Extraction, structural characterization, and potential immunologic adjuvant activity of polysaccharides from Polygonum cuspidatum Sieb.et Zucc against influenza virus

Extraction, structural characterization, and potential immunologic adjuvant activity of polysaccharides from Polygonum cuspidatum Sieb.et Zucc against influenza virus | 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 Research Article Extraction, structural characterization, and potential immunologic adjuvant activity of polysaccharides from Polygonum cuspidatum Sieb.et Zucc against influenza virus Xinxin Liu, Tongchao Ji, Weixia Guo, Junfang Liu, Lihua Han This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9197211/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：Polysaccharides represent essential bioactive compounds derived from Polygonum cuspidatum Sieb.et Zucc . The present investigation aimed to enhance polysaccharide extraction efficiency, achieve purification, elucidate structural characteristics, and determine their potential application as immune adjuvants against influenza virus. Results：An acid-based extraction method was employed for isolating polysaccharides from P. cuspidatum, with optimized extraction parameters established at a liquid-to-solid ratio of 20 mL/g, extraction temperature of 84 °C, duration of 50 minutes, and HCl concentration at 0.1 M. The purified fraction obtained was designated as PCPs. Structural elucidation of PCPs involved comprehensive analytical approaches. Results indicated that PCPs comprised glucans featuring a backbone structure of →4)-α-D-Glcp-(1→ residues, branched by →4,6)-α-D-Glcp-(1→ to →6)-α-D-Glcp-(1→ side chains. The O-6 position was confirmed as the branching site, and reducing terminal residues included both →4)-α-D-Glcp and →4)-β-D-Glcp. To evaluate the immunological adjuvant properties of PCPs for H1N1 influenza vaccines, mice received immunizations containing PCPs mixed with inactivated H1N1 antigen. To comprehensively evaluate the immune-enhancing properties of PCPs, we assessed specific IgG antibodies and subclasses (IgG1 and IgG2a) in serum, activation of splenic lymphocyte-derived CD4+ and CD8+ T cells, differentiation of central memory T-cell subsets characterized by the CD44 high CD62 Lhigh phenotype, and Th1/Th2 cytokine secretion profiles. Compared with mice receiving no adjuvant or antigen only, the PCP-immunized group demonstrated markedly increased antibody production of IgG2a (indicative of a Th1-type response) and IgG1 (indicative of a Th2-type response), significantly enhanced proliferation of CD4+ T cells, elevated numbers of CD8+ central memory T cells, and heightened secretion of the cytokines IFN-γ, IL-2, IL-4, IL-6, and IL-10. Conclusion：Collectively, these observations reveal that PCPs effectively boost both cellular and humoral immune responses as vaccine adjuvants against H1N1, emphasizing their promising applicability in veterinary vaccine formulations for livestock and poultry. Polygonum cuspidatum Sieb.et Zucc polysaccharide vaccine adjuvant Full Text Additional Declarations No competing interests reported. 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. 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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-9197211","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":614036094,"identity":"6a640fb3-d9a5-4e4f-b4d3-b69c1a42dd97","order_by":0,"name":"Xinxin Liu","email":"","orcid":"","institution":"Nanyang Zhang Zhongjing Hospital，Zhengzhou","correspondingAuthor":false,"prefix":"","firstName":"Xinxin","middleName":"","lastName":"Liu","suffix":""},{"id":614036095,"identity":"f80c7cb6-8137-4575-b9b7-b31572dd0c7c","order_by":1,"name":"Tongchao Ji","email":"","orcid":"","institution":"Nanyang Zhang Zhongjing Hospital，Zhengzhou","correspondingAuthor":false,"prefix":"","firstName":"Tongchao","middleName":"","lastName":"Ji","suffix":""},{"id":614036096,"identity":"c1223980-8761-4ec8-a5e5-3c46b32b57c2","order_by":2,"name":"Weixia Guo","email":"","orcid":"","institution":"Nanyang Zhang Zhongjing Hospital，Zhengzhou","correspondingAuthor":false,"prefix":"","firstName":"Weixia","middleName":"","lastName":"Guo","suffix":""},{"id":614036097,"identity":"1a2d6c85-2423-4c34-a31c-8299e8d1fdba","order_by":3,"name":"Junfang Liu","email":"","orcid":"","institution":"Nanyang Zhang Zhongjing Hospital，Zhengzhou","correspondingAuthor":false,"prefix":"","firstName":"Junfang","middleName":"","lastName":"Liu","suffix":""},{"id":614036098,"identity":"b97d042b-faa3-45f1-8809-984cc4d075c9","order_by":4,"name":"Lihua Han","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIiWNgGAWjYBACA3YeEFkjJ8/e//xD4j82O35m5sMP8GphBmkpOGZs2HOGjeEDG1+yZDtbmgFhLR+YExtu5LAxzmCTY9xwnkdBAp8Wc2begx9+GLAlNvacPfaYh8eM2fgwD4MBQ41NNC4tls1Ap/QYyBi3s/elG/NIpPGZHeY98IDhWFpuAy6HHeYxkOAxYJNt7DlgIM1jcIzZ7DBfggFjw2F8Wox//jFgZmy4kQDUkvCfcXMz0BACWsyAhjMrAr1vJjnjABvjBmYitFjLGIAC+ViywccGtmSJw8BATsDnl+M9xjff/AFFZfPBB4kNwKjsP3z4wYcaG5xacIAE0pSPglEwCkbBKEADAK2KWiPVCOdsAAAAAElFTkSuQmCC","orcid":"","institution":"Nanyang Zhang Zhongjing Hospital，Zhengzhou","correspondingAuthor":true,"prefix":"","firstName":"Lihua","middleName":"","lastName":"Han","suffix":""}],"badges":[],"createdAt":"2026-03-23 07:55:04","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9197211/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9197211/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106284334,"identity":"abb02a80-dbdc-4a4d-b98e-4fd24b7c8115","added_by":"auto","created_at":"2026-04-07 06:42:45","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1025070,"visible":true,"origin":"","legend":"","description":"","filename":"HIN122.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9197211/v1_covered_a81afd43-6136-41df-88a0-8a6fd6a4f3cf.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Extraction, structural characterization, and potential immunologic adjuvant activity of polysaccharides from Polygonum cuspidatum Sieb.et Zucc against influenza virus","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":"Polygonum cuspidatum Sieb.et Zucc, polysaccharide, vaccine adjuvant","lastPublishedDoi":"10.21203/rs.3.rs-9197211/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9197211/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground：Polysaccharides represent essential bioactive compounds derived from \u003cem\u003ePolygonum cuspidatum Sieb.et Zucc\u003c/em\u003e. The present investigation aimed to enhance polysaccharide extraction efficiency, achieve purification, elucidate structural characteristics, and determine their potential application as immune adjuvants against influenza virus.\u003c/p\u003e\n\u003cp\u003eResults：An acid-based extraction method was employed for isolating polysaccharides from P. cuspidatum, with optimized extraction parameters established at a liquid-to-solid ratio of 20 mL/g, extraction temperature of 84 °C, duration of 50 minutes, and HCl concentration at 0.1 M. The purified fraction obtained was designated as PCPs. Structural elucidation of PCPs involved comprehensive analytical approaches. Results indicated that PCPs comprised glucans featuring a backbone structure of →4)-α-D-Glcp-(1→ residues, branched by →4,6)-α-D-Glcp-(1→ to →6)-α-D-Glcp-(1→ side chains. The O-6 position was confirmed as the branching site, and reducing terminal residues included both →4)-α-D-Glcp and →4)-β-D-Glcp. To evaluate the immunological adjuvant properties of PCPs for H1N1 influenza vaccines, mice received immunizations containing PCPs mixed with inactivated H1N1 antigen. To comprehensively evaluate the immune-enhancing properties of PCPs, we assessed specific IgG antibodies and subclasses (IgG1 and IgG2a) in serum, activation of splenic lymphocyte-derived CD4+ and CD8+ T cells, differentiation of central memory T-cell subsets characterized by the CD44\u003csup\u003ehigh\u003c/sup\u003eCD62\u003csup\u003eLhigh\u003c/sup\u003e phenotype, and Th1/Th2 cytokine secretion profiles. Compared with mice receiving no adjuvant or antigen only, the PCP-immunized group demonstrated markedly increased antibody production of IgG2a (indicative of a Th1-type response) and IgG1 (indicative of a Th2-type response), significantly enhanced proliferation of CD4+ T cells, elevated numbers of CD8+ central memory T cells, and heightened secretion of the cytokines IFN-γ, IL-2, IL-4, IL-6, and IL-10.\u003c/p\u003e\n\u003cp\u003eConclusion：Collectively, these observations reveal that PCPs effectively boost both cellular and humoral immune responses as vaccine adjuvants against H1N1, emphasizing their promising applicability in veterinary vaccine formulations for livestock and poultry.\u003c/p\u003e","manuscriptTitle":"Extraction, structural characterization, and potential immunologic adjuvant activity of polysaccharides from Polygonum cuspidatum Sieb.et Zucc against influenza virus","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-30 11:18:38","doi":"10.21203/rs.3.rs-9197211/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":"64a2d7e7-1f7d-46f7-958c-316e41df02d4","owner":[],"postedDate":"March 30th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-07T06:40:08+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-30 11:18:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9197211","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9197211","identity":"rs-9197211","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}