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Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells

Newcastle disease (ND) is an acute and highly contagious infectious disease found in poultry. Although commercial ND virus (NDV) vaccines are universally used, some case reports persistently documented vaccination failure. Therefore, novel strategies are still required to control the occurrence of t...

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Autores principales: Fan, Wenqi, Ji, Pinpin, Sun, Xuwen, Kong, Min, Zhou, Ning, Zhang, Qiang, Wang, Ying, Liu, Qianqian, Li, Xiaoxuan, Zhou, En-Min, Zhao, Qin, Sun, Yani
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9426676/
https://www.ncbi.nlm.nih.gov/pubmed/36051768
http://dx.doi.org/10.3389/fmicb.2022.956561
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author Fan, Wenqi
Ji, Pinpin
Sun, Xuwen
Kong, Min
Zhou, Ning
Zhang, Qiang
Wang, Ying
Liu, Qianqian
Li, Xiaoxuan
Zhou, En-Min
Zhao, Qin
Sun, Yani
author_facet Fan, Wenqi
Ji, Pinpin
Sun, Xuwen
Kong, Min
Zhou, Ning
Zhang, Qiang
Wang, Ying
Liu, Qianqian
Li, Xiaoxuan
Zhou, En-Min
Zhao, Qin
Sun, Yani
author_sort Fan, Wenqi
collection PubMed
description Newcastle disease (ND) is an acute and highly contagious infectious disease found in poultry. Although commercial ND virus (NDV) vaccines are universally used, some case reports persistently documented vaccination failure. Therefore, novel strategies are still required to control the occurrence of the disease in chickens. Recently, nanobodies (Nbs), which have the advantages of small molecular weight and low production costs, have been shown to be promising therapeutics against viral infection. In the present study, a total of 16 Nbs against NDV nucleocapsid protein (NP) were screened from two libraries against NDV using phage display technology. Of the 16 screened Nbs, eight were prevented from binding to NDV NP protein through administering positive chicken sera for anti-NDV antibodies, indicating that the epitopes recognized by these eight Nbs were able to induce the immune response after the chickens were infected with NDV stock. Subsequently, transfection assay, construction of recombinant DF-1 cells capable of expressing different nanobodies and viral inhibition assay were used to screen the nanobodies inhibiting NDV replication. The results demonstrated that Nb18, Nb30, and Nb88 significantly inhibited the replication of Class I and different genotypes of Class II NDV strains in DF-1 cells when they were expressed in the cytoplasm. Collectively, these nanobodies provided new tools for researching the functions of NDV NP protein and may be used as a novel strategy for designing drugs against NDV infection in chickens.
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spelling pubmed-94266762022-08-31 Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells Fan, Wenqi Ji, Pinpin Sun, Xuwen Kong, Min Zhou, Ning Zhang, Qiang Wang, Ying Liu, Qianqian Li, Xiaoxuan Zhou, En-Min Zhao, Qin Sun, Yani Front Microbiol Microbiology Newcastle disease (ND) is an acute and highly contagious infectious disease found in poultry. Although commercial ND virus (NDV) vaccines are universally used, some case reports persistently documented vaccination failure. Therefore, novel strategies are still required to control the occurrence of the disease in chickens. Recently, nanobodies (Nbs), which have the advantages of small molecular weight and low production costs, have been shown to be promising therapeutics against viral infection. In the present study, a total of 16 Nbs against NDV nucleocapsid protein (NP) were screened from two libraries against NDV using phage display technology. Of the 16 screened Nbs, eight were prevented from binding to NDV NP protein through administering positive chicken sera for anti-NDV antibodies, indicating that the epitopes recognized by these eight Nbs were able to induce the immune response after the chickens were infected with NDV stock. Subsequently, transfection assay, construction of recombinant DF-1 cells capable of expressing different nanobodies and viral inhibition assay were used to screen the nanobodies inhibiting NDV replication. The results demonstrated that Nb18, Nb30, and Nb88 significantly inhibited the replication of Class I and different genotypes of Class II NDV strains in DF-1 cells when they were expressed in the cytoplasm. Collectively, these nanobodies provided new tools for researching the functions of NDV NP protein and may be used as a novel strategy for designing drugs against NDV infection in chickens. Frontiers Media S.A. 2022-07-27 /pmc/articles/PMC9426676/ /pubmed/36051768 http://dx.doi.org/10.3389/fmicb.2022.956561 Text en Copyright © 2022 Fan, Ji, Sun, Kong, Zhou, Zhang, Wang, Liu, Li, Zhou, Zhao and Sun. 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 Microbiology
Fan, Wenqi
Ji, Pinpin
Sun, Xuwen
Kong, Min
Zhou, Ning
Zhang, Qiang
Wang, Ying
Liu, Qianqian
Li, Xiaoxuan
Zhou, En-Min
Zhao, Qin
Sun, Yani
Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title_full Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title_fullStr Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title_full_unstemmed Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title_short Screening and identification of nucleocapsid protein-nanobodies that inhibited Newcastle disease virus replication in DF-1 cells
title_sort screening and identification of nucleocapsid protein-nanobodies that inhibited newcastle disease virus replication in df-1 cells
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9426676/
https://www.ncbi.nlm.nih.gov/pubmed/36051768
http://dx.doi.org/10.3389/fmicb.2022.956561
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