Preparation of Chicken Anemia Virus (CAV) Virus-Like Particles and Chicken Interleukin-12 for Vaccine Development Using a Baculovirus Expression System
| Autor: | Ta-Yuan Tseng, Yu-Chen Hsu, Jui-Hung Shien, Ming-Kun Hsieh, Shan-Chia Ou, Yee-Chen Liu, Poa-Chun Chang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Microbiology (medical)
musculoskeletal diseases animal structures viruses Sf9 Article Virus law.invention virus-like particle Virus-like particle adjuvant Serial passage Interferon law medicine Immunology and Allergy chicken infectious anemia vaccine Molecular Biology chicken IL-12 General Immunology and Microbiology biology chicken anemia virus medicine.disease Virology Infectious Diseases vaccine development embryonic structures biology.protein Poultry disease Recombinant DNA Antibody medicine.drug |
| Zdroj: | Pathogens Volume 8 Issue 4 |
| ISSN: | 2076-0817 |
| DOI: | 10.3390/pathogens8040262 |
| Popis: | Chicken infectious anemia (CIA) is a poultry disease that causes huge economic losses in the poultry industry worldwide. Commercially available CIA vaccines are derived from wild-type chicken anemia viruses (CAVs) by serial passage in cells or chicken embryos. However, these vaccinal viruses are not completely attenuated therefore, they can be transmitted vertically and horizontally, and may induce clinical symptoms in young birds. In this study, we sought to eliminate these issues by developing a subunit vaccine exploiting the CAV structural proteins, engineering recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells that contained both the viral protein 1 (VP1) and VP2 of CAV. Moreover, we produced single-chain chicken interleukin-12 (chIL-12) in the same system, to serve as an adjuvant. The recombinant VP1 was recognized by chicken anti-CAV polyclonal antibodies in Western blotting and immunofluorescence assays, and the bioactivity of the recombinant chIL-12 was confirmed by stimulating interferon-&gamma (IFN-&gamma ) secretion in chicken splenocytes. Furthermore, the ability of the recombinant VP1 to generate self-assembling virus-like particles (VLPs) was confirmed by transmission electron microscopy. Specific pathogen-free (SPF) chickens inoculated with VLPs and co-administered the recombinant chIL-12 induced high CAV-specific antibodies and cell-mediated immunity. Taken together, the VLPs produced by the baculovirus expression system have the potential to be a safe and effective CIA vaccine. Finally, we demonstrated the utility of recombinant chIL-12 as an adjuvant for poultry vaccine development. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|