{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T15:50:20Z","timestamp":1753890620387,"version":"3.41.2"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T00:00:00Z","timestamp":1722470400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004359","name":"Vetenskapsr\u00e5det","doi-asserted-by":"publisher","award":["2018-05795"],"award-info":[{"award-number":["2018-05795"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005753","name":"Kungliga Fysiografiska S\u00e4llskapet i Lund","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005753","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006738","name":"Medicinska Fakulteten, Lunds Universitet","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006738","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004722","name":"Stiftelsen Lars Hiertas Minne","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004722","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p><jats:italic>Streptococcus pneumoniae<\/jats:italic> (the pneumococcus) effectively colonizes the human nasopharynx, but can migrate to other host sites, causing infections such as pneumonia and sepsis. Previous studies indicate that pneumococci grown as biofilms have phenotypes of bacteria associated with colonization whereas bacteria released from biofilms in response to changes in the local environment (i.e., dispersed bacteria) represent populations with phenotypes associated with disease. How these niche-adapted populations interact with immune cells upon reaching the vascular compartment has not previously been studied. Here, we investigated neutrophil, monocyte, and platelet activation using <jats:italic>ex vivo<\/jats:italic> stimulation of whole blood and platelet-rich plasma with pneumococcal populations representing distinct stages of the infectious process (biofilm bacteria and dispersed bacteria) as well as conventional broth-grown culture (planktonic bacteria).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Flow cytometry and ELISA were used to assess surface and soluble activation markers for neutrophil and monocyte activation, platelet-neutrophil complex and platelet-monocyte complex formation, and platelet activation and responsiveness.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Overall, we found that biofilm-derived bacteria (biofilm bacteria and dispersed bacteria) induced significant activation of neutrophils, monocytes, and platelets. In contrast, little to no activation was induced by planktonic bacteria. Platelets remained functional after stimulation with bacterial populations and the degree of responsiveness was inversely related to initial activation. Bacterial association with immune cells followed a similar pattern as activation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>Differences in activation of and association with immune cells by biofilm-derived populations could be an important consideration for other pathogens that have a biofilm state. Gaining insight into how these bacterial populations interact with the host immune response may reveal immunomodulatory targets to interfere with disease development.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1405333","type":"journal-article","created":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T05:11:46Z","timestamp":1722489106000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Biofilm-dispersed pneumococci induce elevated leukocyte and platelet activation"],"prefix":"10.3389","volume":"14","author":[{"given":"Yashuan","family":"Chao","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martina","family":"M\u00f8rch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anders P.","family":"H\u00e5kansson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oonagh","family":"Shannon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,8,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1016\/j.jinf.2017.09.013","article-title":"Review manuscript: Mechanisms of platelet activation by the pneumococcus and the role of platelets in community-acquired pneumonia","volume":"75","author":"Anderson","year":"2017","journal-title":"J. Infect."},{"key":"B2","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1084\/jem.158.2.559","article-title":"Identification of an active disaccharide unit of a glycoconjugate receptor for pneumococci attaching to human pharyngeal epithelial cells","volume":"158","author":"Andersson","year":"1983","journal-title":"J. Exp. Med."},{"key":"B3","doi-asserted-by":"publisher","first-page":"3166","DOI":"10.1182\/blood-2013-11-540526","article-title":"Amplification of bacteria-induced platelet activation is triggered by FcgammaRIIA, integrin alphaIIbbeta3, and platelet factor 4","volume":"123","author":"Arman","year":"2014","journal-title":"Blood"},{"key":"B4","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1084\/jem.79.2.137","article-title":"Studies on the chemical nature of the substance inducing transformation of pneumococcal types: induction of transformation by a desoxyribonucleic acid fraction isolated from pneumococcus type iii","volume":"79","author":"Avery","year":"1944","journal-title":"J. Exp. Med."},{"key":"B5","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1016\/j.mib.2016.11.003","article-title":"Viral-bacterial co-infections in the respiratory tract","volume":"35","author":"Bakaletz","year":"2017","journal-title":"Curr. Opin. Microbiol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"4446","DOI":"10.1182\/blood.V92.11.4446","article-title":"P-Selectin and platelet clearance","volume":"92","author":"Berger","year":"1998","journal-title":"Blood"},{"key":"B7","doi-asserted-by":"publisher","first-page":"e00745","DOI":"10.1128\/mBio.00745-13","article-title":"Streptococcus pneumoniae biofilm formation is strain dependent, multifactorial, and associated with reduced invasiveness and immunoreactivity during colonization","volume":"4","author":"Blanchette-Cain","year":"2013","journal-title":"mBio"},{"key":"B8","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1016\/S1473-3099(04)00938-7","article-title":"Streptococcus pneumoniae colonization: the key to pneumococcal disease","volume":"4","author":"Bogaert","year":"2004","journal-title":"Lancet Infect. Dis."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003057","article-title":"Viral and bacterial interactions in the upper respiratory tract","volume":"9","author":"Bosch","year":"2013","journal-title":"PloS Pathog."},{"key":"B10","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1007\/978-1-4939-9199-0_13","article-title":"Growing and characterizing biofilms formed by streptococcus pneumoniae","volume":"1968","author":"Chao","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"22455","DOI":"10.1038\/s41598-020-80233-0","article-title":"The serine protease HtrA plays a key role in heat-induced dispersal of pneumococcal biofilms","volume":"10","author":"Chao","year":"2020","journal-title":"Sci. Rep."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1092","DOI":"10.1080\/09537104.2021.1921721","article-title":"Distinct phenotypes of platelet, monocyte, and neutrophil activation occur during the acute and convalescent phase of COVID-19","volume":"32","author":"Chao","year":"2021","journal-title":"Platelets"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0156977","article-title":"Toll-like receptor signaling is not involved in platelet response to streptococcus pneumoniae in vitro or in vivo","volume":"11","author":"de Stoppelaar","year":"2016","journal-title":"PloS One"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2016.00155","article-title":"The danger signal extracellular ATP is an inducer of fusobacterium nucleatum biofilm dispersal","volume":"6","author":"Ding","year":"2016","journal-title":"Front. Cell Infect. Microbiol."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.615959","article-title":"The role of neutrophils and neutrophil elastase in pneumococcal pneumonia","volume":"11","author":"Domon","year":"2021","journal-title":"Front. Cell Infect. Microbiol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1038\/nrmicro.2016.94","article-title":"Biofilms: an emergent form of bacterial life","volume":"14","author":"Flemming","year":"2016","journal-title":"Nat. Rev. Microbiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"20218","DOI":"10.1073\/pnas.1313905110","article-title":"Inducing host protection in pneumococcal sepsis by preactivation of the Ashwell-Morell receptor","volume":"110","author":"Grewal","year":"2013","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B18","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1038\/s41522-019-0098-1","article-title":"Colonization and immune modulation properties of Klebsiella pneumoniae biofilm-dispersed cells","volume":"5","author":"Guilhen","year":"2019","journal-title":"NPJ Biofilms Microbiomes"},{"key":"B19","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1001\/jama.296.2.202","article-title":"Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media","volume":"296","author":"Hall-Stoodley","year":"2006","journal-title":"JAMA"},{"key":"B20","doi-asserted-by":"publisher","first-page":"2141","DOI":"10.1111\/jth.13147","article-title":"Platelet secretory behavior: as diverse as the granules \u2026 or not","volume":"13","author":"Heijnen","year":"2015","journal-title":"J. Thromb. Haemost."},{"key":"B21","doi-asserted-by":"publisher","first-page":"1242","DOI":"10.1016\/j.ijporl.2009.05.016","article-title":"Identification of adenoid biofilms with middle ear pathogens in otitis-prone children utilizing SEM and FISH","volume":"73","author":"Hoa","year":"2009","journal-title":"Int. J. Pediatr. Otorhinolaryngol"},{"key":"B22","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/S0021-9258(18)65431-6","article-title":"The interaction of quinacrine with adenine nucleotides","volume":"210","author":"Irvin","year":"1954","journal-title":"J. Biol. Chem."},{"key":"B23","doi-asserted-by":"publisher","first-page":"1464","DOI":"10.1111\/jth.15703","article-title":"alpha-hemolysin of Staphylococcus aureus impairs thrombus formation","volume":"20","author":"Jahn","year":"2022","journal-title":"J. Thromb. Haemost."},{"key":"B24","doi-asserted-by":"publisher","first-page":"6315","DOI":"10.1182\/bloodadvances.2020002372","article-title":"Pneumolysin induces platelet destruction, not platelet activation, which can be prevented by immunoglobulin preparations in vitro","volume":"4","author":"Jahn","year":"2020","journal-title":"Blood Adv."},{"key":"B25","doi-asserted-by":"publisher","first-page":"1121","DOI":"10.3390\/cells11071121","article-title":"Platelets, bacterial adhesins and the pneumococcus","volume":"11","author":"Jahn","year":"2022","journal-title":"Cells"},{"key":"B26","doi-asserted-by":"publisher","first-page":"977","DOI":"10.1016\/j.jaci.2016.04.062","article-title":"Airborne dust and high temperatures are risk factors for invasive bacterial disease","volume":"139","author":"Jusot","year":"2017","journal-title":"J. Allergy Clin. Immunol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"5579","DOI":"10.4049\/jimmunol.1500259","article-title":"Nouvelle cuisine: platelets served with inflammation","volume":"194","author":"Kapur","year":"2015","journal-title":"J. Immunol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1846","DOI":"10.3390\/microorganisms9091846","article-title":"Planktonic and Biofilm-Associated Pseudomonas aeruginosa and Staphylococcus epidermidis Elicit Differential Human Peripheral Blood Cell Responses","volume":"9","author":"Kaya","year":"2021","journal-title":"Microorganisms"},{"key":"B29","doi-asserted-by":"publisher","first-page":"2757","DOI":"10.1111\/j.1538-7836.2010.04093.x","article-title":"Invasive Streptococcus pneumoniae trigger platelet activation via Toll-like receptor 2","volume":"8","author":"Keane","year":"2010","journal-title":"J. Thromb. Haemost."},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00213-18","article-title":"Desialylation of platelets by pneumococcal neuraminidase A induces ADP-dependent platelet hyperreactivity","volume":"86","author":"Kullaya","year":"2018","journal-title":"Infect. Immun."},{"key":"B31","doi-asserted-by":"publisher","first-page":"3581","DOI":"10.3390\/cells10123581","article-title":"Neutrophil-derived extracellular vesicles activate platelets after pneumolysin exposure","volume":"10","author":"Letsiou","year":"2021","journal-title":"Cells"},{"key":"B32","doi-asserted-by":"publisher","first-page":"2341","DOI":"10.1161\/ATVBAHA.110.207522","article-title":"Signaling during platelet adhesion and activation","volume":"30","author":"Li","year":"2010","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.GPP3-0004-2018","article-title":"Streptococcus pneumoniae: invasion and inflammation","volume":"7","author":"Loughran","year":"2019","journal-title":"Microbiol. Spectr."},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00438-13","article-title":"Interkingdom signaling induces Streptococcus pneumoniae biofilm dispersion and transition from asymptomatic colonization to disease","volume":"4","author":"Marks","year":"2013","journal-title":"mBio"},{"key":"B35","doi-asserted-by":"publisher","first-page":"2744","DOI":"10.1128\/IAI.00488-12","article-title":"Pneumococcal interactions with epithelial cells are crucial for optimal biofilm formation and colonization in vitro and in vivo","volume":"80","author":"Marks","year":"2012","journal-title":"Infect. Immun."},{"key":"B36","doi-asserted-by":"publisher","first-page":"11877","DOI":"10.1073\/pnas.93.21.11877","article-title":"In vivo tracking of platelets: circulating degranulated platelets rapidly lose surface P-selectin but continue to circulate and function","volume":"93","author":"Michelson","year":"1996","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B37","doi-asserted-by":"publisher","first-page":"5049","DOI":"10.1128\/IAI.00425-08","article-title":"Isolation of Streptococcus pneumoniae biofilm mutants and their characterization during nasopharyngeal colonization","volume":"76","author":"Munoz-Elias","year":"2008","journal-title":"Infect. Immun."},{"key":"B38","doi-asserted-by":"publisher","first-page":"599","DOI":"10.1016\/j.jinf.2017.02.010","article-title":"Pneumolysin mediates heterotypic aggregation of neutrophils and platelets in vitro","volume":"74","author":"Nel","year":"2017","journal-title":"J. Infect."},{"key":"B39","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1111\/j.1538-7836.2008.03219.x","article-title":"Elevated platelet and leukocyte response to oral bacteria in periodontitis","volume":"7","author":"Nicu","year":"2009","journal-title":"J. Thromb. Haemost."},{"key":"B40","doi-asserted-by":"publisher","first-page":"893","DOI":"10.1016\/S0140-6736(09)61204-6","article-title":"Burden of disease caused by Streptococcus pneumoniae in children younger than 5 years: global estimates","volume":"374","author":"O\u2019Brien","year":"2009","journal-title":"Lancet"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-8-206","article-title":"Proinflammatory effect in whole blood by free soluble bacterial components released from planktonic and biofilm cells","volume":"8","author":"Oscarsson","year":"2008","journal-title":"BMC Microbiol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"302","DOI":"10.1055\/s-0040-1708827","article-title":"A champion of host defense: A generic large-scale cause for platelet dysfunction and depletion in infection","volume":"46","author":"Page","year":"2020","journal-title":"Semin. Thromb. Hemost"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00462-21","article-title":"Distinct serotypes of streptococcal M proteins mediate fibrinogen-dependent platelet activation and proinflammatory effects","volume":"90","author":"Palm","year":"2022","journal-title":"Infect. Immun."},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2017.00160","article-title":"With friends like these: the complex role of neutrophils in the progression of severe pneumonia","volume":"7","author":"Pechous","year":"2017","journal-title":"Front. Cell Infect. Microbiol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"4607","DOI":"10.1128\/IAI.02225-14","article-title":"Dynamic changes in the Streptococcus pneumoniae transcriptome during transition from biofilm formation to invasive disease upon influenza A virus infection","volume":"82","author":"Pettigrew","year":"2014","journal-title":"Infect. Immun."},{"key":"B46","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1093\/cvr\/cvv048","article-title":"Platelets in leucocyte recruitment and function","volume":"107","author":"Rossaint","year":"2015","journal-title":"Cardiovasc. Res."},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2022.111614","article-title":"Platelet p110beta mediates platelet-leukocyte interaction and curtails bacterial dissemination in pneumococcal pneumonia","volume":"41","author":"Schrottmaier","year":"2022","journal-title":"Cell Rep."},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2023.2254599","article-title":"Staphylococcus aureus planktonic but not biofilm environment induces an IFN-beta macrophage immune response via the STING\/IRF3 pathway","volume":"14","author":"Seebach","year":"2023","journal-title":"Virulence"},{"key":"B49","doi-asserted-by":"publisher","first-page":"236","DOI":"10.12688\/f1000research.13283.1","article-title":"The life cycle of platelet granules","volume":"7","author":"Sharda","year":"2018","journal-title":"F1000Res"},{"key":"B50","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/B978-0-12-410530-0.00015-6","article-title":"Nasopharyngeal colonization with Streptococcus pneumoniae","volume-title":"Streptococcus pneumoniae: molecular mechanisms of host-pathogen interactions","author":"Short","year":"2015"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00255-12","article-title":"Increased nasopharyngeal bacterial titers and local inflammation facilitate transmission of Streptococcus pneumoniae","volume":"3","author":"Short","year":"2012","journal-title":"mBio"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006990","article-title":"Shiga toxin and lipopolysaccharide induce platelet-leukocyte aggregates and tissue factor release, a thrombotic mechanism in hemolytic uremic syndrome","volume":"4","author":"Stahl","year":"2009","journal-title":"PloS One"},{"key":"B53","doi-asserted-by":"publisher","first-page":"704","DOI":"10.1016\/j.chom.2015.03.008","article-title":"The infant nasopharyngeal microbiome impacts severity of lower respiratory infection and risk of asthma development","volume":"17","author":"Teo","year":"2015","journal-title":"Cell Host Microbe"},{"key":"B54","doi-asserted-by":"publisher","first-page":"7465","DOI":"10.1073\/pnas.1815572116","article-title":"Dual actions of group B Streptococcus capsular sialic acid provide resistance to platelet-mediated antimicrobial killing","volume":"116","author":"Uchiyama","year":"2019","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B55","doi-asserted-by":"publisher","first-page":"e75","DOI":"10.1097\/CCM.0000000000000853","article-title":"Thrombocytopenia impairs host defense during murine Streptococcus pneumoniae pneumonia","volume":"43","author":"van den Boogaard","year":"2015","journal-title":"Crit. Care Med."},{"key":"B56","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1111\/j.1365-2141.1995.tb03315.x","article-title":"A flow cytometric assay using mepacrine for study of uptake and release of platelet dense granule contents","volume":"89","author":"Wall","year":"1995","journal-title":"Br. J. Haematol"},{"key":"B57","doi-asserted-by":"publisher","first-page":"1147","DOI":"10.1055\/a-1723-1880","article-title":"Polyvalent immunoglobulin preparations inhibit pneumolysin-induced platelet destruction","volume":"122","author":"Wiebe","year":"2022","journal-title":"Thromb. Haemost."},{"key":"B58","doi-asserted-by":"publisher","first-page":"1459","DOI":"10.1111\/jth.14814","article-title":"Activated platelets kill Staphylococcus aureus, but not Streptococcus pneumoniae-The role of FcgammaRIIa and platelet factor 4\/heparinantibodies","volume":"18","author":"Wolff","year":"2020","journal-title":"J. Thromb. Haemost."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1405333\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T05:11:53Z","timestamp":1722489113000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1405333\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,1]]},"references-count":58,"alternative-id":["10.3389\/fcimb.2024.1405333"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2024.1405333","relation":{},"ISSN":["2235-2988"],"issn-type":[{"type":"electronic","value":"2235-2988"}],"subject":[],"published":{"date-parts":[[2024,8,1]]},"article-number":"1405333"}}