Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells
Cedar virus (CedV), closely related to the Hendra and Nipah viruses, is a novel <i>Henipavirus</i> that was originally isolated from flying foxes in Australia in 2012. Although its glycoprotein G exhibits relatively low sequence similarity with its counterparts of the Hendra and Nipah vi...
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| author | Lea Lenhard Martin Müller Sandra Diederich Lisa Loerzer Virginia Friedrichs Bernd Köllner Stefan Finke Anca Dorhoi Gang Pei |
| author_facet | Lea Lenhard Martin Müller Sandra Diederich Lisa Loerzer Virginia Friedrichs Bernd Köllner Stefan Finke Anca Dorhoi Gang Pei |
| author_sort | Lea Lenhard |
| collection | DOAJ |
| description | Cedar virus (CedV), closely related to the Hendra and Nipah viruses, is a novel <i>Henipavirus</i> that was originally isolated from flying foxes in Australia in 2012. Although its glycoprotein G exhibits relatively low sequence similarity with its counterparts of the Hendra and Nipah viruses, CedV also uses ephrin receptors, i.e., ephrins B1, B2, A2 and A5, to enters human cells. Nevertheless, the entry mechanism of CedV into bat cells remains unexplored. Considering that <i>Rousettus aegyptiacus</i> (Egyptian Rousette bat, ERB) is postulated to be a reservoir host for henipaviruses, we aim to reveal the receptors utilized by CedV to enable its entry into ERB cells. To this end, we cloned the class A and B ephrins of ERB and generated CHO-K1 cells stably expressing individual ephrins. We also developed a lentivirus-based pseudovirus system containing the firefly luciferase reporter. Assessment of the luciferase activity in cells expressing single ephrins demonstrated that the ERB ephrin B1 and B2 mediated CedV pseudovirus entry. Further, we generated a recombinant CedV expressing the fluorescent protein TurboFP635 (rCedV-nTurbo635). By performing high-content microscopy and flow cytometry, we unveiled that, in addition to ephrin B1 and B2, ephrin A5 was also able to mediate rCedV-nTurbo635 entry, although to a much lesser extent. In contrast to human ephrin A2, ERB ephrin A2 failed to mediate rCedV-nTurbo635 entry. Finally, we generated ERB epithelial cells with ephrin B1 and/or ephrin B2 knockdown (KD). The entry of rCedV-nTurbo635 into ERB epithelial cells was drastically impaired by ephrin B1/B2 KD, validating the importance of ephrin B1 and B2 in its entry. Altogether, we conclude that CedV primarily employs ERB ephrin B1, B2 and, possibly, A5 for its entry into ERB cells. |
| format | Article |
| id | doaj-art-225a52f86beb4db59baf6c3249940c8b |
| institution | DOAJ |
| issn | 1999-4915 |
| language | English |
| publishDate | 2025-04-01 |
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| spelling | doaj-art-225a52f86beb4db59baf6c3249940c8b2025-08-20T03:13:48ZengMDPI AGViruses1999-49152025-04-0117457310.3390/v17040573Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat CellsLea Lenhard0Martin Müller1Sandra Diederich2Lisa Loerzer3Virginia Friedrichs4Bernd Köllner5Stefan Finke6Anca Dorhoi7Gang Pei8Institute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyInstitute of Immunology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald, GermanyCedar virus (CedV), closely related to the Hendra and Nipah viruses, is a novel <i>Henipavirus</i> that was originally isolated from flying foxes in Australia in 2012. Although its glycoprotein G exhibits relatively low sequence similarity with its counterparts of the Hendra and Nipah viruses, CedV also uses ephrin receptors, i.e., ephrins B1, B2, A2 and A5, to enters human cells. Nevertheless, the entry mechanism of CedV into bat cells remains unexplored. Considering that <i>Rousettus aegyptiacus</i> (Egyptian Rousette bat, ERB) is postulated to be a reservoir host for henipaviruses, we aim to reveal the receptors utilized by CedV to enable its entry into ERB cells. To this end, we cloned the class A and B ephrins of ERB and generated CHO-K1 cells stably expressing individual ephrins. We also developed a lentivirus-based pseudovirus system containing the firefly luciferase reporter. Assessment of the luciferase activity in cells expressing single ephrins demonstrated that the ERB ephrin B1 and B2 mediated CedV pseudovirus entry. Further, we generated a recombinant CedV expressing the fluorescent protein TurboFP635 (rCedV-nTurbo635). By performing high-content microscopy and flow cytometry, we unveiled that, in addition to ephrin B1 and B2, ephrin A5 was also able to mediate rCedV-nTurbo635 entry, although to a much lesser extent. In contrast to human ephrin A2, ERB ephrin A2 failed to mediate rCedV-nTurbo635 entry. Finally, we generated ERB epithelial cells with ephrin B1 and/or ephrin B2 knockdown (KD). The entry of rCedV-nTurbo635 into ERB epithelial cells was drastically impaired by ephrin B1/B2 KD, validating the importance of ephrin B1 and B2 in its entry. Altogether, we conclude that CedV primarily employs ERB ephrin B1, B2 and, possibly, A5 for its entry into ERB cells.https://www.mdpi.com/1999-4915/17/4/573Cedar virushenipavirusesephrinsviral entryreceptor<i>Rousettus aegyptiacus</i> |
| spellingShingle | Lea Lenhard Martin Müller Sandra Diederich Lisa Loerzer Virginia Friedrichs Bernd Köllner Stefan Finke Anca Dorhoi Gang Pei Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells Viruses Cedar virus henipaviruses ephrins viral entry receptor <i>Rousettus aegyptiacus</i> |
| title | Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells |
| title_full | Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells |
| title_fullStr | Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells |
| title_full_unstemmed | Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells |
| title_short | Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells |
| title_sort | ephrin b1 and b2 mediate cedar virus entry into egyptian fruit bat cells |
| topic | Cedar virus henipaviruses ephrins viral entry receptor <i>Rousettus aegyptiacus</i> |
| url | https://www.mdpi.com/1999-4915/17/4/573 |
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