Genome editing for scalable production of alloantigen‐free lentiviral vectors for in vivo gene therapy
Open accessAbstract
Lentiviral vectors (LV) are powerful and versatile vehicles for gene therapy. However, their complex biological composition challenges large‐scale manufacturing and raises concerns for in vivo applications, because particle components and contaminants may trigger immune responses. Here, we show that producer cell‐derived polymorphic class‐I major histocompatibility complexes (MHC‐I) are incorporated into the LV surface and trigger allogeneic T‐cell responses. By disrupting the beta‐2 microglobulin gene in producer cells, we obtained MHC‐free LV with substantially reduced immunogenicity. We introduce this targeted editing into a novel stable LV packaging cell line, carrying single‐copy inducible vector components, which can be reproducibly converted into high‐yield LV producers upon site‐specific integration of the LV genome of interest. These LV efficiently transfer genes into relevant targets and are more resistant to complement‐mediated inactivation, because of reduced content of the vesicular stomatitis virus envelope glycoprotein G compared to vectors produced by transient transfection. Altogether, these advances support scalable manufacturing of alloantigen‐free LV with higher purity and increased complement resistance that are better suited for in vivo gene therapy. Lentiviral vectors (LV) lacking major histocompatibility complexes (MHC) and with low vesicular stomatitis virus glycoprotein G (VSV.G) content escape immune recognition by human T cells and complement. Cell line‐packaged LV offer scalable and consistent LV production for in vivo gene therapy. Lentiviral vectors (LV) lacking major histocompatibility complexes (MHC) and with low vesicular stomatitis virus glycoprotein G (VSV.G) content escape immune recognition by human T cells and complement. Cell line packaged LV offer scalable and consistent LV production for in vivo gene therapy.
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Gene editing of packaging cells to generate lines that lack HLA class I can enhance the stability of lentiviral vectors in serum.
“Gene editing of packaging cells to generate lines that lack HLA class I can enhance the stability of lentiviral vectors in serum [81].”
From Clinical use of lentiviral vectors (2018) · cites this paper as [81]
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Cited by
Sorted by citations
Clinical use of lentiviral vectors
Milone, M. C., & O’Doherty, U. (2018) · Leukemia · doi:10.1038/s41375-018-0106-0 · Cited by 930 · see citing works
“Gene editing of packaging cells to generate lines that lack HLA class I can enhance the stability of lentiviral vectors in serum [81].”
BackgroundImmune Responses to Viral Gene Therapy Vectors
Shirley, J. L., de Jong, Y. P., Terhorst, C., & Herzog, R. W. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.01.001 · Cited by 788 · see citing works
Biavasco, R., Lettera, E., Giannetti, K., Gilioli, D., Beretta, S., Conti, A., Scala, S., Cesana, D., Gallina, P., Norelli, M., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-24876-1 · Cited by 47 · see citing works
- Authors
- Milani, M., Annoni, A., Bartolaccini, S., Biffi, M., Russo, F., Di Tomaso, T., Raimondi, A., Lengler, J., Holmes, M. C., Scheiflinger, F., Lombardo, A., Cantore, A., & Naldini, L.
- Affiliations
- Vita-Salute San Raffaele University (IT)
- The San Raffaele Telethon Institute for Gene Therapy (IT)
- IRCCS Ospedale San Raffaele (IT)
- Baxter (Austria) (AT)
- Sangamo BioSciences (United States) (US)
- Corresponding author
- Naldini, Luigi
- ORCID
- Venue
- EMBO Molecular Medicine
- Published
- 2017-08-23
- Volume
- 9
- Issue
- 11
- Pages
- 1558-1573
- ISSN
- 1757-4676 (ISSN-L)
- 1757-4684
- PMC
- PMC5666310
- OpenAlex
- W2745793084
- Cited by
- 72
- References
- 7
- Citations by year
- 2026: 15 · 2025: 12 · 2024: 6 · 2023: 3 · 2022: 7 · 2021: 10 · 2020: 6 · 2019: 6 · 2018: 6
- FWCI
- 3.27
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- EMBO Molecular Medicine (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Archivio Istituzionale della Ricerca - Università Vita-Salute San Raffaele (submittedVersion)
- Funders
- Bioverativ
- Fondazione Telethon (TGT11D03)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- lentiviral vectors, genome editing, MHC class I, beta2-microglobulin, immunogenicity, allogeneic T cell response, complement resistance, in vivo gene therapy, site-specific integration
- MeSH terms
- Gene Editing, Animals, Cell Line, Hemophilia B, Factor IX, Genetic Vectors, Humans, Isoantigens, Mice, Inbred C57BL, Transfection, Genetic Therapy, Lentivirus, Receptors, Complement 3b, Mice, Knockout, CD55 Antigens, Mice, Membrane Cofactor Protein, HEK293 Cells
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