NDLI: Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine

Content Provider	Springer Nature : BioMed Central
Author	Chen, Ya-Yi Yang, Wei-Chen Chang, Yu-Kang Wang, Chi-Young Huang, Wei-Ru Li, Jyun-Yi Chuang, Kuo-Pin Wu, Hung-Yi Chang, Ching-Dong Nielsen, Brent L. Liu, Hung-Jen
Abstract	To increase expression levels of the PCV2 Cap(d41) protein, novel baculovirus surface display vectors with multiple expression cassettes were constructed to create recombinant baculoviruses BacSC-Cap(d41), BacDD-2Cap(d41), BacDD-3Cap(d41), and BacDD-4Cap(d41). Our results reveal that the recombinant baculovirus BacDD-4Cap(d41) was able to express the highest levels of Cap(d41) protein. Optimum conditions for expressing the PCV2 Cap(d41) protein were determined, and our results show that 107 of Sf-9 infected with the recombinant baculovirus BacDD-4Cap(d41) at an MOI of 5 for 3 days showed the highest level of protein expression. Mice immunized with the 4Cap(d41) vaccine which was prepared from the recombinant baculovirus-infected cells (107) elicited higher ELISA titers compared to the Cap (d41) vaccine. The 4Cap(d41) vaccine could elicit anti-PCV2 neutralizing antibodies and IFN-γ in mice, as confirmed by virus neutralization test and IFN-γ ELISA. Moreover, the swine lymphocyte proliferative responses indicated that the 4Cap(d41) vaccine was able to induce a clear cellular immune response. Flow cytometry analysis showed that the percentage of CD4+ T cells and CD4+/CD8+ ratio was increased significantly in SPF pigs immunized with the 4Cap(d41) vaccine. Importantly, the 4Cap(d41) vaccine induced an IFN-γ response, further confirming that its effect is through cellular immunity in SPF pigs. An in vivo challenge study revealed that the 4Cap(d41) and the commercial vaccine groups significantly reduce the viral load of vaccinated pigs as compared with the CE negative control group. Taken together, we have successfully developed a 4Cap(d41) vaccine that may be a potential subunit vaccine for preventing the disease associated with PCV2 infections.
Related Links	https://veterinaryresearch.biomedcentral.com/counter/pdf/10.1186/s13567-020-00836-3.pdf
Ending Page	16
Page Count	16
Starting Page	1
File Format	HTM / HTML
ISSN	12979716
DOI	10.1186/s13567-020-00836-3
Journal	Veterinary Research
Issue Number	1
Volume Number	51
Language	English
Publisher	BioMed Central
Publisher Date	2020-09-09
Access Restriction	Open
Subject Keyword	Veterinary Medicine Veterinary Science Virology Microbiology Immunology PCV2 Cap protein baculovirus surface display vectors 4Cap(d41) vaccine virus neutralization test cellular immune response CD4+ T cells IFN-γ Veterinary Medicine/Veterinary Science
Content Type	Text
Resource Type	Article
Subject	Veterinary Medicine
Journal Impact Factor	3.7/2023
5-Year Journal Impact Factor	3.8/2023

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