-MVS was confirmed by the reduced levels of IFN-g, IL-b and TNF-a in the peritoneal cavity of treated animals. Conclusion: Our results suggest that the MSCs-MVs are both effective in vitro and in vivo in modulating and converting M1 Mɸs into a more regulatory profile (MSCs-MVs- derived M1), This modulation was confirmed in the model of acute sterile peritonitis induced by thioglycollate. MVs were able to reduce the number of resident Mɸs and also the expression of M1 markers in the treated animals. Additionally, we believe that the mechanism by which the modulation occurs may be associated with a reduction of miR-155 levels, which leads to increased expression of SOCS3 and decreased levels of proinflammatory cytokines and increase of M2 markers expression. This result opens a new way to explore the MSCs-MVs in various inflammatory diseases characterized by an augmentation of M1 Mɸs, such as diabetes, arthritis and kidney disease.
Acknowledgements
Acknowledgments to government funding agencies:
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Ministério da Educação do brasil (MEC)
References
Park CW. Diabetic kidney disease: from epidemiology to clinical perspectives. Diabetes Metab J. 2014;38(4):252-60.
de Almeida DC, Donizetti-Oliveira C, Barbosa-Costa P, Origassa CS, Camara NO. In search of mechanisms associated with mesenchymal stem cell-based therapies for acute kidney injury. The Clinical biochemist Reviews / Australian Association of Clinical Biochemists. 2013;34(3):131-44.
Ricardo SD, van Goor H, Eddy AA. Macrophage diversity in renal injury and repair. The Journal of clinical investigation. 2008;118(11):3522-30.
Nombela-Arrieta C, Ritz J, Silberstein LE. The elusive nature and function of mesenchymal stem cells. Nature reviews Molecular cell biology. 2011;12(2):126-31.
Murphy MB, Moncivais K, Caplan AI. Mesenchymal stem cells: environmentally responsive therapeutics for regenerative medicine. Experimental & molecular medicine. 2013;45:e54.
Martinez FO, Gordon S, Locati M, Mantovani A. Transcriptional profiling of the human monocyte-to-macrophage differentiation and polarization: new molecules and patterns of gene expression. Journal of immunology. 2006;177(10):7303-11.
Biancone L, Bruno S, Deregibus MC, Tetta C, Camussi G. Therapeutic potential of mesenchymal stem cell-derived microvesicles. Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association. 2012;27(8):3037-42.
Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M. The chemokine system in diverse forms of macrophage activation and polarization. Trends in immunology. 2004;25(12):677-86.
Keywords:
Mesenchymal Stem Cells,
Microvesicles,
Immune Regulation,
Macrophages,
polarization
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Oral Presentation
Topic:
Innate Immunity
Citation:
Henao
JS,
Braga
TT,
Amano
MT,
Cenedeze
MA,
Pereira
RL,
Muscará
MN,
Teixeira
SA,
Watanabe
IK,
Pacheco-Silva
A,
Almeida
DC and
Câmara
NO
(2015). MICROVESICLES DERIVED FROM MESENCHYMAL STEM CELLS HAVE ANTI-INFLAMMATORY AND IMMUNOMODULATORY EFFECTS ON PROINFLAMMATORY MACROPHAGES.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00282
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Received:
14 May 2015;
Published Online:
15 Sep 2015.
*
Correspondence:
Mr. Juan S Henao, Federal University ofLaboratory of Clinical and Experimental Immunology, Department of Nephrology, Laboratory of Clinical and Experimental Immunology, Brazil, juanelmono17@hotmail.com