Preparation and immunogenicity evaluation of ferritin nanoparticles conjugated with African swine fever virus p30 protein.

Yue ZHANG; Yi RU; Rongzeng HAO; Yang YANG; Longhe ZHAO; Yajun LI; Rui YANG; Bingzhou LU; Haixue ZHENG

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Preparation and immunogenicity evaluation of ferritin nanoparticles conjugated with African swine fever virus p30 protein.

Author: Yue ZHANG ¹ ; Yi RU ¹ ; Rongzeng HAO ¹ ; Yang YANG ¹ ; Longhe ZHAO ¹ ; Yajun LI ² ; Rui YANG ² ; Bingzhou LU ¹ ; Haixue ZHENG ¹
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1. State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, Gansu, China.
2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, Gansu, China.
Publication Type:Journal Article
Keywords: African swine fever; ferritin nanoparticles; immunogenicity evaluation; p30 protein
MeSH: Animals; Nanoparticles/chemistry*; Mice; African Swine Fever Virus/genetics*; Ferritins/chemistry*; Swine; Viral Vaccines/genetics*; African Swine Fever/immunology*; Mice, Inbred BALB C; Viral Proteins/genetics*; Escherichia coli/metabolism*; Dendritic Cells/immunology*; Immunogenicity, Vaccine; Antibodies, Viral/blood*; Female; Capsid Proteins/genetics*
From: Chinese Journal of Biotechnology 2024;40(12):4509-4520
CountryChina
Language:Chinese
Abstract: This study developed ferritin-based nanoparticles carrying the African swine fever virus (ASFV) p30 protein and evaluated their immunogenicity, aiming to provide an experimental basis for the research on nanoparticle vaccines against ASFV. Initially, the gene sequences encoding the p30 protein and SpyTag were fused and inserted into the pCold-I vector to create the pCold-p30 plasmid. The gene sequences encoding SpyCatcher and ferritin were fused and then inserted into the pET-28a(+) vector to produce the pET-F-np plasmid. Both plasmids were expressed in Escherichia coli upon induction. Subsequently, the affinity chromatography-purified p30 protein was conjugated with ferritin in vitro, and the p30-ferritin (F-p30) nanoparticles were purified by size-exclusion chromatography. The morphology and structural integrity of F-p30 nanoparticles were examined by a particle size analyzer and transmission electron microscopy. Mice were immunized with F-p30 nanoparticles, and the humoral and cellular immune responses were assessed. The results showed that F-p30 nanoparticles were successfully prepared, with the particle size of approximately 20 nm. F-p30 nanoparticles were efficiently internalized by bone marrow-derived dendritic cells (BMDCs) cells in vitro. Compared with the p30 protein alone, F-p30 nanoparticles induced elevated levels of specific antibodies and cytokines in mice and stimulated the proliferation of follicular helper T cell (T_FH) and germinal center B cell (GCB) in lymph nodes as well as CD4⁺ and CD8⁺ T cells in the spleen. In conclusion, we successfully prepared F-p30 nanoparticles which significantly enhanced the immunogenicity of p30 protein, giving insights into the development of vaccines against ASFV.