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Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development
Live-attenuated vaccines (LAVs) represent a promising approach for flavivirus vaccine development. In the present study, we demonstrated a method for generating flavivirus LAVs based on breaking spatially and temporally regulated C-prM cleavage to disturb the viral assembly process, using an avian f...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9098475/ https://www.ncbi.nlm.nih.gov/pubmed/35550523 http://dx.doi.org/10.1038/s41541-022-00468-y |
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author | He, Yu Guo, Jiaqi Wang, Xiaoli Zhang, Senzhao Mao, Li 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 Cheng, Anchun Chen, Shun |
author_facet | He, Yu Guo, Jiaqi Wang, Xiaoli Zhang, Senzhao Mao, Li 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 Cheng, Anchun Chen, Shun |
author_sort | He, Yu |
collection | PubMed |
description | Live-attenuated vaccines (LAVs) represent a promising approach for flavivirus vaccine development. In the present study, we demonstrated a method for generating flavivirus LAVs based on breaking spatially and temporally regulated C-prM cleavage to disturb the viral assembly process, using an avian flavivirus (Tembusu virus) as the model. Using reverse genetics technology, we successfully generated two recombinant viruses (CQW1-IRES-mC and CQW1-MINI-mC) with bicistronic genomic RNA in which native capsid genes were deleted and instead expressed in the 3’UTR under the control of an internal ribosome entry site (IRES) or minimum IRES. Both viruses showed a significantly attenuated phenotype in vitro due to impaired viral assembly, and the engineered mutations were genetically stable in vitro within ten passages. Importantly, their virulence was also highly attenuated in ducklings and suckling mice and did not cause any overt clinical symptoms or mortality. In addition, a single dose of immunization with any of these mutant viruses could completely protect ducklings from a lethal challenge, and no viremia was detected after immunization and challenge, even though the viruses induced a relatively moderate immune response in terms of the T-lymphocytes proliferative response and the level of neutralization antibodies compared with that obtained with the wild-type virus. Besides, a recombinant virus ectopically expressing the prM-E protein was also generated in the present study, but this virus was too attenuated with severely decreased proliferation. Our results indicated that the use of a recombinant flavivirus that ectopically expresses structural proteins could be an effective and universal method for flavivirus LAVs development. |
format | Online Article Text |
id | pubmed-9098475 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-90984752022-05-14 Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development He, Yu Guo, Jiaqi Wang, Xiaoli Zhang, Senzhao Mao, Li 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 Cheng, Anchun Chen, Shun NPJ Vaccines Article Live-attenuated vaccines (LAVs) represent a promising approach for flavivirus vaccine development. In the present study, we demonstrated a method for generating flavivirus LAVs based on breaking spatially and temporally regulated C-prM cleavage to disturb the viral assembly process, using an avian flavivirus (Tembusu virus) as the model. Using reverse genetics technology, we successfully generated two recombinant viruses (CQW1-IRES-mC and CQW1-MINI-mC) with bicistronic genomic RNA in which native capsid genes were deleted and instead expressed in the 3’UTR under the control of an internal ribosome entry site (IRES) or minimum IRES. Both viruses showed a significantly attenuated phenotype in vitro due to impaired viral assembly, and the engineered mutations were genetically stable in vitro within ten passages. Importantly, their virulence was also highly attenuated in ducklings and suckling mice and did not cause any overt clinical symptoms or mortality. In addition, a single dose of immunization with any of these mutant viruses could completely protect ducklings from a lethal challenge, and no viremia was detected after immunization and challenge, even though the viruses induced a relatively moderate immune response in terms of the T-lymphocytes proliferative response and the level of neutralization antibodies compared with that obtained with the wild-type virus. Besides, a recombinant virus ectopically expressing the prM-E protein was also generated in the present study, but this virus was too attenuated with severely decreased proliferation. Our results indicated that the use of a recombinant flavivirus that ectopically expresses structural proteins could be an effective and universal method for flavivirus LAVs development. Nature Publishing Group UK 2022-05-12 /pmc/articles/PMC9098475/ /pubmed/35550523 http://dx.doi.org/10.1038/s41541-022-00468-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article He, Yu Guo, Jiaqi Wang, Xiaoli Zhang, Senzhao Mao, Li 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 Cheng, Anchun Chen, Shun Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title | Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title_full | Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title_fullStr | Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title_full_unstemmed | Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title_short | Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
title_sort | assembly-defective tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9098475/ https://www.ncbi.nlm.nih.gov/pubmed/35550523 http://dx.doi.org/10.1038/s41541-022-00468-y |
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