Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity
Background: The liver-derived plasma protein fetuin-A is a systemic inhibitor of ectopic calcification. Fetuin-A stabilizes saturated mineral solutions by forming colloidal protein-mineral complexes called calciprotein particles (CPP). CPP are initially spherical, amorphous and soft, and are referre...
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| Format: | Article |
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Frontiers Media S.A.
2018-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2018.01991/full |
| _version_ | 1811279481155878912 |
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| author | Sina Köppert Andrea Büscher Anne Babler Ahmed Ghallab Ahmed Ghallab Eva M. Buhl Eicke Latz Jan G. Hengstler Edward R. Smith Edward R. Smith Willi Jahnen-Dechent |
| author_facet | Sina Köppert Andrea Büscher Anne Babler Ahmed Ghallab Ahmed Ghallab Eva M. Buhl Eicke Latz Jan G. Hengstler Edward R. Smith Edward R. Smith Willi Jahnen-Dechent |
| author_sort | Sina Köppert |
| collection | DOAJ |
| description | Background: The liver-derived plasma protein fetuin-A is a systemic inhibitor of ectopic calcification. Fetuin-A stabilizes saturated mineral solutions by forming colloidal protein-mineral complexes called calciprotein particles (CPP). CPP are initially spherical, amorphous and soft, and are referred to as primary CPP. These particles spontaneously convert into secondary CPP, which are larger, oblongate, more crystalline, and less soluble. CPP mediate excess mineral transport and clearance from circulation.Methods: We studied by intravital two-photon microscopy the clearance of primary vs. secondary CPP by injecting i.v. synthetic fluorescent CPP in mice. We analyzed CPP organ distribution and identified CPP endocytosing cells by immunofluorescence. Cellular clearance was studied using bone marrow-derived mouse wildtype and scavenger receptor A (SRA)-deficient macrophages, as well as human umbilical cord endothelial cells (HUVEC), monocyte-derived macrophages (hMDM), and human aortic endothelial cells (haEC). We employed mouse wildtype and mutant immortalized macrophages to analyze CPP-induced inflammasome activation and cytokine secretion.Results: In live mice, only primary CPP were rapidly cleared by liver sinusoidal endothelial cells (LSEC), whereas primary and secondary CPP were cleared by Kupffer cells. Scavenger receptor A (SRA)-deficient bone marrow macrophages endocytosed secondary CPP less well than did wildtype macrophages. In contrast, primary CPP endocytosis did not depend on the presence of SRA, suggesting involvement of an alternative clearance pathway. CPP triggered TLR4 dependent TNFα and IL-1β secretion in cultured macrophages. Calcium content-matched primary CPP caused twice more IL-1β secretion than did secondary CPP, which was associated with increased calcium-dependent inflammasome activation, suggesting that intracellular CPP dissolution and calcium overload may cause this inflammation.Conclusions: Secondary CPP are endocytosed by macrophages in liver and spleen via SRA. In contrast, our results suggest that primary CPP are cleared by LSEC via an alternative pathway. CPP induced TLR4-dependent TNFα and inflammasome-dependent IL-1β secretion in macrophages suggesting that inflammation and calcification may be considered consequences of prolonged CPP presence and clearance. |
| first_indexed | 2024-04-13T00:55:23Z |
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| language | English |
| last_indexed | 2024-04-13T00:55:23Z |
| publishDate | 2018-09-01 |
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| spelling | doaj.art-7c877a43aa93416580c00deb29a92fa52022-12-22T03:09:40ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-09-01910.3389/fimmu.2018.01991395367Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and CrystallinitySina Köppert0Andrea Büscher1Anne Babler2Ahmed Ghallab3Ahmed Ghallab4Eva M. Buhl5Eicke Latz6Jan G. Hengstler7Edward R. Smith8Edward R. Smith9Willi Jahnen-Dechent10Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Hospital, Aachen, GermanyHelmholtz-Institute for Biomedical Engineering, RWTH Aachen University Hospital, Aachen, GermanyHelmholtz-Institute for Biomedical Engineering, RWTH Aachen University Hospital, Aachen, GermanyLeibniz Research Centre for Working Environment and Human Factors, >Dortmund, GermanyDepartment of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, Qena, EgyptElectron Microscopy Facility, RWTH Aachen University Hospital, Aachen, GermanyInstitute of Innate Immunity, University Hospital Bonn, Bonn, GermanyLeibniz Research Centre for Working Environment and Human Factors, >Dortmund, GermanyDepartment of Nephrology, The Royal Melbourne Hospital, Melbourne, VIC, AustraliaDepartment of Medicine, University of Melbourne, Parkville, VIC, AustraliaHelmholtz-Institute for Biomedical Engineering, RWTH Aachen University Hospital, Aachen, GermanyBackground: The liver-derived plasma protein fetuin-A is a systemic inhibitor of ectopic calcification. Fetuin-A stabilizes saturated mineral solutions by forming colloidal protein-mineral complexes called calciprotein particles (CPP). CPP are initially spherical, amorphous and soft, and are referred to as primary CPP. These particles spontaneously convert into secondary CPP, which are larger, oblongate, more crystalline, and less soluble. CPP mediate excess mineral transport and clearance from circulation.Methods: We studied by intravital two-photon microscopy the clearance of primary vs. secondary CPP by injecting i.v. synthetic fluorescent CPP in mice. We analyzed CPP organ distribution and identified CPP endocytosing cells by immunofluorescence. Cellular clearance was studied using bone marrow-derived mouse wildtype and scavenger receptor A (SRA)-deficient macrophages, as well as human umbilical cord endothelial cells (HUVEC), monocyte-derived macrophages (hMDM), and human aortic endothelial cells (haEC). We employed mouse wildtype and mutant immortalized macrophages to analyze CPP-induced inflammasome activation and cytokine secretion.Results: In live mice, only primary CPP were rapidly cleared by liver sinusoidal endothelial cells (LSEC), whereas primary and secondary CPP were cleared by Kupffer cells. Scavenger receptor A (SRA)-deficient bone marrow macrophages endocytosed secondary CPP less well than did wildtype macrophages. In contrast, primary CPP endocytosis did not depend on the presence of SRA, suggesting involvement of an alternative clearance pathway. CPP triggered TLR4 dependent TNFα and IL-1β secretion in cultured macrophages. Calcium content-matched primary CPP caused twice more IL-1β secretion than did secondary CPP, which was associated with increased calcium-dependent inflammasome activation, suggesting that intracellular CPP dissolution and calcium overload may cause this inflammation.Conclusions: Secondary CPP are endocytosed by macrophages in liver and spleen via SRA. In contrast, our results suggest that primary CPP are cleared by LSEC via an alternative pathway. CPP induced TLR4-dependent TNFα and inflammasome-dependent IL-1β secretion in macrophages suggesting that inflammation and calcification may be considered consequences of prolonged CPP presence and clearance.https://www.frontiersin.org/article/10.3389/fimmu.2018.01991/fullcalciprotein particlecalcificationinflammationphosphatefetuin-Aplasma protein |
| spellingShingle | Sina Köppert Andrea Büscher Anne Babler Ahmed Ghallab Ahmed Ghallab Eva M. Buhl Eicke Latz Jan G. Hengstler Edward R. Smith Edward R. Smith Willi Jahnen-Dechent Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity Frontiers in Immunology calciprotein particle calcification inflammation phosphate fetuin-A plasma protein |
| title | Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity |
| title_full | Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity |
| title_fullStr | Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity |
| title_full_unstemmed | Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity |
| title_short | Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity |
| title_sort | cellular clearance and biological activity of calciprotein particles depend on their maturation state and crystallinity |
| topic | calciprotein particle calcification inflammation phosphate fetuin-A plasma protein |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2018.01991/full |
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