Modelling enhanced phagocytosis of activated macrophage by outer membrane vesicles from Neisseria gonorrhoeae

Modelling enhanced phagocytosis of activated macrophage by outer membrane vesicles from Neisseria gonorrhoeae | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Modelling enhanced phagocytosis of activated macrophage by outer membrane vesicles from Neisseria gonorrhoeae Jiaru Yang, Shilian Xu, Thomas Naderer This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7577586/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Gonorrhoea, caused by Neisseria gonorrhoeae, remains a significant health concern. N. gonorrhoeae produces outer membrane vesicles (NOMVs) that modulate the host immune system by delivering virulence factors. Macrophages are crucial for eliminating foreign material through phagocytosis; dysfunction in this process can lead to infections. Mathematical models offer insights into bacteria-phagocyte interactions, where infection size can exhibit bistable effects. It's unclear if naïve or NOMV-activated macrophages induce bistable bacterial kinetics, which is critical for understanding if activated macrophages can alter infection outcomes, potentially aiding interventions without antimicrobials. Methods: We activated macrophages with NOMVs and used live cell imaging (LCI) assay to monitor activated cell survival during N. gonorrhoeae and E. coli infection. We developed a family of mathematical models to quantify bacterial kinetics, considering macrophage concentration, inoculum size, natural macrophage death, phagocytosis efficiency, and comparing antibacterial capabilities of trained and naïve macrophages. Results: LCI data showed NOMV activation doesn't increase macrophage natural death. Phagocytosis efficiency increases and saturates with higher macrophage and bacteria concentrations. Activation notably enhances phagocytosis efficiency, demonstrating strain-specific effects and reducing phagocytosis-associated macrophage death. Both activated and naïve macrophages induced bistable bacterial kinetics for E. coli and N. gonorrhoeae: high inoculum sizes persisted, while low inoculum sizes were inhibited. Significantly, the antibacterial capability of activated macrophages was 14.2 times stronger for E. coli and 12.4 times for N. gonorrhoeae than that of naïve macrophages, aligning with observed 10- to 20-fold increases. Conclusions: Our findings highlight the critical role of NOMV activation in enhancing macrophage antibacterial capability, offering a potential mechanistic explanation for developing N. gonorrhoeae OMV vaccines. Health sciences/Diseases Biological sciences/Immunology Biological sciences/Microbiology Neisseria gonorrhoeae macrophage outer membrane vesicles mathematical model trained immunity live cell imaging Full Text Additional Declarations No competing interests reported. Supplementary Files SupplimentaryMaterialV2SX.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7577586","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":529088732,"identity":"c9c0c1a2-0a04-4e18-a09c-9052d289c34d","order_by":0,"name":"Jiaru Yang","email":"","orcid":"","institution":"Monash University","correspondingAuthor":false,"prefix":"","firstName":"Jiaru","middleName":"","lastName":"Yang","suffix":""},{"id":529088734,"identity":"8dae3c60-da6e-40e5-8f3b-3cd64c961a56","order_by":1,"name":"Shilian Xu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/ElEQVRIiWNgGAWjYDACCeYGEJUA5doAMWPjAfxaGFG0pIG0NJCk5TCYxKtFfnZj46MbFQx5/NLHL7/4uOe83dr2w0BbamyicWkxuHOw2TjnDEOxZF9OmeWMZ7eTt51JBGo5lpbbgEuLRGKbdG4bQ+KGMzxpxjwHbiebHQBqYWw4jFOL/Ayolv0gLX8OnEs2O/8QvxaGGzBbeNgPP2Y4cMDO7AYBWwxuJIL8IlEscYaHjbHnQHKC2Q2gLQl4/CI/I/ng45wKmzz+HvbHH34csLM3O5/+8MGHGhvcDoMACSDmMQORiWCVCfiVwwDQFiBpT5ziUTAKRsEoGEkAAFPkZ+UAO2tFAAAAAElFTkSuQmCC","orcid":"","institution":"Technion—Israel Institute of Technology","correspondingAuthor":true,"prefix":"","firstName":"Shilian","middleName":"","lastName":"Xu","suffix":""},{"id":529088735,"identity":"aaa67a1b-f421-4168-95a2-21e786777b8d","order_by":2,"name":"Thomas Naderer","email":"","orcid":"","institution":"Monash University","correspondingAuthor":false,"prefix":"","firstName":"Thomas","middleName":"","lastName":"Naderer","suffix":""}],"badges":[],"createdAt":"2025-09-10 01:08:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7577586/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7577586/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":93530618,"identity":"a857d530-0fbc-4549-bf73-62ad70ad6193","added_by":"auto","created_at":"2025-10-14 20:59:08","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":3914183,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptText.docx","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/1b5c0aba0318010b79291e00.docx"},{"id":93529589,"identity":"7bdc7bd2-5e72-445f-863f-c913901d7392","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5672,"visible":true,"origin":"","legend":"","description":"","filename":"f56b58ba1cc248f5b108076e2dc75158.json","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/836edb0baca716a508189fa0.json"},{"id":93529603,"identity":"f1c250a4-7116-4b01-9e7e-984e86b8db21","added_by":"auto","created_at":"2025-10-14 20:51:09","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":4116385,"visible":true,"origin":"","legend":"","description":"","filename":"SupplimentaryMaterialV2SX.docx","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/059edd9d2610788647990063.docx"},{"id":93529592,"identity":"3dbac15b-c0ae-4507-a535-d1288fd8b2e8","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"xml","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":135732,"visible":true,"origin":"","legend":"","description":"","filename":"f56b58ba1cc248f5b108076e2dc751581enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/a7fa4690824596c96fd79faf.xml"},{"id":93529595,"identity":"01ac0921-c7a0-4190-8e01-663450d7e0b0","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"jpeg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":887660,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/66bfa705dc0c79d5e340fc14.jpeg"},{"id":93529590,"identity":"750684aa-9088-4250-919c-c11606843f98","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"jpeg","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":795044,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/b87ffdd4fc155359f4dd8623.jpeg"},{"id":93530620,"identity":"084205e3-d0d7-4d5b-855d-c6ed8733bb27","added_by":"auto","created_at":"2025-10-14 20:59:08","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":376793,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/32a1095f8e964c51fc4501a7.jpeg"},{"id":93530617,"identity":"612aec55-d21c-44a3-a819-81aaea4ade46","added_by":"auto","created_at":"2025-10-14 20:59:08","extension":"jpeg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":885680,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/31babf3cd0db83303ac3609d.jpeg"},{"id":93529594,"identity":"ba262cde-4e83-4dc1-86af-3ea4e7c67acc","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"jpeg","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":884034,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/2e22a4fafab021a072a1ebeb.jpeg"},{"id":93530616,"identity":"2b10ceab-4c03-49eb-84df-11872dd856f1","added_by":"auto","created_at":"2025-10-14 20:59:08","extension":"png","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":60866,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/00a4745cd3168b66b9ded74b.png"},{"id":93529597,"identity":"bab9a263-99d8-48a7-a713-1a2ecf3a0512","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":55395,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/58731ddcd8e0ecd050309426.png"},{"id":93529599,"identity":"6d91717b-f811-47e7-898f-3b37519da0f4","added_by":"auto","created_at":"2025-10-14 20:51:08","extension":"png","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":111485,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/67d3eaadf0b53806f0511905.png"},{"id":93529601,"identity":"9bdb3e2f-23fd-46a5-a40b-e47c5921ca08","added_by":"auto","created_at":"2025-10-14 20:51:09","extension":"png","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":71283,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/3687d28e3befda876c216f1d.png"},{"id":93530832,"identity":"afbeb72b-ede5-4e78-8393-9920f6d385ee","added_by":"auto","created_at":"2025-10-14 21:07:08","extension":"png","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":69098,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/ee5c657789e64f35b5bd188c.png"},{"id":93529605,"identity":"3e88aebf-d574-405e-b3a3-b31c8d77a556","added_by":"auto","created_at":"2025-10-14 20:51:09","extension":"xml","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":134369,"visible":true,"origin":"","legend":"","description":"","filename":"f56b58ba1cc248f5b108076e2dc751581structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/74ea3e7c21d2f34b90c609ca.xml"},{"id":93529604,"identity":"aeb90803-2d00-4bde-a393-2a188643a3dc","added_by":"auto","created_at":"2025-10-14 20:51:09","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":153074,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/933c3c40b8faa3a6841516a8.html"},{"id":97672700,"identity":"5b7a1479-1fa4-4282-b12e-6a74533acef2","added_by":"auto","created_at":"2025-12-08 09:38:35","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":982712,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptText.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1_covered_b5d1496d-4961-47eb-a76f-3d5355ea97c8.pdf"},{"id":93529602,"identity":"4c37fbdb-3dcb-443f-b433-0be8139f0e7f","added_by":"auto","created_at":"2025-10-14 20:51:09","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":4116385,"visible":true,"origin":"","legend":"","description":"","filename":"SupplimentaryMaterialV2SX.docx","url":"https://assets-eu.researchsquare.com/files/rs-7577586/v1/54ad5e6a81b0576e457b0a53.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Modelling enhanced phagocytosis of activated macrophage by outer membrane vesicles from Neisseria gonorrhoeae","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Neisseria gonorrhoeae, macrophage, outer membrane vesicles, mathematical model, trained immunity, live cell imaging","lastPublishedDoi":"10.21203/rs.3.rs-7577586/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7577586/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003eGonorrhoea, caused by Neisseria gonorrhoeae, remains a significant health concern. N. gonorrhoeae produces outer membrane vesicles (NOMVs) that modulate the host immune system by delivering virulence factors. Macrophages are crucial for eliminating foreign material through phagocytosis; dysfunction in this process can lead to infections. Mathematical models offer insights into bacteria-phagocyte interactions, where infection size can exhibit bistable effects. It's unclear if naïve or NOMV-activated macrophages induce bistable bacterial kinetics, which is critical for understanding if activated macrophages can alter infection outcomes, potentially aiding interventions without antimicrobials.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003eWe activated macrophages with NOMVs and used live cell imaging (LCI) assay to monitor activated cell survival during N. gonorrhoeae and E. coli infection. We developed a family of mathematical models to quantify bacterial kinetics, considering macrophage concentration, inoculum size, natural macrophage death, phagocytosis efficiency, and comparing antibacterial capabilities of trained and naïve macrophages.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003eLCI data showed NOMV activation doesn't increase macrophage natural death. Phagocytosis efficiency increases and saturates with higher macrophage and bacteria concentrations. Activation notably enhances phagocytosis efficiency, demonstrating strain-specific effects and reducing phagocytosis-associated macrophage death. Both activated and naïve macrophages induced bistable bacterial kinetics for E. coli and N. gonorrhoeae: high inoculum sizes persisted, while low inoculum sizes were inhibited. Significantly, the antibacterial capability of activated macrophages was 14.2 times stronger for E. coli and 12.4 times for N. gonorrhoeae than that of naïve macrophages, aligning with observed 10- to 20-fold increases.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eOur findings highlight the critical role of NOMV activation in enhancing macrophage antibacterial capability, offering a potential mechanistic explanation for developing N. gonorrhoeae OMV vaccines.\u003c/p\u003e","manuscriptTitle":"Modelling enhanced phagocytosis of activated macrophage by outer membrane vesicles from Neisseria gonorrhoeae","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-14 20:51:03","doi":"10.21203/rs.3.rs-7577586/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"40c28199-b684-4619-9e18-603ceedf11cb","owner":[],"postedDate":"October 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":56235467,"name":"Health sciences/Diseases"},{"id":56235468,"name":"Biological sciences/Immunology"},{"id":56235469,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2025-12-06T06:54:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-14 20:51:03","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7577586","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7577586","identity":"rs-7577586","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}