The Brucella abortus phosphoglycerate kinase mutant is highly attenuated and induces protection superior to that of vaccine strain 19 in immunocompromised and immunocompetent mice.
Brucella abortus is a facultative intracellular bacterial pathogen that causes abortion in domestic animals and undulant fever in humans. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that can function as virulence factors to better understand the host-pathogen interplay. Herein, we identified the gene encoding the phosphoglycerate kinase (PGK) of B. abortus strain 2308. To test the role of PGK in Brucella pathogenesis, a pgk deletion mutant was constructed. Replacement of the wild-type pgk by recombination was demonstrated by Southern and Western blot analyses. The B. abortus Delta pgk mutant strain exhibited extreme attenuation in bone marrow-derived macrophages and in vivo in BALB/c, C57BL/6, 129/Sv, and interferon regulatory factor-1 knockout (IRF-1 KO) mice. Additionally, at 24 h postinfection the Delta pgk mutant was not found within the same endoplasmic reticulum-derived compartment as the wild-type bacteria, but, instead, over 60% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP1. Furthermore, the B. abortus Delta pgk deletion mutant was used as a live vaccine. Challenge experiments revealed that the Delta pgk mutant strain induced protective immunity in 129/Sv or IRF-1 KO mice that was superior to the protection conferred by commercial strain 19 or RB51. Finally, the results shown here demonstrated that Brucella PGK is critical for full bacterial virulence and that a Delta pgk mutant may serve as a potential vaccine candidate in future studies.
Trant
Cyntia G M C
CG
Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Pampulha, Belo Horizonte, MG, Brazil.
Lacerda
Thais L S
TL
Carvalho
Natalia B
NB
Azevedo
Vasco
V
Rosinha
Gracia M S
GM
Salcedo
Suzana P
SP
Gorvel
Jean-Pierre
JP
Oliveira
Sergio C
SC
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002004:Brucella Vaccine / Q000235:genetics / Q000276:immunology*; D002003:Brucella abortus / Q000201:enzymology* / Q000276:immunology / Q000472:pathogenicity*; D002006:Brucellosis / Q000276:immunology / Q000517:prevention & control*; D002478:Cells, Cultured; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010735:Phosphoglycerate Kinase / Q000172:deficiency*; D017422:Sequence Analysis, DNA; D013154:Spleen / Q000382:microbiology; D016019:Survival Analysis; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology
D001426:Bacterial Proteins; D002004:Brucella Vaccine; D004269:DNA, Bacterial; D014613:Vaccines, Attenuated; D010735:Phosphoglycerate Kinase