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Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine

Avian Tembusu virus (TMUV) is a novel flavivirus causing severe egg drop and fatal encephalitis in avian in Asia. In the present study, we screened the structural and functional requirements of TMUV capsid protein (CP) for viral morphogenesis using reverse genetics methods in combination with replic...

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Autores principales: He, Yu, Wang, Xiaoli, Guo, Jiaqi, Mao, Li, Zhang, Senzhao, Hu, Tao, Wang, Mingshu, Jia, Renyong, Zhu, Dekang, Liu, Mafeng, Zhao, Xinxin, Yang, Qiao, Wu, Ying, Zhang, Shaqiu, Huang, Juan, Mao, Sai, Ou, Xumin, Gao, Qun, Sun, Di, Liu, Yunya, Zhang, Ling, Yu, Yanling, Cheng, Anchun, Chen, Shun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371329/
https://www.ncbi.nlm.nih.gov/pubmed/34421904
http://dx.doi.org/10.3389/fimmu.2021.694959
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author He, Yu
Wang, Xiaoli
Guo, Jiaqi
Mao, Li
Zhang, Senzhao
Hu, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, Shaqiu
Huang, Juan
Mao, Sai
Ou, Xumin
Gao, Qun
Sun, Di
Liu, Yunya
Zhang, Ling
Yu, Yanling
Cheng, Anchun
Chen, Shun
author_facet He, Yu
Wang, Xiaoli
Guo, Jiaqi
Mao, Li
Zhang, Senzhao
Hu, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, Shaqiu
Huang, Juan
Mao, Sai
Ou, Xumin
Gao, Qun
Sun, Di
Liu, Yunya
Zhang, Ling
Yu, Yanling
Cheng, Anchun
Chen, Shun
author_sort He, Yu
collection PubMed
description Avian Tembusu virus (TMUV) is a novel flavivirus causing severe egg drop and fatal encephalitis in avian in Asia. In the present study, we screened the structural and functional requirements of TMUV capsid protein (CP) for viral morphogenesis using reverse genetics methods in combination with replicon packaging assays. TMUV-CP showed dramatic functional and structural flexibility, and even though 44 residues were removed from the N-terminus, it was still capable of packaging replicon RNA; in addition, 33 residues were deleted from the C-terminus (containing nearly the entire α4-helix), and infectious particles were still produced, although α4-α4’ is supposedly vital for CP dimerization and nucleocapsid formation. We further analyzed two mutants (ΔC20-43 and ΔC64-96 viruses) with relatively large deletions that still replicated well in BHK-21 cells. Our data indicate that internal deletions within CP impaired viral replication or assembly, resulting in attenuated virus proliferation in cells and attenuated virulence in duck embryos, and these deletion mutations are quite stable in cell culture. An in vivo assay indicated that both ΔC20-43 virus and ΔC64-96 virus were highly attenuated in ducklings but still immunogenic. Single-dose immunization with ΔC20-43 virus or ΔC64-96 virus could protect ducklings from a lethal challenge with good antigen clearance. Together, our data shed light on replication/assembly defective TMUV with internal deletions in CP and provide an effective approach to attenuate viral virulence in live vaccines without changing the antigen composition.
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spelling pubmed-83713292021-08-19 Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine He, Yu Wang, Xiaoli Guo, Jiaqi Mao, Li Zhang, Senzhao Hu, Tao Wang, Mingshu Jia, Renyong Zhu, Dekang Liu, Mafeng Zhao, Xinxin Yang, Qiao Wu, Ying Zhang, Shaqiu Huang, Juan Mao, Sai Ou, Xumin Gao, Qun Sun, Di Liu, Yunya Zhang, Ling Yu, Yanling Cheng, Anchun Chen, Shun Front Immunol Immunology Avian Tembusu virus (TMUV) is a novel flavivirus causing severe egg drop and fatal encephalitis in avian in Asia. In the present study, we screened the structural and functional requirements of TMUV capsid protein (CP) for viral morphogenesis using reverse genetics methods in combination with replicon packaging assays. TMUV-CP showed dramatic functional and structural flexibility, and even though 44 residues were removed from the N-terminus, it was still capable of packaging replicon RNA; in addition, 33 residues were deleted from the C-terminus (containing nearly the entire α4-helix), and infectious particles were still produced, although α4-α4’ is supposedly vital for CP dimerization and nucleocapsid formation. We further analyzed two mutants (ΔC20-43 and ΔC64-96 viruses) with relatively large deletions that still replicated well in BHK-21 cells. Our data indicate that internal deletions within CP impaired viral replication or assembly, resulting in attenuated virus proliferation in cells and attenuated virulence in duck embryos, and these deletion mutations are quite stable in cell culture. An in vivo assay indicated that both ΔC20-43 virus and ΔC64-96 virus were highly attenuated in ducklings but still immunogenic. Single-dose immunization with ΔC20-43 virus or ΔC64-96 virus could protect ducklings from a lethal challenge with good antigen clearance. Together, our data shed light on replication/assembly defective TMUV with internal deletions in CP and provide an effective approach to attenuate viral virulence in live vaccines without changing the antigen composition. Frontiers Media S.A. 2021-08-04 /pmc/articles/PMC8371329/ /pubmed/34421904 http://dx.doi.org/10.3389/fimmu.2021.694959 Text en Copyright © 2021 He, Wang, Guo, Mao, Zhang, Hu, Wang, Jia, Zhu, Liu, Zhao, Yang, Wu, Zhang, Huang, Mao, Ou, Gao, Sun, Liu, Zhang, Yu, Cheng and Chen https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
He, Yu
Wang, Xiaoli
Guo, Jiaqi
Mao, Li
Zhang, Senzhao
Hu, Tao
Wang, Mingshu
Jia, Renyong
Zhu, Dekang
Liu, Mafeng
Zhao, Xinxin
Yang, Qiao
Wu, Ying
Zhang, Shaqiu
Huang, Juan
Mao, Sai
Ou, Xumin
Gao, Qun
Sun, Di
Liu, Yunya
Zhang, Ling
Yu, Yanling
Cheng, Anchun
Chen, Shun
Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title_full Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title_fullStr Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title_full_unstemmed Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title_short Replication/Assembly Defective Avian Flavivirus With Internal Deletions in the Capsid Can Be Used as an Approach for Living Attenuated Vaccine
title_sort replication/assembly defective avian flavivirus with internal deletions in the capsid can be used as an approach for living attenuated vaccine
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8371329/
https://www.ncbi.nlm.nih.gov/pubmed/34421904
http://dx.doi.org/10.3389/fimmu.2021.694959
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