Foamy viruses (FVs) or heterologous retroviruses pseudotyped with FV glycoprotein enable transduction of a great variety of target tissues of disparate species. Specific cellular entry receptors responsible for this exceptionally broad tropism await their identification. Though, ubiquitously expressed heparan sulfate proteoglycan (HS-PG) is known to serve as an attachment factor of FV envelope (Env)-containing virus particles, greatly enhancing target cell permissiveness. Production of high-titer, FV Env-containing retroviral vectors is strongly dependent on the use of cationic polymer-based transfection reagents like polyethyleneimine (PEI). We identified packaging cell-surface HS-PG expression to be responsible for this requirement. Efficient release of FV Env-containing virus particles necessitates neutralization of HS-PG binding sites by PEI. Remarkably, remnants of PEI in FV Env-containing vector supernatants, which are not easily removable, negatively impact target cell transduction, in particular those of myeloid and lymphoid origin. To overcome this limitation for production of FV Env-containing retrovirus supernatants, we generated 293T-based packaging cell lines devoid of HS-PG by genome engineering. This enabled, for the first, time production of inhibitor-free, high-titer FV Env-containing virus supernatants by non-cationic polymer-mediated transfection. Depending on the type of virus, produced titers were 2- to 10-fold higher compared with those obtained by PEI transfection.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:91679 |
| Date | 04 June 2024 |
| Creators | Munz, Clara Marie, Kreher, Henriette, Erdbeer, Alexander, Stanke, Nicole, Richter, Stefanie, Westphal, Dana, Yi, Buqing, Behrendt, Rayk, Lindel, Fabian, Lindemann, Dirk |
| Publisher | Elsevier |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 2329-0501, 10.1016/j.omtm.2022.07.004, info:eu-repo/grantAgreement/Deutsche Forschungsgemeinschaft/SPP 1923: Innate Sensing and Restriction of Retroviruses/273898015/, info:eu-repo/grantAgreement/Deutsche Forschungsgemeinschaft/TRR 237: Nukleinsäure-Immunität/369799452//Mobile genetische Elemente als Quellen endogener immunstimulatorischer Nukleinsäuren/B19 |
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