Effects of partial deletion of the wzm and wzt genes on lipopolysaccharide synthesis and virulence of Brucella abortus S19

Wang,Xiuran; Wang,Lin; Lu,Tiancheng; Yang,Yanling; Chen,Si; Zhang,Rui; Lang,Xulong; Yan,Guangmou; Qian,Jing; Wang,Xiaoxu; Meng,Lingyi; Wang,Xinglong

doi:10.3892/mmr.2014.2104

Effects of partial deletion of the wzm and wzt genes on lipopolysaccharide synthesis and virulence of Brucella abortus S19

Authors:
- Xiuran Wang
- Lin Wang
- Tiancheng Lu
- Yanling Yang
- Si Chen
- Rui Zhang
- Xulong Lang
- Guangmou Yan
- Jing Qian
- Xiaoxu Wang
- Lingyi Meng
- Xinglong Wang
View Affiliations

Affiliations: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, AMMS, Changchun, Jilin 130122, P.R. China, Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin 130062, P.R. China, School of Life Science, Jilin Agricultural University, Changchun, Jilin 130118, P.R. China, State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun, Jilin 130122, P.R. China, Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, AMMS, Changchun, Jilin 130122, P.R. China, Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin 130062, P.R. China
Published online on: April 2, 2014 https://doi.org/10.3892/mmr.2014.2104
Pages: 2521-2527

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Brucellosis is a worldwide human and animal infectious disease, and the effective methods of its control are immunisation of animals by vaccination and elimination. Brucella abortus S19 is one of the popular vaccines with virulence in the control of cattle Brucellosis. In the present study, allelic exchange plasmids of wzm and wzt genes and partial knockout mutants of wzm and wzt were constructed to evaluate the resulting difference in virulence of B. abortus S19. PCR analysis revealed that the target genes were knocked out. The mutants were rough mutants and they could be differentiated from natural infection by the Rose Bengal plate and standard agglutination tests. The molecular weights of lipopolysaccharides of the Δwzm and Δwzt mutants were clustered between 25 and 40 kDa, and 30 and 35 kDa separately, and were markedly different from those in B. abortus S19. The virulence of B. abortus Δwzm and Δwzt was decreased compared with that of B. abortus S19 in mice. All these results identified that there were several differences between the wzm and wzt genes on lipopolysaccharide synthesis and on the virulence of B. abortus.

View Figures

View References

1	Cardoso PG, Macedo GC, Azevedo V and Oliveira SC: Brucella spp noncanonical LPS: structure, biosynthesis, and interaction with host immune system. Microb Cell Fact. 5:132006. View Article : Google Scholar : PubMed/NCBI
2	Rambow-Larsen AA, Petersen EM, Gourley CR and Splitter GA: Brucella regulators: self-control in hostile environment. Trends Microbiol. 17:371–377. 2009. View Article : Google Scholar : PubMed/NCBI
3	Pappas G, Papadimitriou P, Akritidis N, Christou L and Tsianos EV: The new global map of human brucellosis. Lancet Infect Dis. 6:91–99. 2006. View Article : Google Scholar : PubMed/NCBI
4	Ficht TA, Kahl-McDonagh MM, Arenas-Gamboa AM and Rice-Ficht AC: Brucellosis: the case for live, attenuated vaccines. Vaccine. 27:D40–D43. 2009. View Article : Google Scholar : PubMed/NCBI
5	Spink WW, Hall JW, Finstad J and Mallet E: Immunization with viable Brucella organisms. Results of a safety test in humans. Bull World Health Organ. 26:409–419. 1962.PubMed/NCBI
6	Fugier E, Pappas G and Gorvel JP: Virulence factors in brucellosis: implications for aetiopathogenesis and treatment. Expert Rev Mol Med. 9:1–10. 2007. View Article : Google Scholar : PubMed/NCBI
7	Bundle DR, Cherwonogrodzky JW, Gidney MA, Meikle PJ, Perry MB and Peters T: Definition of Brucella A and M epitopes by monoclonal typing reagents and synthetic oligosaccharides. Infect Immun. 57:2829–2836. 1987.PubMed/NCBI
8	Weynants V, Gilson D, Cloeckaert A, Tibor A, Denoel PA, Godfroid F, Limet JN and Letesson JJ: Characterization of smooth lipopolysaccharides and O polysaccharides of Brucella species by competition binding assays with monoclonal antibodies. Infect Immun. 65:1939–1943. 1997.PubMed/NCBI
9	Ugalde JE, Comerci DJ, Leguizamón MS and Ugalde RA: Evaluation of Brucella abortus phosphoglucomutase (pgm) mutant as a new live rough-phenotype vaccine. Infect Immun. 71:6264–6269. 2003.PubMed/NCBI
10	Lapaque N, Moriyon I, Moreno E and Gorvel JP: Brucella lipopolysaccharide acts as a virulence factor. Curr Opin Microbiol. 8:60–66. 2005. View Article : Google Scholar : PubMed/NCBI
11	Fernandez-Prada CM, Zelazowska EB, Nikolich M, Hadfield TL, Roop RM 2nd, Robertson GL and Hoover DL: Interactions between Brucella melitensis and human phagocytes: bacterial surface O-Polysaccharide inhibits phagocytosis, bacterial killing, and subsequent host cell apoptosis. Infect Immun. 71:2110–2119. 2003. View Article : Google Scholar
12	Jiménez de Bagüés MP, Terraza A, Gross A and Dornand J: Different responses of macrophages to smooth and rough Brucella spp: relationship to virulence. Infect Immun. 72:2429–2433. 2004.PubMed/NCBI
13	Moriyón I, Grilló MJ, Monreal D, González D, Marín C, López-Goñi I, Mainar-Jaime RC, Moreno E and Blasco JM: Rough vaccines in animals brucellosis: structural and genetic basis and present status. Vet Res. 35:1–38. 2004.PubMed/NCBI
14	Adone R, Ciuchini F, Marianelli C, Tarantino M, Pistoia C, Marcon G, Petrucci P, Francia M, Riccardi G and Pasquali P: Protective properties of rifampin-resistant rough mutants of Brucella melitensis. Infect Immun. 73:4198–4204. 2005. View Article : Google Scholar : PubMed/NCBI
15	Haag AF, Myka KK, Arnold MF, Caro-Hernández P and Ferguson GP: Importance of lipopolysaccharide and cyclic β-1,2-glucans in Brucella-mammalian infections. Int J Microbiol. 2010:1245092010.
16	Cloeckaert A, Grayon M, Verger JM, Letesson JJ and Godfroid F: Conservation of seven genes involved in the biosynthesis of the lipopolysaccharide O-side chain in Brucella spp. Res Microbiol. 151:209–216. 2000. View Article : Google Scholar : PubMed/NCBI
17	Godfroid F, Cloeckaert A, Taminiau B, Danese I, Tibor A, de Bolle X, Mertens P and Letesson JJ: Genetic organization of the lipopolysaccharide O-antigen biosynthesis region of Brucella melitensis 16M (wbk). Res Microbiol. 151:655–668. 2000. View Article : Google Scholar : PubMed/NCBI
18	Allen CA, Adams LG and Ficht TA: Transposon-derived Brucella abortus rough mutants are attenuated and exhibit reduced intracellular survival. Infect Immun. 66:1008–1016. 1998.PubMed/NCBI
19	Tsai CM and Frasch CE: A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 119:115–119. 1982. View Article : Google Scholar : PubMed/NCBI
20	Ried JL and Collmer A: An nptI-sacB-sacR cartridge for constructing directed, unmarked mutations in gram-negative bacteria by marker exchange-eviction mutagenesis. Gene. 57:239–246. 1987. View Article : Google Scholar : PubMed/NCBI
21	Orduna A, Almaraz A, Prado A, Gutierrez MP, Garcia-Pascual A, Dueñas A, Cuervo M, Abad R, Hernández B, Lorenzo B, Bratos MA and Torres AR: Evaluation of an immunocapture-agglutination test (Brucellacapt) for serodiagnosis of human brucellosis. J Clin Microbiol. 38:4000–4005. 2000.PubMed/NCBI
22	Seleem MN, Boyle SM and Sriranganathan N: Brucellosis: a re-emerging zoonosis. Vet Microbiol. 140:392–398. 2010. View Article : Google Scholar : PubMed/NCBI
23	Raetz CR and Whitfield C: Lipopolysaccharide endotoxins. Annu Rev Biochem. 71:635–700. 2002. View Article : Google Scholar : PubMed/NCBI
24	González D, Grilló MJ, De Miguel MJ, Ali T, Arce-Gorvel V, Delrue RM, Conde-Alvarez R, Muñoz P, López-Goñi I, Iriarte M, Marín CM, Weintraub A, Widmalm G, Zygmunt M, Letesson JJ, Gorvel JP, Blasco JM and Moriyón I: Brucellosis vaccines: assessment of Brucella melitensis lipopolysaccharide rough mutants defective in core and O-polysaccharide synthesis and export. PLoS One. 3:e27602008.
25	Langford MJ and Myers RC: Difficulties associated with the development and licensing of vaccines for protection against bio-warfare and bio-terrorism. Dev Biol (Basel). 110:107–112. 2002.PubMed/NCBI