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Article

A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions

by
Raphael L. Tsoukas
1,2,
Wolfram Volkwein
3,
Jian Gao
1,
Maren Schiwon
1,
Nora Bahlmann
1,
Thomas Dittmar
4,
Claudia Hagedorn
5,
Eric Ehrke-Schulz
1,
Wenli Zhang
1,
Armin Baiker
3 and
Anja Ehrhardt
1,*
1
Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, 58453 Witten, Germany
2
Department of Anesthesiology and Intensive Care Medicine, Medical Faculty, University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
3
Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, Germany
4
Institute of Immunology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten/Herdecke University, 58455 Witten, Germany
5
Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University (UW/H), 58448 Witten, Germany
*
Author to whom correspondence should be addressed.
Cells 2022, 11(5), 841; https://doi.org/10.3390/cells11050841
Submission received: 30 December 2021 / Revised: 11 February 2022 / Accepted: 14 February 2022 / Published: 1 March 2022
(This article belongs to the Special Issue Cell Surface Receptors)
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Abstract

To develop adenoviral cell- or tissue-specific gene delivery, understanding of the infection mechanisms of adenoviruses is crucial. Several adenoviral attachment proteins such as CD46, CAR and sialic acid have been identified and studied. However, most receptor studies were performed on non-human cells. Combining our reporter gene-tagged adenovirus library with an in vitro human gene knockout model, we performed a systematic analysis of receptor usage comparing different adenoviruses side-by-side. The CRISPR/Cas9 system was used to knockout CD46 and CAR in the human lung epithelial carcinoma cell line A549. Knockout cells were infected with 22 luciferase-expressing adenoviruses derived from adenovirus species B, C, D and E. HAdV-B16, -B21 and -B50 from species B1 as well as HAdV-B34 and -B35 were found to be CD46-dependent. HAdV-C5 and HAdV-E4 from species E were found to be CAR-dependent. Regarding cell entry of HAdV-B3 and -B14 and all species D viruses, both CAR and CD46 play a role, and here, other receptors or attachment structures may also be important since transductions were reduced but not completely inhibited. The established human knockout cell model enables the identification of the most applicable adenovirus types for gene therapy and to further understand adenovirus infection biology.
Keywords: adenovirus; receptor; CD46; CAR; knockout cell line; human; virus; tight-junction-knockout cell line; CD46-knockout cell line; CRISPR adenovirus; receptor; CD46; CAR; knockout cell line; human; virus; tight-junction-knockout cell line; CD46-knockout cell line; CRISPR

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MDPI and ACS Style

Tsoukas, R.L.; Volkwein, W.; Gao, J.; Schiwon, M.; Bahlmann, N.; Dittmar, T.; Hagedorn, C.; Ehrke-Schulz, E.; Zhang, W.; Baiker, A.; et al. A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions. Cells 2022, 11, 841. https://doi.org/10.3390/cells11050841

AMA Style

Tsoukas RL, Volkwein W, Gao J, Schiwon M, Bahlmann N, Dittmar T, Hagedorn C, Ehrke-Schulz E, Zhang W, Baiker A, et al. A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions. Cells. 2022; 11(5):841. https://doi.org/10.3390/cells11050841

Chicago/Turabian Style

Tsoukas, Raphael L., Wolfram Volkwein, Jian Gao, Maren Schiwon, Nora Bahlmann, Thomas Dittmar, Claudia Hagedorn, Eric Ehrke-Schulz, Wenli Zhang, Armin Baiker, and et al. 2022. "A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions" Cells 11, no. 5: 841. https://doi.org/10.3390/cells11050841

APA Style

Tsoukas, R. L., Volkwein, W., Gao, J., Schiwon, M., Bahlmann, N., Dittmar, T., Hagedorn, C., Ehrke-Schulz, E., Zhang, W., Baiker, A., & Ehrhardt, A. (2022). A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions. Cells, 11(5), 841. https://doi.org/10.3390/cells11050841

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