Whole platelets induces macrophage programming toward a repair profile characterized by high IL-10 production
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1
Universidad de Chile, Facultad de Medicina, Chile
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2
Hospital Clínico de la Universidad de Chile, Chile
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3
Universidad de Chile, Facultad de Odontología, Instituto de Investigación de Ciencias Odontológicas, Chile
Introduction:
Macrophages are highly plastic cells which when activated can polarize toward two established profiles. The conventional polarization generates proinflammatory macrophage and is associated with acute inflammation, the alternative activation, on the contrary, generates macrophages involved in tissue repair. Both types of macrophages are related to many pathological conditions including inflammation, vascular disorders and cancer. Furthermore, platelets are cellular fragments involved in processes of inflammation and tissue regeneration. Their activation releases several molecules that promote differentiation of blood monocytes into non-polarized macrophages. However, it is unclear if platelets play a role in the subsequent activation process that can generate macrophages with an inflammatory or repair profile. Understanding the mechanisms by which platelets and macrophages interact, two key players, for example, of tumor progression, would allow us to modulate the tumoral niche. This study focuses in the effect of platelets on non-polarized macrophages. In this regard we propose that platelets are able to polarize and re-polarize macrophages toward a repair profile.
Materials and Methods:
From healthy donors of Blood Bank, it was obtained Platelet Rich Plasma (PRP) and Plasma Rich in Growth Factors (PRGF) (PRP-derivated from the activation with calcium chloride). In PRP we measured the platelet concentration, by hemocytometry, and its purity, by flow cytometry analysis of CD41-labeled platelets. PRGF was characterized by TGF-β, IL-1β, TNF-α and IL-10 cytokine concentration by ELISA. Simultaneously, CD14+ monocytes purified by "Rosettes" from fresh buffy coat were cultured in RPMI / SFB / M-CSF for seven days. Then, cells were harvested and characterized morphologically and phenotypically by light microscopy and flow cytometry analysis of CD32, CD11c and CD14 (monocyte-macrophage lineage associated markers), respectively. The platelet-macrophage interaction was tested using SRFC or whole platelets (PRP) to stimulate macrophages for 24 h. A phenotypic characterization was examined by measuring costimulatory molecule CD86 and markers associated repair profile CD163 and CD206 (flow cytometry). IL-10 and TNF-α cytokine production in the supernatant of these macrophages was measured by ELISA. Additionally, proinflammatory macrophage, obtained by stimulating macrophages with LPS and IFN-γ for 24 h., were co-incubated with PRP or PRGF for 24 h. and their re-polarization was examined by flow cytometry analysis of CD206, CD86 and CD163.
Results:
The human monocyte derivated macrophages presented a typical morphology and expressed the classic markers of macrophage-monocyte, i.e. CD11c, CD14 and CD32. Stimulation of these non-polarized macrophages with SRFC, which has a high concentration of TGF-β, did not generate statistically significant differences in the expression of CD206, CD163 and CD86 markers, neither in the cytokine production, compared to unstimulated macrophage. In contrast, upon stimulation of non-polarized and proinflammatory macrophages with PRP (whole platelets) we found an increase in the CD206 mannose receptor expression and in the IL-10 levels compared to the unstimulated control. Both IL-10 and CD206 are characteristic of the repairing or alternative activated profile.
Discussion:
In this project it was demonstrated in vitro that whole platelets induce macrophage polarization toward a repair phenotype at 24 h., characterized by high levels of mannose receptor expression and the production of the potent anti-inflammatory cytokine IL-10. Furthermore, platelets might be able to re-polarize proinflammatory macrophage into a repair phenotype. Further investigation allow to validate these results.
This interaction could occur in vivo, so it is necessary to consider this communications between platelets and macrophages in the inflammation or tissue repair an injury, together with their potential use for the development of therapeutic strategies to treat diseases associated with chronic inflammation. Finally, this project is a first attempt to establish whether there is a communication or "cross-talk" between this two actors that could participate in tumor niche, promoting a tolerant microenvironment that encourages the tumor maintenance and spread.
Acknowledgements
Acknowledgements:
- Fundación Ciencia Translacional, ICM funds project N° P. IMII P09/016-F.
- Fondecyt project N°1130324.
Keywords:
Macrophages,
tissue repair,
platelets,
polarization,
Mannose receptor,
tumor progression,
Anti-Inflammatory Agents,
M2 macrophages
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:
Poster Presentation
Topic:
Innate Immunity
Citation:
Escobar Arrepol
GS,
Pérez
C,
Escobar
A,
Ortiz
MC,
Tempio Sepúlveda
F and
Lopez
MN
(2015). Whole platelets induces macrophage programming toward a repair profile characterized by high IL-10 production.
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.2015.05.00015
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Received:
15 May 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
Miss. Gisselle S Escobar Arrepol, Universidad de Chile, Facultad de Medicina, Santiago, Chile, gescobar@ug.uchile.cl