Immune system-related plasma extracellular vesicles in healthy aging
ObjectivesTo identify age-related plasma extracellular vehicle (EVs) phenotypes in healthy adults.MethodsEV proteomics by high-resolution mass spectrometry to evaluate EV protein stability and discover age-associated EV proteins (n=4 with 4 serial freeze-thaws each); validation by high-resolution fl...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-04-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1355380/full |
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| author | Xin Zhang Xin Zhang Sisi Ma Janet L. Huebner Syeda Iffat Naz Noor Alnemer Erik J. Soderblom Constantin Aliferis Virginia Byers Kraus Virginia Byers Kraus Virginia Byers Kraus |
| author_facet | Xin Zhang Xin Zhang Sisi Ma Janet L. Huebner Syeda Iffat Naz Noor Alnemer Erik J. Soderblom Constantin Aliferis Virginia Byers Kraus Virginia Byers Kraus Virginia Byers Kraus |
| author_sort | Xin Zhang |
| collection | DOAJ |
| description | ObjectivesTo identify age-related plasma extracellular vehicle (EVs) phenotypes in healthy adults.MethodsEV proteomics by high-resolution mass spectrometry to evaluate EV protein stability and discover age-associated EV proteins (n=4 with 4 serial freeze-thaws each); validation by high-resolution flow cytometry and EV cytokine quantification by multiplex ELISA (n=28 healthy donors, aged 18-83 years); quantification of WI-38 fibroblast cell proliferation response to co-culture with PKH67-labeled young and old plasma EVs. The EV samples from these plasma specimens were previously characterized for bilayer structure, intra-vesicle mitochondria and cytokines, and hematopoietic cell-related surface markers.ResultsCompared with matched exo-EVs (EV-depleted supernatants), endo-EVs (EV-associated) had higher mean TNF-α and IL-27, lower mean IL-6, IL-11, IFN-γ, and IL-17A/F, and similar mean IL-1β, IL-21, and IL-22 concentrations. Some endo-EV and exo-EV cytokine concentrations were correlated, including TNF-α, IL-27, IL-6, IL-1β, and IFN-γ, but not IL-11, IL-17A/F, IL-21 or IL-22. Endo-EV IFN-γ and exo-EV IL-17A/F and IL-21 declined with age. By proteomics and confirmed by flow cytometry, we identified age-associated decline of fibrinogen (FGA, FGB and FGG) in EVs. Age-related EV proteins indicated predominant origins in the liver and innate immune system. WI-38 cells (>95%) internalized similar amounts of young and old plasma EVs, but cells that internalized PKH67-EVs, particularly young EVs, underwent significantly greater cell proliferation.ConclusionEndo-EV and exo-EV cytokines function as different biomarkers. The observed healthy aging EV phenotype reflected a downregulation of EV fibrinogen subpopulations consistent with the absence of a pro-coagulant and pro-inflammatory condition common with age-related disease. |
| first_indexed | 2024-04-24T14:26:38Z |
| format | Article |
| id | doaj.art-44b2324a23a2471499bb2ea7e95e68b3 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-24T14:26:38Z |
| publishDate | 2024-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-44b2324a23a2471499bb2ea7e95e68b32024-04-03T04:36:52ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-04-011510.3389/fimmu.2024.13553801355380Immune system-related plasma extracellular vesicles in healthy agingXin Zhang0Xin Zhang1Sisi Ma2Janet L. Huebner3Syeda Iffat Naz4Noor Alnemer5Erik J. Soderblom6Constantin Aliferis7Virginia Byers Kraus8Virginia Byers Kraus9Virginia Byers Kraus10Duke Molecular Physiology Institute, Duke University School of Medicine, Duke University, Durham, NC, United StatesDepartment of Orthopaedic Surgery, Duke University School of Medicine, Duke University, Durham, NC, United StatesInstitute for Health Informatics, University of Minnesota School of Medicine, Minneapolis, MN, United StatesDuke Molecular Physiology Institute, Duke University School of Medicine, Duke University, Durham, NC, United StatesInstitute for Health Informatics, University of Minnesota School of Medicine, Minneapolis, MN, United StatesDuke Molecular Physiology Institute, Duke University School of Medicine, Duke University, Durham, NC, United StatesDuke Proteomics and Metabolomics Core Facility, Duke University School of Medicine, Duke University, Durham, NC, United StatesInstitute for Health Informatics, University of Minnesota School of Medicine, Minneapolis, MN, United StatesDuke Molecular Physiology Institute, Duke University School of Medicine, Duke University, Durham, NC, United StatesDepartment of Orthopaedic Surgery, Duke University School of Medicine, Duke University, Durham, NC, United StatesDepartment of Medicine, Duke University School of Medicine, Duke University, Durham, NC, United StatesObjectivesTo identify age-related plasma extracellular vehicle (EVs) phenotypes in healthy adults.MethodsEV proteomics by high-resolution mass spectrometry to evaluate EV protein stability and discover age-associated EV proteins (n=4 with 4 serial freeze-thaws each); validation by high-resolution flow cytometry and EV cytokine quantification by multiplex ELISA (n=28 healthy donors, aged 18-83 years); quantification of WI-38 fibroblast cell proliferation response to co-culture with PKH67-labeled young and old plasma EVs. The EV samples from these plasma specimens were previously characterized for bilayer structure, intra-vesicle mitochondria and cytokines, and hematopoietic cell-related surface markers.ResultsCompared with matched exo-EVs (EV-depleted supernatants), endo-EVs (EV-associated) had higher mean TNF-α and IL-27, lower mean IL-6, IL-11, IFN-γ, and IL-17A/F, and similar mean IL-1β, IL-21, and IL-22 concentrations. Some endo-EV and exo-EV cytokine concentrations were correlated, including TNF-α, IL-27, IL-6, IL-1β, and IFN-γ, but not IL-11, IL-17A/F, IL-21 or IL-22. Endo-EV IFN-γ and exo-EV IL-17A/F and IL-21 declined with age. By proteomics and confirmed by flow cytometry, we identified age-associated decline of fibrinogen (FGA, FGB and FGG) in EVs. Age-related EV proteins indicated predominant origins in the liver and innate immune system. WI-38 cells (>95%) internalized similar amounts of young and old plasma EVs, but cells that internalized PKH67-EVs, particularly young EVs, underwent significantly greater cell proliferation.ConclusionEndo-EV and exo-EV cytokines function as different biomarkers. The observed healthy aging EV phenotype reflected a downregulation of EV fibrinogen subpopulations consistent with the absence of a pro-coagulant and pro-inflammatory condition common with age-related disease.https://www.frontiersin.org/articles/10.3389/fimmu.2024.1355380/fullextracellular vesicleshealthy agingimmune systemcytokinesproteomicssurface markers |
| spellingShingle | Xin Zhang Xin Zhang Sisi Ma Janet L. Huebner Syeda Iffat Naz Noor Alnemer Erik J. Soderblom Constantin Aliferis Virginia Byers Kraus Virginia Byers Kraus Virginia Byers Kraus Immune system-related plasma extracellular vesicles in healthy aging Frontiers in Immunology extracellular vesicles healthy aging immune system cytokines proteomics surface markers |
| title | Immune system-related plasma extracellular vesicles in healthy aging |
| title_full | Immune system-related plasma extracellular vesicles in healthy aging |
| title_fullStr | Immune system-related plasma extracellular vesicles in healthy aging |
| title_full_unstemmed | Immune system-related plasma extracellular vesicles in healthy aging |
| title_short | Immune system-related plasma extracellular vesicles in healthy aging |
| title_sort | immune system related plasma extracellular vesicles in healthy aging |
| topic | extracellular vesicles healthy aging immune system cytokines proteomics surface markers |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1355380/full |
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