Subunit vaccine of PCV3 capsid protein produced by sf9 cells with double knockout of Caspase-1 and Dronc induces strong immune response in mice

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology Pub Date : 2025-03-04 DOI:10.1016/j.vetmic.2025.110452

Shuo Li , Ruihong Guo , Yinxiang Fang , Chunhong Zhang , Linyu Jiang , Weixin Jia , Zhangyong Ning

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引用次数: 0

Abstract

Porcine circovirus type 3 (PCV3) associated with multisystemic clinicopathological diseases in swine herds has caused economic losses and there is no available commercial vaccine. Production of PCV3 capsid protein (Cap) by Spodoptera frugiperda 9 (sf9) cells using baculovirus expression vector system (BEVS) is a valid strategy to develop vaccines. Here, we report that subunit vaccine of PCV3 produced by sf9 cells with double knockout of Caspase-1 and Dronc genes induces strong immune response in mice. Three kinds of knockout sf9 cells aimed at Caspase-1 gene, Dronc gene and both genes were successfully generated by clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR-Cas9) system, and sequence analysis confirmed this. The anti-apoptosis ability of three kinds of knockout sf9 cells was assessed, and double knockout sf9 cells are the best. The expression of PCV3 Cap was enhanced in double knockout sf9 cells compared to wild type sf9 cells, and subunit vaccines were produced by PCV3 Cap expressed from double knockout sf9 cells and wild type cells, respectively. Results of immunological experiment in mice showed subunit vaccine of PCV3 Cap from double knockout sf9 cells induces higher level of serum antibody, stimulates lymphocyte proliferation and enhances expression of IL-2, IFN-γ, IL-4 and IL-10 compared to wild type cells. These results present knockout sf9 cells to enhance the expression of protein in BEVS, and provide a technical platform for vaccine development of PCV3.

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来源期刊

Veterinary microbiology 农林科学-兽医学

CiteScore

5.90

自引率

6.10%

发文量

221

审稿时长

52 days

期刊介绍： Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal. Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge. Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.