Nectin-2 Acts as a Viral Entry Mediated Molecule That Binds to Human Herpesvirus 6B Glycoprotein B
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Virus Strains
2.2. Reverse Transcription-PCR (RT-PCR) and Quantitative RT-PCR
2.3. Antibodies
2.4. Flow Cytometric Analysis
2.5. Immunofluorescence Assay (IFA)
2.6. Establishment of a CD134-Membrane Unanchored HSY Cell Clone (Ex6 C7)
2.7. Establishment of Nectin-2 Knockout Ex6 C7 (Ex6 C7 Nec2-KO) Cell Clones
2.8. Expression Screening
2.9. Generation of Nectin-2- and CD134-Expressing Cell Lines
2.10. Generation of Nectin-2- and CD134-Expressing Cell Lines
2.11. Transfection, Immunoblotting, and Co-IP
2.12. Nectin-2 Binding Assay in HHV-6B-Infected Cells
2.13. Assay of Nectin-2 Binding to Purified HHV-6B gB
2.14. Statistical Analysis
3. Results
3.1. HHV-6B Infects the HSY Parotid Gland Cell Line
3.2. HHV-6B Infects Ex6 C7 Cells (CD134-Membrane Unanchored HSY Cells)
3.3. Nectin-2 Is Required for HHV-6B Infection
3.4. Nectin-2 Is a Functional Entry-Mediated Molecule for HHV-6B HST and Z29 Strains
3.5. Nectin-2 Enables HHV-6B Replication in CCRF-HSB-2 Cells
3.6. Nectin-2 Interacts with HHV-6B gB
3.7. The V-set Domain of Nectin-2 Is Important for Interactions with HHV-6B gB
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Ogawa, H.; Fujikura, D.; Namba, H.; Yamashita, N.; Honda, T.; Yamada, M. Nectin-2 Acts as a Viral Entry Mediated Molecule That Binds to Human Herpesvirus 6B Glycoprotein B. Viruses 2022, 14, 160. https://doi.org/10.3390/v14010160
Ogawa H, Fujikura D, Namba H, Yamashita N, Honda T, Yamada M. Nectin-2 Acts as a Viral Entry Mediated Molecule That Binds to Human Herpesvirus 6B Glycoprotein B. Viruses. 2022; 14(1):160. https://doi.org/10.3390/v14010160
Chicago/Turabian StyleOgawa, Hirohito, Daisuke Fujikura, Hikaru Namba, Nobuko Yamashita, Tomoyuki Honda, and Masao Yamada. 2022. "Nectin-2 Acts as a Viral Entry Mediated Molecule That Binds to Human Herpesvirus 6B Glycoprotein B" Viruses 14, no. 1: 160. https://doi.org/10.3390/v14010160