FliC蛋白佐剂对伪狂犬病病毒Bartha-K61株弱毒疫苗的黏膜免疫作用

doi:10.16431/j.cnki.1671-7236.2025.10.031

Abstract

Abstract: 【Objective】 This experiment aimed to investigate the immunopotentiating effect of FliC protein as a mucosal adjuvant on the Pseudorabies virus (PRV) Bartha-K61 attenuated vaccine strain,so as to lay a foundation for developing PRV-targeted mucosal vaccines.【Method】 FliC gene was cloned from Salmonella Typhimurium and linked to the prokaryotic expression vector pET-28a(+)，the linked product was transformed into Escherichia coli (E.coli) Trans10 competent cells,and the plasmid was extracted for double enzyme digestion identification and sequencing.FliC-pET-28a was transformed into E.coli BL21(DE3) competent cells to induce the expression of FliC protein,and the purified FliC protein was identified by Coomassie brilliant blue staining and Western blotting.Bartha-K61 virus suspension group and the commercial Bartha-K61 vaccine group were set up as control.FliC protein as mucosal adjuvant was mixed with Bartha-K61 virus solution,BALB/c mice were intranasally immunized twice.The levels of IgG and IgA in the serum of mice before immunization (day 0) and at post-immunization days 7,14,21,28 and 35 were measured.Mice in each group were challenged with PRV GD-WH of 10^4.5 TCID₅₀ on the 35th day after immunization and the survival was observed and recorded daily after challenge.【Result】 The recombinant plasmid FliC-pET-28a was successfully obtained,the soluble expression of FliC was realized,and the size of the purified FliC protein was approximately 58 ku.The results of mouse immune test showed that the serum IgG and IgA antibodies of Bartha-K61 virus solution+FliC protein group were significantly higher than those of Bartha-K61 virus solution group and commercial Bartha-K61 vaccine group (P＜0.05).The results of challenge protection test showed that Bartha-K61 virus solution+FliC protein could provide 100% protection to mice against the lethal attack of PRV GD-WH,which was higher than that of Bartha-K61 virus solution alone.【Conclusion】 FliC protein adjuvant could enhance mucosal immunity of PRV Bartha-K61 attenuated vaccine strain,and enable mice to obtain certain immune protection.

Key words: FliC protein; Pseudorabies virus attenuated vaccine; mucosal immunity

CLC Number:

LIANG Lirong, KE Haiyi, ZANG Yingan, ZHAI Shaolun, KANG Huahua, JIANG Zhiyong, LI Yan, ZHANG Kunli, ZHOU Xia, GOU Hongchao, LI Chunling. Effect of FliC Protein Adjuvant on Mucosal Immunity of Pseudorabies Virus Bartha-K61 Attenuated Vaccine[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4864-4873.

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Effect of FliC Protein Adjuvant on Mucosal Immunity of Pseudorabies Virus Bartha-K61 Attenuated Vaccine

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