Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals
In kidney stone disease, macrophages secrete various mediators via classical secretory pathway and cause renal interstitial inflammation. However, whether their extracellular vesicles, particularly exosomes, are involved in kidney stone pathogenesis remained unknown. This study investigated alterati...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-02-01
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| Series: | Frontiers in Immunology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fimmu.2018.00316/full |
| _version_ | 1819161795939532800 |
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| author | Nilubon Singhto Nilubon Singhto Rattiyaporn Kanlaya Rattiyaporn Kanlaya Angkhana Nilnumkhum Angkhana Nilnumkhum Visith Thongboonkerd Visith Thongboonkerd |
| author_facet | Nilubon Singhto Nilubon Singhto Rattiyaporn Kanlaya Rattiyaporn Kanlaya Angkhana Nilnumkhum Angkhana Nilnumkhum Visith Thongboonkerd Visith Thongboonkerd |
| author_sort | Nilubon Singhto |
| collection | DOAJ |
| description | In kidney stone disease, macrophages secrete various mediators via classical secretory pathway and cause renal interstitial inflammation. However, whether their extracellular vesicles, particularly exosomes, are involved in kidney stone pathogenesis remained unknown. This study investigated alterations in exosomal proteome of U937-derived macrophages (by phorbol-12-myristate-13-acetate activation) after exposure to calcium oxalate monohydrate (COM) crystals for 16-h using 2-DE-based proteomics approach. Six significantly altered proteins in COM-treated exosomes were successfully identified by nanoscale liquid chromatography–electrospray ionization–electron transfer dissociation tandem mass spectrometry as proteins involved mainly in immune processes, including T-cell activation and homeostasis, Fcγ receptor-mediated phagocytosis, interferon-γ (IFN-γ) regulation, and cell migration/movement. The decreased heat shock protein 90-beta (HSP90β) and increased vimentin were confirmed by Western blotting. ELISA showed that the COM-treated macrophages produced greater level of interleukin-1β (IL-1β), one of the markers for inflammasome activation. Functional studies demonstrated that COM-treated exosomes enhanced monocyte and T-cell migration, monocyte activation and macrophage phagocytic activity, but on the other hand, reduced T-cell activation. In addition, COM-treated exosomes enhanced production of proinflammatory cytokine IL-8 by monocytes that could be restored to its basal level by small-interfering RNA targeting on vimentin (si-Vimentin). Moreover, si-Vimentin could also abolish effects of COM-treated exosomes on monocyte and T-cell migration as well as macrophage phagocytic activity. These findings provided some implications to the immune response during kidney stone pathogenesis via exosomal pathway of macrophages after exposure to COM crystals. |
| first_indexed | 2024-12-22T17:18:02Z |
| format | Article |
| id | doaj.art-fa5b0257d1644240a75446e8d2cf21a9 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-22T17:18:02Z |
| publishDate | 2018-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-fa5b0257d1644240a75446e8d2cf21a92022-12-21T18:18:55ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-02-01910.3389/fimmu.2018.00316329563Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate CrystalsNilubon Singhto0Nilubon Singhto1Rattiyaporn Kanlaya2Rattiyaporn Kanlaya3Angkhana Nilnumkhum4Angkhana Nilnumkhum5Visith Thongboonkerd6Visith Thongboonkerd7Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandImmunology Graduate Program, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandMedical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandCenter for Research in Complex Systems Science, Mahidol University, Bangkok, ThailandMedical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandCenter for Research in Complex Systems Science, Mahidol University, Bangkok, ThailandMedical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandCenter for Research in Complex Systems Science, Mahidol University, Bangkok, ThailandIn kidney stone disease, macrophages secrete various mediators via classical secretory pathway and cause renal interstitial inflammation. However, whether their extracellular vesicles, particularly exosomes, are involved in kidney stone pathogenesis remained unknown. This study investigated alterations in exosomal proteome of U937-derived macrophages (by phorbol-12-myristate-13-acetate activation) after exposure to calcium oxalate monohydrate (COM) crystals for 16-h using 2-DE-based proteomics approach. Six significantly altered proteins in COM-treated exosomes were successfully identified by nanoscale liquid chromatography–electrospray ionization–electron transfer dissociation tandem mass spectrometry as proteins involved mainly in immune processes, including T-cell activation and homeostasis, Fcγ receptor-mediated phagocytosis, interferon-γ (IFN-γ) regulation, and cell migration/movement. The decreased heat shock protein 90-beta (HSP90β) and increased vimentin were confirmed by Western blotting. ELISA showed that the COM-treated macrophages produced greater level of interleukin-1β (IL-1β), one of the markers for inflammasome activation. Functional studies demonstrated that COM-treated exosomes enhanced monocyte and T-cell migration, monocyte activation and macrophage phagocytic activity, but on the other hand, reduced T-cell activation. In addition, COM-treated exosomes enhanced production of proinflammatory cytokine IL-8 by monocytes that could be restored to its basal level by small-interfering RNA targeting on vimentin (si-Vimentin). Moreover, si-Vimentin could also abolish effects of COM-treated exosomes on monocyte and T-cell migration as well as macrophage phagocytic activity. These findings provided some implications to the immune response during kidney stone pathogenesis via exosomal pathway of macrophages after exposure to COM crystals.http://journal.frontiersin.org/article/10.3389/fimmu.2018.00316/fullcalcium oxalatecalcium oxalate monohydrateinflammasomeinflammationkidney stonemigration |
| spellingShingle | Nilubon Singhto Nilubon Singhto Rattiyaporn Kanlaya Rattiyaporn Kanlaya Angkhana Nilnumkhum Angkhana Nilnumkhum Visith Thongboonkerd Visith Thongboonkerd Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals Frontiers in Immunology calcium oxalate calcium oxalate monohydrate inflammasome inflammation kidney stone migration |
| title | Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals |
| title_full | Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals |
| title_fullStr | Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals |
| title_full_unstemmed | Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals |
| title_short | Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals |
| title_sort | roles of macrophage exosomes in immune response to calcium oxalate monohydrate crystals |
| topic | calcium oxalate calcium oxalate monohydrate inflammasome inflammation kidney stone migration |
| url | http://journal.frontiersin.org/article/10.3389/fimmu.2018.00316/full |
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