Construction and immune efficacy evaluation of BNeV VLPs based on VP1 protein in mice
10.16303/j.cnki.1005-4545.2025.03.03
- VernacularTitle:牛纽布病毒病毒样颗粒的制备及对小鼠的免疫效果评价
- Author:
Lu DING
1
;
Xiangyue HUANG
;
Jinbo WU
;
Chaohui ZHANG
;
Qing ZHU
;
Chenxi ZHU
;
Gu-nan DENG
;
Ajia AKE
;
Chunsai HE
;
Yuanzhen MA
;
Bin ZHANG
Author Information
1. 西南民族大学畜牧兽医学院,四川成都 610041
- Publication Type:Journal Article
- Keywords:
bovine nebovirus;
VP1 protein;
virus-like particles;
immunogenicity;
mice
- From:
Chinese Journal of Veterinary Science
2025;45(3):412-419
- CountryChina
- Language:Chinese
-
Abstract:
The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4％.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.
