Immunogenicity and protective effect of recombinant Brucella abortus Ndk (rNdk) against a virulent strain B. abortus 544 infection in BALB/c mice.
In this study, we particularly evaluated the protective effect of recombinant protein encoded by Brucella abortus 544 ndk (nucleoside diphosphate kinase) gene against B. abortus infection in the BALB/c mice. Cloning and expression of B. abortus Ndk was accomplished by PCR amplification into a pMAL expression system, and purification of a recombinant Ndk (rNdk). As for the determination of IgG responses, rNdk induced vigorous IgG production, especially higher in IgG2a compared to IgG1 with titers of 5.2 and 4.8, respectively, whereas titers of these in mice immunized with MBP were 2.4 of IgG2a and 2.6 of IgG1. The analysis of cytokine has revealed that rNdk can strongly induce production of IFN-γ as well as proinflammatory cytokines (TNF, MCP1 and IL-6) but not much IL-10, suggesting rNdk elicited predominantly cell-mediated immune responses. Furthermore, the spleen proliferation and bacterial burden in the spleen of rNdk immunized mice were significantly lower than those of MBP-immunized mice against virulent B. abortus challenge (P < 0.01). Conclusionly, rNdk immunization enables to elicit both of the humoral and cellular response, ultimately enhancing protection level in experimental mice, suggesting that rNdk of B. abortus might be a useful candidate for subunit vaccine for brucellosis in animals.
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
Huynh Tan
HT
Hop
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
Hannah Leah
HL
Simborio
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
Alisha Wehdnesday Bernardo
AW
Reyes
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
Lauren Togonon
LT
Arayan
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
WonGi
W
Min
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea.
Hu Jang
HJ
Lee
School of Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Dong Hee
DH
Kim
Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Hong Hee
HH
Chang
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Korea Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea kimsuk@gnu.ac.kr.
Suk
S
Kim
D000818:Animals; D002003:Brucella abortus / Q000235:genetics* / Q000276:immunology*; D002006:Brucellosis / Q000276:immunology* / Q000517:prevention & control; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism; D022223:Vaccines, Subunit
D007074:Immunoglobulin G; D015850:Interleukin-6; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D022223:Vaccines, Subunit; D016753:Interleukin-10; D007371:Interferon-gamma; D009701:Nucleoside-Diphosphate Kinase
Brucella abortus; nucleoside diphosphate kinase (Ndk); protection; vaccine