Combined effects of restriction factors and transduction adjuvants on lentiviral vector gene transfer efficacy

Combined effects of restriction factors and transduction adjuvants on lentiviral vector gene transfer efficacy | 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 Combined effects of restriction factors and transduction adjuvants on lentiviral vector gene transfer efficacy Marie DEWANNIEUX, Fatoumata FOFANA, Kathleen HAMON, Mirella MORMIN, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6702056/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 04 Nov, 2025 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract Restriction factors include various cellular proteins that detect and impede viral infections. Among them, interferon-induced transmembrane (IFITM) and serine incorporator (SERINC) proteins interfere with the infectious cycle of HIV-1. Consequently, such restriction factors can also interfere with gene transfer efficacy when using recombinant lentiviral vectors derived from HIV-1, but these parameters remain incompletely understood. Here, using overexpressing human cell lines, we investigated the effects of IFITM and SERINC proteins on key parameters in the infectivity of lentiviral vectors, e.g. the nature of the vector envelope glycoprotein pseudotype and the use of transduction adjuvants. Vectors pseudotyped with glycoproteins from vesiculoviruses were mostly insensitive to the effects of restriction factors whereas those pseudotyped with glycoproteins of retroviral origin displayed contrasted responses. IFITM2 and IFITM3 very effectively restricted Syncytin1-pseudotyped vectors. Some transduction adjuvants counteracted these effects. Cyclosporin H and also unexpectedly, Vectofusin, both reduced IFITM2 and IFITM3 protein levels. These results may rationalize the choice of pseudotypes as well as transduction conditions to use for gene transfer. Together, these parameters can strongly enhance the transduction efficacy, especially in target cells that naturally express IFITM proteins, including human hematopoietic cells that are of particular clinical interest. Biological sciences/Biotechnology/Gene delivery Biological sciences/Biotechnology/Gene therapy Biological sciences/Immunology/Innate immunity Biological sciences/Biological techniques/Gene delivery/Genetic vectors Biological sciences/Microbiology/Virology/Restriction factors Lentiviral vector gene therapy restriction factor viral transduction Full Text Additional Declarations Competing interest reported. The author A. Galy is inventor of a patent entitled " Stable pseudotyped lentiviral particles and uses thereof " published as US11993781B2. Supplementary Files 20250123SuptableqPCRprimers.pdf 1Bbottomactin.tif 1Bbottomifitm23.tif 1Btopactin.tif 1Btopifitm1.tif Cite Share Download PDF Status: Published Journal Publication published 04 Nov, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 10 Jul, 2025 Reviews received at journal 09 Jul, 2025 Reviews received at journal 01 Jul, 2025 Reviewers agreed at journal 27 Jun, 2025 Reviewers agreed at journal 26 Jun, 2025 Reviewers invited by journal 25 Jun, 2025 Editor assigned by journal 25 Jun, 2025 Editor invited by journal 25 Jun, 2025 Submission checks completed at journal 24 Jun, 2025 First submitted to journal 19 May, 2025 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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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-6702056","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":477414015,"identity":"09d21923-8451-41cf-ae1a-b9fd4a6b0710","order_by":0,"name":"Marie 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Among them, interferon-induced transmembrane (IFITM) and serine incorporator (SERINC) proteins interfere with the infectious cycle of HIV-1. Consequently, such restriction factors can also interfere with gene transfer efficacy when using recombinant lentiviral vectors derived from HIV-1, but these parameters remain incompletely understood. Here, using overexpressing human cell lines, we investigated the effects of IFITM and SERINC proteins on key parameters in the infectivity of lentiviral vectors, e.g. the nature of the vector envelope glycoprotein pseudotype and the use of transduction adjuvants. Vectors pseudotyped with glycoproteins from vesiculoviruses were mostly insensitive to the effects of restriction factors whereas those pseudotyped with glycoproteins of retroviral origin displayed contrasted responses. IFITM2 and IFITM3 very effectively restricted Syncytin1-pseudotyped vectors. Some transduction adjuvants counteracted these effects. 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