{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T12:20:10Z","timestamp":1790252410944,"version":"4.1.0"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,1,3]],"date-time":"2024-01-03T00:00:00Z","timestamp":1704240000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Thrombocytopenia is the most notable phenomenon in dengue. Activation status of platelets and interaction of platelets with endothelium contribute towards dengue disease pathogenesis. Platelets are the major cell types known to release extracellular vesicles, especially exosomes in circulation. However, the role of platelet derived exosomes (PLT-EXOs) in endothelial dysfunction during dengue infection remains unknown.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study, we recruited 28 healthy subjects and 69 dengue patients categorized as WS- (n=31), WS+ (n=29) and SD (n=9). Platelets were isolated from platelet rich plasma of dengue patients and their activation was assessed by flow cytometry. PLT-EXOs were isolated by ultracentrifugation method. Western blot analyses were performed to characterize the exosomes. Exosome uptake experiment was carried out to see the internalization of exosomes inside endothelial cells (HUVECs). To observe the effect of exosomes on endothelial cells, exosomes were added on HUVECs and expression of adherens and tight junctional proteins were examined by immunofluorescence assay and western blot. Expression levels of vascular injury markers were measured in the culture supernatants of Exosome-HUVEC coculture and sera of dengue patients by MSD-multiplex assay.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>As compared to healthy subjects, CD41\/CD61 expression was significantly reduced (p&amp;lt;0.0001) and CD62p expression was significantly increased (p&amp;lt;0.0001) on platelets in dengue patients. PLT-EXOs isolated from the dengue patients showed higher expression of CD63 and CD9 proteins than the healthy subjects. With <jats:italic>in-vitro<\/jats:italic> immunofluorescence assays, we illustrated the internalization of PLT-EXOs by the HUVECs and observed disruption of endothelial cell monolayer integrity in the presence of PLT-EXOs from WS+ and SD patients. Furthermore, the significant reduction in the expressions of ZO-2, VE-Cadherin and CD31 in endothelial cells following exposure to PLT-EXOs from the dengue patients provide direct evidence of PLT-EXOs mediated vascular permeability. PLT-EXOs stimulated the release of inflammatory markers CRP, SAA, sVCAM-1 and sICAM-1 in the supernatants of HUVEC cells. Importantly, significantly higher levels of CRP, sVCAM-1 and sICAM-1 in the sera of severe than mild dengue patients (p&amp;lt;0.0001) suggest their role in disease severity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>In summary, our data suggest that PLT-EXOs promote vascular leakage via release of proinflammatory mediators and compromise vascular barrier integrity in dengue patients.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1285162","type":"journal-article","created":{"date-parts":[[2024,1,3]],"date-time":"2024-01-03T04:31:57Z","timestamp":1704256317000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":30,"title":["Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients"],"prefix":"10.3389","volume":"14","author":[{"given":"Sayali","family":"Vedpathak","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Archana","family":"Sharma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonali","family":"Palkar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Varsha R.","family":"Bhatt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vishwanath Chandrashekhar","family":"Patil","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arjun L.","family":"Kakrani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"AkhileshChandra","family":"Mishra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepak","family":"Bhosle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vidya A.","family":"Arankalle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shubham","family":"Shrivastava","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,1,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-019-0476-8","article-title":"The current and future global distribution and population at risk of dengue","volume":"4","author":"Messina","year":"2019","journal-title":"Nat Microbiol"},{"key":"B2","volume-title":"Dengue: Guidelines for diagnosis, treatment, prevention and control","year":"2009"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001397","article-title":"Evaluation of the traditional and revised WHO classifications of Dengue disease severity","volume":"5","author":"Narvaez","year":"2011","journal-title":"PloS Negl Trop Dis"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3003084","article-title":"Dynamics of dengue disease severity determined by the interplay between viral genetics and serotype-specific immunity","volume":"3","author":"OhAinle","year":"2011","journal-title":"Sci Transl Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1111\/imm.12748","article-title":"Pathogenesis of vascular leak in dengue virus infection","volume":"151","author":"Malavige","year":"2017","journal-title":"Immunology"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.coviro.2020.06.008","article-title":"Risk factors and biomarkers of severe dengue","volume":"43","author":"Rathore","year":"2020","journal-title":"Curr Opin Virol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cix567","article-title":"Endothelial nitric oxide pathways in the pathophysiology of dengue: A prospective observational study","volume":"65","author":"Yacoub","year":"2017","journal-title":"Clin Infect Dis"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.aaa3863","article-title":"Dengue virus NS1 protein activates cells via Toll-like receptor 4 and disrupts endothelial cell monolayer integrity","volume":"7","author":"Modhiran","year":"2015","journal-title":"Sci Transl Med"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006673","article-title":"Dengue virus NS1 cytokine-independent vascular leak is dependent on endothelial glycocalyx components","volume":"13","author":"Glasner","year":"2017","journal-title":"PloS Pathog"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/srep46191","article-title":"Increased serum hyaluronic acid and heparan sulfate in dengue fever: association with plasma leakage and disease severity","volume":"7","author":"Tang","year":"2017","journal-title":"Sci Rep"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1400091","article-title":"Platelet activation and apoptosis modulate monocyte inflammatory responses in dengue","volume":"193","author":"Hottz","year":"2014","journal-title":"J Immunol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.7883\/yoken.JJID.2016.123","article-title":"Synergistic effect of TNF-\u03b1 and dengue virus infection on adhesion molecule reorganization in human endothelial cells","volume":"70","author":"Inyoo","year":"2017","journal-title":"Jpn J Infect Dis"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2013-05-504449","article-title":"Platelets mediate increased endothelium permeability in dengue through NLRP3-inflammasome activation","volume":"122","author":"Hottz","year":"2013","journal-title":"Blood"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/srep41697","article-title":"Platelet activation determines the severity of thrombocytopenia in dengue infection","volume":"7","author":"Ojha","year":"2017","journal-title":"Sci Rep"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/313842","article-title":"Thrombocytopenia in dengue: interrelationship between virus and the imbalance between coagulation and fibrinolysis and inflammatory mediators","volume":"2015","author":"de Azeredo","year":"2015","journal-title":"Mediators Inflamm"},{"key":"B16","article-title":"Dengue virus and endothelial cell: a related phenomenon to thrombocytopenia and granulocytopenia in dengue hemorrhagic fever","author":"Butthep","year":"1993","journal-title":"Southeast Asian J Trop Med Public Health"},{"key":"B17","doi-asserted-by":"publisher","first-page":"2402","DOI":"10.1038\/s41467-019-10360-4","article-title":"Extracellular vesicles from CLEC2-activated platelets enhance dengue virus-induced lethality via CLEC5A\/TLR2","volume":"10","author":"Sung","year":"2019","journal-title":"Nat Commun"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003459","article-title":"Platelet activating factor contributes to vascular leak in acute dengue infection","volume":"9","author":"Jeewandara","year":"2015","journal-title":"PloS Negl Trop Dis"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.616394","article-title":"Exposure of platelets to dengue virus and envelope protein domain III induces nlrp3 inflammasome-dependent platelet cell death and thrombocytopenia in mice","volume":"12","author":"Lien","year":"2021","journal-title":"Front Immunol"},{"key":"B20","doi-asserted-by":"publisher","first-page":"400","DOI":"10.1186\/s13104-018-3505-4","article-title":"Value of peripheral blood count for dengue severity prediction","volume":"11","author":"Ralapanawa","year":"2018","journal-title":"BMC Res Notes."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1007\/BF02725653","article-title":"Predictors of spontaneous bleeding in Dengue","volume":"71","author":"Shivbalan","year":"2004","journal-title":"Indian J Pediatr"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.10395","article-title":"Type I Glanzmann thrombasthenia: most common subtypes in North Indians","volume":"74","author":"Kannan","year":"2003","journal-title":"Am J Hematol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.4103\/ijmr.IJMR_718_14","article-title":"Evaluation of platelet surface glycoproteins in patients with Glanzmann thrombasthenia: Association with bleeding symptoms","volume":"145","author":"Mutreja","year":"2017","journal-title":"Indian J Med Res"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3390\/jcm11010118","article-title":"Platelet defects in acute myeloid leukemia-potential for hemorrhagic events","volume":"11","author":"Bumbea","year":"2021","journal-title":"J Clin Med"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1160\/TH14-01-0056","article-title":"Platelet function alterations in dengue are associated with plasma leakage","volume":"112","author":"Michels","year":"2014","journal-title":"Thromb Haemost."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2014-09-598029","article-title":"Dengue virus binding and replication by platelets","volume":"126","author":"Simon","year":"2015","journal-title":"Blood"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofx051","article-title":"Dengue virus entry and replication does not lead to productive infection in platelets","volume":"4","author":"Kar","year":"2017","journal-title":"Open Forum Infect Dis"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2019001169","article-title":"Inflammatory signaling in dengue-infected platelets requires translation and secretion of nonstructural protein 1","volume":"4","author":"Quirino-Teixeira","year":"2020","journal-title":"Blood Adv"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V94.11.3791","article-title":"Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules","volume":"94","author":"Heijnen","year":"1999","journal-title":"Blood"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1097\/MOH.0b013e32833e77ee","article-title":"Clinical relevance of microparticles from platelets and megakaryocytes","volume":"17","author":"Italiano","year":"2010","journal-title":"Curr Opin Hematol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1055\/s-0030-1267042","article-title":"Platelet- and megakaryocyte-derived microparticles","volume":"36","author":"Flaumenhaft","year":"2010","journal-title":"Semin Thromb Hemost."},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02207-14","article-title":"Microparticles provide a novel biomarker to predict severe clinical outcomes of dengue virus infection","volume":"89","author":"Punyadee","year":"2015","journal-title":"J Virol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.00481","article-title":"Contributions of mast cells and vasoactive products, leukotrienes and chymase, to dengue virus-induced vascular leakage","volume":"2","author":"St John","year":"2013","journal-title":"Elife"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.05291","article-title":"Dengue vascular leakage is augmented by mast cell degranulation mediated by immunoglobulin Fc\u03b3 receptors","volume":"4","author":"Syenina","year":"2015","journal-title":"Elife"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1172\/JCI128426","article-title":"Dengue virus-elicited tryptase induces endothelial permeability and shock","volume":"129","author":"Rathore","year":"2019","journal-title":"J Clin Invest."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23052861","article-title":"Isolation of platelet-derived exosomes from human platelet-rich plasma: biochemical and morphological characterization","volume":"23","author":"Saumell-Esnaola","year":"2022","journal-title":"Int J Mol Sci"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3083.2008.02029.x","article-title":"Soluble ICAM-1 and VCAM-1 as markers of endothelial activation","volume":"67","author":"Videm","year":"2008","journal-title":"Scand J Immunol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1093\/gerona\/glr121","article-title":"Relations of a marker of endothelial activation (s-VCAM) to function and mortality in community-dwelling older adults","volume":"66","author":"Huffman","year":"2011","journal-title":"J Gerontol A Biol Sci Med Sci"},{"key":"B39","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1002\/jmv.2007","article-title":"Plasma concentrations of sVCAM-1 and severity of dengue infections","volume":"65","author":"Murgue","year":"2001","journal-title":"J Med Virol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.20007","article-title":"Elevation of soluble VCAM-1 plasma levels in children with acute dengue virus infection of varying severity","volume":"72","author":"Koraka","year":"2004","journal-title":"J Med Virol"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1080\/10623320600972135","article-title":"Evidence of vascular damage in dengue disease: demonstration of high levels of soluble cell adhesion molecules and circulating endothelial cells","volume":"13","author":"Cardier","year":"2006","journal-title":"Endothelium"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1097\/01.inf.0000237918.85330.b9","article-title":"Elevated soluble thrombomodulin in the febrile stage related to patients at risk for dengue shock syndrome","volume":"25","author":"Butthep","year":"2006","journal-title":"Pediatr Infect Dis J"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1038\/emi.2015.24","article-title":"Serum levels of soluble vascular cell adhesion molecules may correlate with the severity of dengue virus-1 infection in adults","volume":"4","author":"Liao","year":"2015","journal-title":"Emerg Microbes Infect"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/936062","article-title":"Utility of C-reactive protein levels for early prediction of dengue severity in adults","volume":"2015","author":"Chen","year":"2015","journal-title":"BioMed Res Int"},{"key":"B45","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1186\/s12916-020-1496-1","article-title":"C-reactive protein as a potential biomarker for disease progression in dengue: a multi-country observational study","volume":"18","author":"Vuong","year":"2020","journal-title":"BMC Med"},{"key":"B46","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1038\/s41598-020-80144-0","article-title":"Increased levels of pentraxins protein and cytokines bear good association in patients with severe dengue infection","volume":"11","author":"Patra","year":"2021","journal-title":"Sci Rep"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.3VMR0315-080R","article-title":"Emerging functions of serum amyloid A in inflammation","volume":"98","author":"Ye","year":"2015","journal-title":"J Leukoc Biol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1285162\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,3]],"date-time":"2024-01-03T04:32:00Z","timestamp":1704256320000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1285162\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,3]]},"references-count":47,"alternative-id":["10.3389\/fimmu.2023.1285162"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1285162","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,3]]},"article-number":"1285162"}}