Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner
ObjectiveIn a recent study, we found an elevated level of interleukin 8 (IL-8) in response to bacterial incubation in thrombin-sufficient human whole blood anticoagulated by the fibrin polymerization blocking peptide GPRP. Whether thrombin directly activated leukocytes or mediated the release via th...
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| Format: | Article |
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.865386/full |
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| author | Huy Quang Quach Christina Johnson Karin Ekholt Rakibul Islam Tom Eirik Mollnes Tom Eirik Mollnes Tom Eirik Mollnes Per H. Nilsson Per H. Nilsson |
| author_facet | Huy Quang Quach Christina Johnson Karin Ekholt Rakibul Islam Tom Eirik Mollnes Tom Eirik Mollnes Tom Eirik Mollnes Per H. Nilsson Per H. Nilsson |
| author_sort | Huy Quang Quach |
| collection | DOAJ |
| description | ObjectiveIn a recent study, we found an elevated level of interleukin 8 (IL-8) in response to bacterial incubation in thrombin-sufficient human whole blood anticoagulated by the fibrin polymerization blocking peptide GPRP. Whether thrombin directly activated leukocytes or mediated the release via thrombin-dependent activation of platelets remains unresolved. Herein, we addressed the role of thrombin and platelets in IL-8 release.MethodsWe separated platelets from whole blood using a combination of 0.7% (w/v) citrate and GPRP for attenuating the hemostatic response during the separation of platelets. Cytokine responses were compared in whole blood and platelet-depleted blood upon Escherichia coli incubation. Cytokine responses were also profiled with and without reconstitution of either platelets or the supernatant from activated platelets.ResultsPlatelets were not activated during the separation process but responded to stimuli upon re-calcification. Plasma levels of IL-1β, IL-1Ra, IL-6, IL-8, IP-10, MIP-1α, and MIP-1β were significantly reduced in platelet-depleted blood compared to whole blood, but recovered in the presence of platelets, or with the supernatant of activated platelets. The leukocyte fraction and platelets were each found to contribute to the elevation of IL-8 at around 5 ng/ml; however, if combined, the release of IL-8 increased to 26 ng/ml. This process was dependent on thrombin since the levels of IL-8 remained at 5 ng/ml in whole blood if thrombin was blocked. Intracellular staining revealed that monocytes were the main source for IL-8 expression.ConclusionOur findings suggest that the release of IL-8 is mediated by the leukocytes, mainly monocytes, but potentiated via thrombin-dependent activation of platelets. |
| first_indexed | 2024-12-13T19:32:02Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-13T19:32:02Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-56808b621439489dbbf559a55ea9a72d2022-12-21T23:33:54ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-04-011310.3389/fimmu.2022.865386865386Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent MannerHuy Quang Quach0Christina Johnson1Karin Ekholt2Rakibul Islam3Tom Eirik Mollnes4Tom Eirik Mollnes5Tom Eirik Mollnes6Per H. Nilsson7Per H. Nilsson8Department of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayDepartment of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayDepartment of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayDepartment of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayDepartment of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayResearch Laboratory, Nordland Hospital, K.G Jebsen Center TREC, University of Tromsø, Bodø, NorwayCentre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, NorwayDepartment of Immunology, Oslo University Hospital Rikshospitalet, University of Oslo, Oslo, NorwayDepartment of Chemistry and Biomedicine, Linnaeus Centre for Biomaterials Chemistry, Linnaeus University, Kalmar, SwedenObjectiveIn a recent study, we found an elevated level of interleukin 8 (IL-8) in response to bacterial incubation in thrombin-sufficient human whole blood anticoagulated by the fibrin polymerization blocking peptide GPRP. Whether thrombin directly activated leukocytes or mediated the release via thrombin-dependent activation of platelets remains unresolved. Herein, we addressed the role of thrombin and platelets in IL-8 release.MethodsWe separated platelets from whole blood using a combination of 0.7% (w/v) citrate and GPRP for attenuating the hemostatic response during the separation of platelets. Cytokine responses were compared in whole blood and platelet-depleted blood upon Escherichia coli incubation. Cytokine responses were also profiled with and without reconstitution of either platelets or the supernatant from activated platelets.ResultsPlatelets were not activated during the separation process but responded to stimuli upon re-calcification. Plasma levels of IL-1β, IL-1Ra, IL-6, IL-8, IP-10, MIP-1α, and MIP-1β were significantly reduced in platelet-depleted blood compared to whole blood, but recovered in the presence of platelets, or with the supernatant of activated platelets. The leukocyte fraction and platelets were each found to contribute to the elevation of IL-8 at around 5 ng/ml; however, if combined, the release of IL-8 increased to 26 ng/ml. This process was dependent on thrombin since the levels of IL-8 remained at 5 ng/ml in whole blood if thrombin was blocked. Intracellular staining revealed that monocytes were the main source for IL-8 expression.ConclusionOur findings suggest that the release of IL-8 is mediated by the leukocytes, mainly monocytes, but potentiated via thrombin-dependent activation of platelets.https://www.frontiersin.org/articles/10.3389/fimmu.2022.865386/fullplateletsIL-8acute inflammationhuman whole bloodthrombincytokines |
| spellingShingle | Huy Quang Quach Christina Johnson Karin Ekholt Rakibul Islam Tom Eirik Mollnes Tom Eirik Mollnes Tom Eirik Mollnes Per H. Nilsson Per H. Nilsson Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner Frontiers in Immunology platelets IL-8 acute inflammation human whole blood thrombin cytokines |
| title | Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner |
| title_full | Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner |
| title_fullStr | Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner |
| title_full_unstemmed | Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner |
| title_short | Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner |
| title_sort | platelet depletion of whole blood reveals that platelets potentiate the release of il 8 from leukocytes into plasma in a thrombin dependent manner |
| topic | platelets IL-8 acute inflammation human whole blood thrombin cytokines |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.865386/full |
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