Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development
Abstract Both current live, attenuated, and killed virus vaccines for bovine viral diarrhea virus (BVDV) have their limitations. Here, we report the development of a BVDV subunit vaccine by (i) the expression of a secreted form of a recombinant E2 glycoprotein using BHK21 cells and (ii) determinatio...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-26766-y |
| _version_ | 1797958756962664448 |
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| author | Yi Ting Lo Martin D. Ryan Garry A. Luke Wan Chen Chang Hsing Chieh Wu |
| author_facet | Yi Ting Lo Martin D. Ryan Garry A. Luke Wan Chen Chang Hsing Chieh Wu |
| author_sort | Yi Ting Lo |
| collection | DOAJ |
| description | Abstract Both current live, attenuated, and killed virus vaccines for bovine viral diarrhea virus (BVDV) have their limitations. Here, we report the development of a BVDV subunit vaccine by (i) the expression of a secreted form of a recombinant E2 glycoprotein using BHK21 cells and (ii) determination of the immune responses in mice. The E2 glycoprotein was modified by deletion of the C-terminal transmembrane anchor domain and fusion to a V5 epitope tag. This allowed detection using anti-V5 monoclonal antibodies together with simple purification of the expressed, secreted, form of E2 from the cell media. Furthermore, we genetically fused green fluorescent protein (GFP) linked to E2 via a Thosea asigna virus 2A (T2A) ribosome skipping sequence thereby creating a self-processing polyprotein [GFP-T2A-BVDV-E2trunk-V5], producing discrete [GFP-T2A] and [E2trunk-V5] translation products: GFP fluorescence acts, therefore, as a surrogate marker of E2 expression, BALB/c mice were inoculated with [E2trunk-V5] purified from cell media and both humoral and cellular immune responses were observed. Our antigen expression system provides, therefore, both (i) a simple antigen purification protocol together with (ii) a feasible strategy for further, large-scale, production of vaccines. |
| first_indexed | 2024-04-11T00:23:14Z |
| format | Article |
| id | doaj.art-addde191cd7b4c3386675eb11c6f12b4 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T00:23:14Z |
| publishDate | 2023-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-addde191cd7b4c3386675eb11c6f12b42023-01-08T12:10:46ZengNature PortfolioScientific Reports2045-23222023-01-0113111110.1038/s41598-022-26766-yImmunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine developmentYi Ting Lo0Martin D. Ryan1Garry A. Luke2Wan Chen Chang3Hsing Chieh Wu4International Degree Program in Animal Vaccine Technology, International College, National Pingtung University of Science and TechnologyBiomedical Sciences Research Complex, School of Biology, University of St AndrewsBiomedical Sciences Research Complex, School of Biology, University of St AndrewsInternational Degree Program in Animal Vaccine Technology, International College, National Pingtung University of Science and TechnologyInternational Degree Program in Animal Vaccine Technology, International College, National Pingtung University of Science and TechnologyAbstract Both current live, attenuated, and killed virus vaccines for bovine viral diarrhea virus (BVDV) have their limitations. Here, we report the development of a BVDV subunit vaccine by (i) the expression of a secreted form of a recombinant E2 glycoprotein using BHK21 cells and (ii) determination of the immune responses in mice. The E2 glycoprotein was modified by deletion of the C-terminal transmembrane anchor domain and fusion to a V5 epitope tag. This allowed detection using anti-V5 monoclonal antibodies together with simple purification of the expressed, secreted, form of E2 from the cell media. Furthermore, we genetically fused green fluorescent protein (GFP) linked to E2 via a Thosea asigna virus 2A (T2A) ribosome skipping sequence thereby creating a self-processing polyprotein [GFP-T2A-BVDV-E2trunk-V5], producing discrete [GFP-T2A] and [E2trunk-V5] translation products: GFP fluorescence acts, therefore, as a surrogate marker of E2 expression, BALB/c mice were inoculated with [E2trunk-V5] purified from cell media and both humoral and cellular immune responses were observed. Our antigen expression system provides, therefore, both (i) a simple antigen purification protocol together with (ii) a feasible strategy for further, large-scale, production of vaccines.https://doi.org/10.1038/s41598-022-26766-y |
| spellingShingle | Yi Ting Lo Martin D. Ryan Garry A. Luke Wan Chen Chang Hsing Chieh Wu Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development Scientific Reports |
| title | Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development |
| title_full | Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development |
| title_fullStr | Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development |
| title_full_unstemmed | Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development |
| title_short | Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development |
| title_sort | immunogenicity of a secreted c terminally truncated form of bovine viral diarrhea virus e2 glycoprotein as a potential candidate in subunit vaccine development |
| url | https://doi.org/10.1038/s41598-022-26766-y |
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