Improved targeting and elicitation of VRC01-class precursors by an optimized glycan-masking mutant of OD-GT8

Improved targeting and elicitation of VRC01-class precursors by an optimized glycan-masking mutant of OD-GT8 | 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 Improved targeting and elicitation of VRC01-class precursors by an optimized glycan-masking mutant of OD-GT8 Ted Pierson, Xuejun Chen, Jeffrey Boyington, Madhu Prabhakaran, and 12 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6941578/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract VRC01-class broadly neutralizing antibodies (bnAbs) target the conserved CD4-binding site (CD4bs) on the HIV-1 envelope. An engineered envelope outer domain eOD-GT8 60mer nanoparticle elicits VRC01-class precursors but also significant off-target responses in transgenic mice and humans, limiting on-target bnAb development. To mask off-target epitopes, N-linked glycans were engineered into eOD-GT8 to further focus immunogenicity to the CD4bs. Here, we developed an improved, highly glycosylated eOD-GT8 variant (eOD-GT8.Mut49) that retains affinity to all tested VRC01-class precursors but does not bind off-target antibodies. Mut49 more efficiently labels VRC01-class B cell precursors in naïve human PBMCs than eOD-GT8. Moreover, Mut49 60mer elicits higher levels of CD4bs-specific responses and VRC01-class precursors than eOD-GT8 60mer and its other derivatives in multiple transgenic mouse models expressing physiologically low frequencies of VRC01-class precursors. These studies support the further development of Mut49 as a priming immunogen in germline-targeting HIV-1 vaccine regimens. Biological sciences/Immunology/Vaccines/Protein vaccines Health sciences/Diseases/Infectious diseases/HIV infections Full Text Additional Declarations Yes there is potential Competing Interest. NIH holds a patent application titled "GLYCAN-MASKED ENGINEERED OUTER DOMAINS OF HIV-1 gp120 AND THEIR USE" that is under review by US and European Patent Offices. Patent No.: US 12,053,520B2 and EU E-083-2017-0-EP-04; Applicant: NIH; Invertors: Xuejun Chen, Jeffrey Boyington, Hongying Duan, Cheng Cheng and John Mascola. The eOD-GT8 glycan-masking mutants including mut49 described in the current manuscript is covered by this patent. Cite Share Download PDF Status: Under Review 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. 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Patent No.: US 12,053,520B2 and EU E-083-2017-0-EP-04; Applicant: NIH; Invertors: Xuejun Chen, Jeffrey Boyington, Hongying Duan, Cheng Cheng and John Mascola. The eOD-GT8 glycan-masking mutants including mut49 described in the current manuscript is covered by this patent.","formattedTitle":"Improved targeting and elicitation of VRC01-class precursors by an optimized glycan-masking mutant of OD-GT8","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"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":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6941578/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6941578/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"VRC01-class broadly neutralizing antibodies (bnAbs) target the conserved CD4-binding site (CD4bs) on the HIV-1 envelope. An engineered envelope outer domain eOD-GT8 60mer nanoparticle elicits VRC01-class precursors but also significant off-target responses in transgenic mice and humans, limiting on-target bnAb development. To mask off-target epitopes, N-linked glycans were engineered into eOD-GT8 to further focus immunogenicity to the CD4bs. Here, we developed an improved, highly glycosylated eOD-GT8 variant (eOD-GT8.Mut49) that retains affinity to all tested VRC01-class precursors but does not bind off-target antibodies. Mut49 more efficiently labels VRC01-class B cell precursors in naïve human PBMCs than eOD-GT8. Moreover, Mut49 60mer elicits higher levels of CD4bs-specific responses and VRC01-class precursors than eOD-GT8 60mer and its other derivatives in multiple transgenic mouse models expressing physiologically low frequencies of VRC01-class precursors. 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