title: Vaccination with recombinant flagellar proteins FlgJ and FliN induce protection against Brucella abortus 544 infection in BALB/c mice.
abstract: Brucella has been considered as a non-motile, facultative intracellular pathogenic bacterium. However, the genome sequences of different Brucella species reveal the presence of the flagellar genes needed for the construction of a functional flagellum. Due to its roles in the interaction between pathogen and host, we hypothesized that some of the flagellar proteins might induce protective immune responses and these proteins will be good subunit vaccine candidates. This study was conducted to screening of protective antigens among these flagellar proteins. Firstly, according to the putative functional roles, a total of 30 flagellar genes of Brucella abortus were selected for in vitro expression. 15 of these flagellar genes were successfully expressed as his-tagged recombinant proteins in Escherichia coli ER2566. Then, these proteins were purified and used to analyze their T cell immunity induction activity by an in vitro gamma interferon (IFN-γ) assay. Five of the flagellar proteins could stimulate significantly higher levels of IFN-γ secretion in splenocytes from S19 immunized mice, indicating their T cell induction activity. Finally, immunogenicity and protection activity of these 5 flagellar proteins were evaluated in BALB/c mice. Results showed that immunization with FlgJ (BAB1_0260) or FliN (BAB2_0122) plus adjuvant could provide protection against B. abortus 544 infection. Furthermore, mice immunized with FlgJ and FliN developed a vigorous immunoglobulin G response, and in vitro stimulation of their splenocytes with immunizing proteins induced the secretion of IFN-γ. Altogether, these data suggest that flagellar proteins FlgJ and FliN are protective antigens that could produce humoral and cell-mediated responses in mice and candidates for use in future studies of vaccination against brucellosis.
authors.affiliation: College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Xianbo
authors.initials: X
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Xu
authors.forename: Yongfei
authors.initials: Y
authors.lastname: Xie
authors.forename: Yefeng
authors.initials: Y
authors.lastname: Qiu
authors.forename: Simei
authors.initials: S
authors.lastname: Fu
authors.forename: Xitong
authors.initials: X
authors.lastname: Yuan
authors.forename: Yuehua
authors.initials: Y
authors.lastname: Ke
authors.forename: Shuang
authors.initials: S
authors.lastname: Yu
authors.forename: Xinying
authors.initials: X
authors.lastname: Du
authors.forename: Mingquan
authors.initials: M
authors.lastname: Cui
authors.forename: Yanfen
authors.initials: Y
authors.lastname: Chen
authors.forename: Tongkun
authors.initials: T
authors.lastname: Wang
authors.forename: Zhoujia
authors.initials: Z
authors.lastname: Wang
authors.forename: Yaqing
authors.initials: Y
authors.lastname: Yu
authors.forename: Kehe
authors.initials: K
authors.lastname: Huang
authors.forename: Liuyu
authors.initials: L
authors.lastname: Huang
authors.forename: Guangneng
authors.initials: G
authors.lastname: Peng
authors.forename: Zeliang
authors.initials: Z
authors.lastname: Chen
authors.forename: Yufei
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology; D002004:Brucella Vaccine / Q000008:administration & dosage / Q000276:immunology*; D002003:Brucella abortus / Q000276:immunology*; D002006:Brucellosis / Q000276:immunology / Q000517:prevention & control*; D005260:Female; D056724:Immunity, Humoral / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology; D013997:Time Factors; D014611:Vaccination; D022223:Vaccines, Subunit / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D002004:Brucella Vaccine; C077389:FliN protein, Bacteria; D011994:Recombinant Proteins; D022223:Vaccines, Subunit