A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice
Developing new vaccine candidates is considered the best strategy for protecting poultry against artificial haemagglutinin (A/H5N1) strains. The transient expression system in plants has been a very efficient method for rapidly producing haemagglutinin-based recombinant vaccines. In this study, two...
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| Format: | Article |
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MDPI AG
2019-10-01
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| Series: | Applied Sciences |
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| Online Access: | https://www.mdpi.com/2076-3417/9/21/4605 |
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| author | Van Thi Pham Thuong Thi Ho Hoang Trong Phan Thanh Hoa Le Ngoc Bich Pham Udo Conrad Trang Huyen Vu Ha Hoang Chu |
| author_facet | Van Thi Pham Thuong Thi Ho Hoang Trong Phan Thanh Hoa Le Ngoc Bich Pham Udo Conrad Trang Huyen Vu Ha Hoang Chu |
| author_sort | Van Thi Pham |
| collection | DOAJ |
| description | Developing new vaccine candidates is considered the best strategy for protecting poultry against artificial haemagglutinin (A/H5N1) strains. The transient expression system in plants has been a very efficient method for rapidly producing haemagglutinin-based recombinant vaccines. In this study, two novel artificial trimeric haemagglutinin constructs representing A/H5N1 strains that were detected in poultry from 2005 to 2015 in Vietnam, H5.c1 (representing all of the subclades 1.1, 1.1.1, and 1.1.2) and H5.c2 (representing all of the subclades 2.3.2.1, 2.3.2.1a, 2.3.2.1b, and 2.3.2.1c), were designed for transient expression in <i>Nicotiana benthamiana</i> via agroinfiltration. However, only the H5.c1 protein, which showed the best expression and biofunction via the haemagglutination test, was selected for purification by immobilized metal ion affinity chromatography (IMAC). The trimeric structure of the IMAC-purified H5.c1 protein was well characterized by cross-linking reaction and size exclusion chromatography. An indirect ELISA and Western blot analysis of vaccinated mouse sera demonstrated that the H5.c1 protein strongly induced HA-specific Immunoglobulin G (IgG) immune responses. Notably, the H5.c1 protein induced strongly neutralizing antibodies against homologous H5.c1 protein and that of three heterologous native strains of clade, 1, 1.1, and 2.3.2.1c, in haemagglutination inhibition assays. Therefore, the plant-based artificial H5.c1 protein can be a promising vaccine candidate for conferring poultry resistance against A/H5N1 viruses in Vietnam. |
| first_indexed | 2024-12-21T21:47:08Z |
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| id | doaj.art-970e41687c1440c2ac49775ba1702e18 |
| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-12-21T21:47:08Z |
| publishDate | 2019-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-970e41687c1440c2ac49775ba1702e182022-12-21T18:49:11ZengMDPI AGApplied Sciences2076-34172019-10-01921460510.3390/app9214605app9214605A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in MiceVan Thi Pham0Thuong Thi Ho1Hoang Trong Phan2Thanh Hoa Le3Ngoc Bich Pham4Udo Conrad5Trang Huyen Vu6Ha Hoang Chu7Institute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Plant Genetics and Crop Plant Research (IPK), 06466 Gatersleben, GermanyInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamInstitute of Biotechnology (IBT), Vietnam Academy of Science and Technology (VAST), Hanoi 100000, VietnamDeveloping new vaccine candidates is considered the best strategy for protecting poultry against artificial haemagglutinin (A/H5N1) strains. The transient expression system in plants has been a very efficient method for rapidly producing haemagglutinin-based recombinant vaccines. In this study, two novel artificial trimeric haemagglutinin constructs representing A/H5N1 strains that were detected in poultry from 2005 to 2015 in Vietnam, H5.c1 (representing all of the subclades 1.1, 1.1.1, and 1.1.2) and H5.c2 (representing all of the subclades 2.3.2.1, 2.3.2.1a, 2.3.2.1b, and 2.3.2.1c), were designed for transient expression in <i>Nicotiana benthamiana</i> via agroinfiltration. However, only the H5.c1 protein, which showed the best expression and biofunction via the haemagglutination test, was selected for purification by immobilized metal ion affinity chromatography (IMAC). The trimeric structure of the IMAC-purified H5.c1 protein was well characterized by cross-linking reaction and size exclusion chromatography. An indirect ELISA and Western blot analysis of vaccinated mouse sera demonstrated that the H5.c1 protein strongly induced HA-specific Immunoglobulin G (IgG) immune responses. Notably, the H5.c1 protein induced strongly neutralizing antibodies against homologous H5.c1 protein and that of three heterologous native strains of clade, 1, 1.1, and 2.3.2.1c, in haemagglutination inhibition assays. Therefore, the plant-based artificial H5.c1 protein can be a promising vaccine candidate for conferring poultry resistance against A/H5N1 viruses in Vietnam.https://www.mdpi.com/2076-3417/9/21/4605a/h5n1haemagglutinintrimerization<i>nicotiana benthamiana</i>agroinfiltrationneutralizing antibody |
| spellingShingle | Van Thi Pham Thuong Thi Ho Hoang Trong Phan Thanh Hoa Le Ngoc Bich Pham Udo Conrad Trang Huyen Vu Ha Hoang Chu A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice Applied Sciences a/h5n1 haemagglutinin trimerization <i>nicotiana benthamiana</i> agroinfiltration neutralizing antibody |
| title | A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice |
| title_full | A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice |
| title_fullStr | A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice |
| title_full_unstemmed | A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice |
| title_short | A Plant-Based Artificial Haemagglutinin (A/H5N1) Strongly Induced Neutralizing Immune Responses in Mice |
| title_sort | plant based artificial haemagglutinin a h5n1 strongly induced neutralizing immune responses in mice |
| topic | a/h5n1 haemagglutinin trimerization <i>nicotiana benthamiana</i> agroinfiltration neutralizing antibody |
| url | https://www.mdpi.com/2076-3417/9/21/4605 |
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