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1.
BMC Vet Res ; 15(1): 304, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31438945

RESUMO

BACKGROUND: Brucellosis is a worldwide zoonotic infectious disease that is transmitted in various ways and causes great harm to humans and animals. The brucellosis pathogen is Brucella, which mainly resides in macrophage cells and survives and replicates in host cells. However, the mechanisms underlying Brucella survival in macrophage cells have not been thoroughly elucidated to date. Peroxiredoxin 6 (Prdx6) is a bifunctional protein that shows not only GSH peroxidase activity but also phospholipase A2 activity and plays important roles in combating oxidative damage and regulating apoptosis. RESULTS: Recombinant mouse (Mus musculus) Prdx6 (MmPrdx6) was expressed and purified, and monoclonal antibodies against MmPrdx6 were prepared. Using the Brucella suis S2 strain to infect RAW264.7 murine macrophages, the level of intracellular Prdx6 expression first decreased and later increased following infection. Overexpressing Prdx6 in macrophages resulted in an increase in B. suis S2 strain levels in RAW264.7 cells, while knocking down Prdx6 reduced the S2 levels in cells. CONCLUSIONS: Host Prdx6 can increase the intracellular survival of B. suis S2 strain and plays a role in Brucella infection.


Assuntos
Brucella suis/fisiologia , Brucelose/microbiologia , Peroxirredoxina VI/metabolismo , Animais , Feminino , Regulação da Expressão Gênica/fisiologia , Técnicas de Silenciamento de Genes , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Células RAW 264.7
2.
Microb Pathog ; 116: 180-188, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29331367

RESUMO

The interleukin-1 family is an important component of the innate immune system and plays an important role in regulating immune responses on the invasion of intracellular parasites in the acquired immune system. Interleukin 1ß (IL-1ß) is one of the members of the IL-1 family that predominantly activates downstream signaling pathways to play immunological functions of stimulating T and B lymphocyte activation and promoting the various syntheses of inflammatory substances in conjunction with other cytokines. Here, a full-length IL-1ß cDNA (OaIL-1ß) of sheep (Ovis aries) was cloned using rapid amplification of cDNA ends (RACE), which consists of 1494 bp and contains a 5'-UTR region with a length of 83 bp, a complete ORF of 801 bp in length, and a 3'-UTR region with a length of 642 bp. Recombinant protein OaIL-1ß was expressed and purified, and the monoclonal antibody against IL-1ß of sheep is prepared. Western blotting results showed that the sheep IL-1ß protein was detected in the heart, liver, lung, kidney, stomach, intestine, muscle, lymph nodes and leukocytes with the highest expression in the muscle and the lowest expression in the lung. Different bacteria treating sheep white blood cells induced differential expression of OaIL-1ß. Compared with the normal sheep, OaIL-1ß in the buffy coat was differentially expressed in the Brucella melitensis-challenged group and the B. suis S2 strain-inoculated group. However, whether IL-1ß may be considered as a molecular biomarker for differing Brucella-infected animals from brucellosis-vaccinated animals or not need to be further studied.


Assuntos
Brucelose/veterinária , Perfilação da Expressão Gênica , Interleucina-1beta/biossíntese , Interleucina-1beta/genética , Doenças dos Ovinos/patologia , Carneiro Doméstico , Estruturas Animais/patologia , Animais , Brucella melitensis/imunologia , Brucella suis/imunologia , Brucelose/patologia , Clonagem Molecular , Expressão Gênica , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Ovinos
3.
Vet Microbiol ; 186: 126-38, 2016 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-27016767

RESUMO

Acinetobacter baumannii is a Gram-negative bacillus that causes nosocomial infections, such as bacteremia, pneumonia, and meningitis and urinary tract and wound infections. In the present study, the surface antigen protein 1 (SurA1) gene of A. baumannii strain CCGGD201101 was identified, cloned and expressed, and then its roles in fitness and virulence were investigated. Virulence was observed in the human lung cancer cell lines A549 and HEp-2 at one week after treatment with recombinant SurA1. One isogenic SurA1 knock-out strain, GR0015, which was derived from the A. baumannii strain CCGGD201101 isolated from diseased chicks in a previous study, highlighted the effect of SurA1 on fitness and growth. Its growth rate in LB broth and killing activity in human sera were significantly decreased compared with strain CCGGD201101. In the Galleria mellonella insect model, the isogenic SurA1 knock-out strain exhibited a lower survival rate and decreased dissemination. These results suggest that SurA1 plays an important role in the fitness and virulence of A. baumannii.


Assuntos
Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/fisiologia , Acinetobacter baumannii/patogenicidade , Antígenos de Superfície/genética , Antígenos de Superfície/metabolismo , Virulência/genética , Acinetobacter baumannii/genética , Acinetobacter baumannii/crescimento & desenvolvimento , Sequência de Aminoácidos , Animais , Anticorpos Antibacterianos/imunologia , Sequência de Bases , Biofilmes , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Galinhas , Regulação da Expressão Gênica , Técnicas de Inativação de Genes , Humanos , Dados de Sequência Molecular , Mariposas/microbiologia , Plasmídeos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/toxicidade
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