Cargando…
Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus
Duck hepatitis A subtype 1 virus (DHAV-1) infection causes high mortality in ducklings, resulting in significant losses to duck industries. VP3 is a structural protein of DHAV-1. However, B-cell epitopes on VP3 have not been investigated. To stimulate VP3 antibody response, eukaryotic expression pla...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589924/ https://www.ncbi.nlm.nih.gov/pubmed/28883462 http://dx.doi.org/10.1038/s41598-017-10909-7 |
_version_ | 1783262436916199424 |
---|---|
author | Wu, Xiaoying Zhang, Tingting Meng, Fanyi Guo, Dongchun Yin, Xiuchen Wulin, Shaozhou Li, Chenxi Zhang, Qingshan Liu, Ming Zhang, Yun |
author_facet | Wu, Xiaoying Zhang, Tingting Meng, Fanyi Guo, Dongchun Yin, Xiuchen Wulin, Shaozhou Li, Chenxi Zhang, Qingshan Liu, Ming Zhang, Yun |
author_sort | Wu, Xiaoying |
collection | PubMed |
description | Duck hepatitis A subtype 1 virus (DHAV-1) infection causes high mortality in ducklings, resulting in significant losses to duck industries. VP3 is a structural protein of DHAV-1. However, B-cell epitopes on VP3 have not been investigated. To stimulate VP3 antibody response, eukaryotic expression plasmid pCI-neo-VP3 was constructed and used as DNA immunogen to prepare mAbs. Western blot showed that 25.5 kDa VP3 could be detected by mAbs in duck embryo fibroblast (DEF) cells transfected with pCI-neo-VP3. Immunofluorescence assay showed that mAbs could specifically bind to DEF cells infected with DHAV-1. DAPI staining indicated that VP3 localizes to the cytoplasm and nucleus of DHAV-1 infected DEF. With neutralizing mAb 3B7, minimal epitope PSNI was mapped. Sequence alignment indicated that (205)PSNI(208) is highly conserved among DHAV-1, but different from those of DHAV-2 and DHAV-3. Epitope peptide reacted specifically with DHAV-1-positive duck sera by dot blotting, revealing PSNI is DHAV-1 type-specific epitope and the importance of these amino acids in antibody-epitope binding reactivity. These findings provided useful information for understanding the antigenicity of VP3 and might be valuable in the development of epitope-based vaccine or diagnostic kit for DHAV-1 infection and provide insights for understanding the pathogenesis of DHAV-1. |
format | Online Article Text |
id | pubmed-5589924 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-55899242017-09-13 Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus Wu, Xiaoying Zhang, Tingting Meng, Fanyi Guo, Dongchun Yin, Xiuchen Wulin, Shaozhou Li, Chenxi Zhang, Qingshan Liu, Ming Zhang, Yun Sci Rep Article Duck hepatitis A subtype 1 virus (DHAV-1) infection causes high mortality in ducklings, resulting in significant losses to duck industries. VP3 is a structural protein of DHAV-1. However, B-cell epitopes on VP3 have not been investigated. To stimulate VP3 antibody response, eukaryotic expression plasmid pCI-neo-VP3 was constructed and used as DNA immunogen to prepare mAbs. Western blot showed that 25.5 kDa VP3 could be detected by mAbs in duck embryo fibroblast (DEF) cells transfected with pCI-neo-VP3. Immunofluorescence assay showed that mAbs could specifically bind to DEF cells infected with DHAV-1. DAPI staining indicated that VP3 localizes to the cytoplasm and nucleus of DHAV-1 infected DEF. With neutralizing mAb 3B7, minimal epitope PSNI was mapped. Sequence alignment indicated that (205)PSNI(208) is highly conserved among DHAV-1, but different from those of DHAV-2 and DHAV-3. Epitope peptide reacted specifically with DHAV-1-positive duck sera by dot blotting, revealing PSNI is DHAV-1 type-specific epitope and the importance of these amino acids in antibody-epitope binding reactivity. These findings provided useful information for understanding the antigenicity of VP3 and might be valuable in the development of epitope-based vaccine or diagnostic kit for DHAV-1 infection and provide insights for understanding the pathogenesis of DHAV-1. Nature Publishing Group UK 2017-09-07 /pmc/articles/PMC5589924/ /pubmed/28883462 http://dx.doi.org/10.1038/s41598-017-10909-7 Text en © The Author(s) 2017 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/. |
spellingShingle | Article Wu, Xiaoying Zhang, Tingting Meng, Fanyi Guo, Dongchun Yin, Xiuchen Wulin, Shaozhou Li, Chenxi Zhang, Qingshan Liu, Ming Zhang, Yun Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title | Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title_full | Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title_fullStr | Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title_full_unstemmed | Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title_short | Mapping a Type-specific Epitope by Monoclonal Antibody against VP3 Protein of Duck Hepatitis A Type 1 Virus |
title_sort | mapping a type-specific epitope by monoclonal antibody against vp3 protein of duck hepatitis a type 1 virus |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589924/ https://www.ncbi.nlm.nih.gov/pubmed/28883462 http://dx.doi.org/10.1038/s41598-017-10909-7 |
work_keys_str_mv | AT wuxiaoying mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT zhangtingting mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT mengfanyi mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT guodongchun mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT yinxiuchen mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT wulinshaozhou mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT lichenxi mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT zhangqingshan mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT liuming mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus AT zhangyun mappingatypespecificepitopebymonoclonalantibodyagainstvp3proteinofduckhepatitisatype1virus |