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1.
Vet Microbiol ; 159(1-2): 130-40, 2012 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-22483850

RESUMO

Brucella spp. are gram-negative intracellular bacterial pathogens that cause chronic infections. Brucella virulence factors include a type IV secretion system (T4SS) and its lipopolysaccharide (LPS), which are essential for persistence. However, the role of the virB-encoded T4SS has not been investigated in naturally rough Brucella species such as Brucella ovis. In this study, male 6-week old BALBc mice were infected with B. ovis, Brucella abortus, and their respective ΔvirB2 mutant strains. During early infection, B. ovis and B. abortus wild type strains were similarly recovered from spleen. Interestingly, in contrast to ΔvirB2 B. abortus that was recovered at similar levels when compared to the wild type strain, the ΔvirB2 B. ovis was markedly attenuated as early as 24h post infection (hpi). The ΔvirB2 B. ovis was unable to survive and multiply in murine peritoneal macrophages and extracellularly within the peritoneal cavity at 12 and 24 hpi with lower splenic colonization than the parental strain at 6, 12 and 24 hpi. In contrast, wild type B. abortus and ΔvirB2 B. abortus had a similar kinetics of infection in this model. As expected, the T4SS was essential for intracellular replication of smooth and rough strains in RAW macrophages at 48 hpi. These results suggest that T4SS is important for survival of B. ovis in murine model, and that a T4SS deficient B. ovis strain is cleared at earlier stages of infection when compared to a similar B. abortus mutant.


Assuntos
Sistemas de Secreção Bacterianos/fisiologia , Brucella ovis/genética , Brucella ovis/metabolismo , Brucelose/microbiologia , Animais , Sistemas de Secreção Bacterianos/genética , Brucella abortus/fisiologia , Brucella ovis/crescimento & desenvolvimento , Linhagem Celular , Lipopolissacarídeos/metabolismo , Macrófagos Peritoneais/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Viabilidade Microbiana , Baço/microbiologia , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
2.
Infect Immun ; 79(4): 1706-17, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21300772

RESUMO

Brucella ovis is a major cause of reproductive failure in sheep, which is associated with epididymitis and infertility in rams. Importantly, B. ovis is one of the few Brucella species that is not zoonotic. Due to the scarcity of studies on B. ovis infection, a murine model of infection was developed. The roles of B. ovis genes encoding a putative hemagglutinin and an ABC transporter were investigated in the mouse model. The kinetics of B. ovis infection were similar in BALB/c and C57BL/6 mice, and both strains of mice developed multifocal microgranulomas in the liver and spleen, but only minimal colonization and histopathological changes were observed in the genital tract. Therefore, the mouse was considered a suitable infection model for B. ovis but not for B. ovis-induced genital disease. Two mutant strains were generated in this study (the ΔabcAB and Δhmg strains). The B. ovis ΔabcAB strain was attenuated in the spleens and livers of BALB/c mice compared to the wild-type (WT) strain (P < 0.001). Conversely, the Δhmg strain infected mice at the same level as WT B. ovis, suggesting that a putative hemagglutinin is not required for B. ovis pathogenesis. Additionally, the ΔabcAB strain did not survive in peritoneal macrophages, extracellularly in the peritoneal cavity, or in RAW 264.7 macrophages. Moreover, infection with the ΔabcAB strain was not lethal for male regulatory factor 1-knockout mice, whereas infection with the B. ovis WT strain was 100% lethal within 14 days postinfection. These results confirm that the predicted ABC transporter is required for the full virulence and survival of B. ovis in vivo.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Brucella ovis/genética , Brucella ovis/patogenicidade , Fatores de Virulência/metabolismo , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brucelose/genética , Brucelose/metabolismo , Brucelose/patologia , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Virulência/genética
3.
Vet J ; 184(2): 146-55, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-19733101

RESUMO

Bovine brucellosis is one of the most important zoonotic diseases worldwide, and is of particular significance in developing countries. The disease, which results in serious economic losses due to late term abortion, stillborn and weakly calves, is caused by Gram negative coccobacilli bacteria of the genus Brucella. Lesions consist of necrotic placentitis and interstitial mastitis in pregnant cows, and fibrinous pleuritis with interstitial pneumonia in aborted fetuses and newborn calves. This article considers the pathogenesis of Brucella abortus and reviews the ability of the pathogen to invade phagocytic and non-phagocytic host cells, resist the acidified intraphagosomal environment, and inhibit phagosome-lysosome fusion. Significant aspects of innate and adaptive immunity against brucellosis are also discussed.


Assuntos
Aborto Animal/microbiologia , Brucella abortus/patogenicidade , Brucelose Bovina/microbiologia , Complicações Infecciosas na Gravidez/patologia , Animais , Brucelose Bovina/patologia , Bovinos , Feminino , Feto/microbiologia , Feto/patologia , Imuno-Histoquímica , Masculino , Gravidez
4.
Vet Immunol Immunopathol ; 127(3-4): 332-9, 2009 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-19084282

RESUMO

Nramp1 (Slc11a1) is linked to resistance to Leishmania in mice, but its role in canine leishmaniasis is not clear. In this study we sequenced the Nramp1 cDNA from dogs whose macrophages allowed or restricted intracellular growth of Leishmania chagasi. Peripheral blood monocyte-derived macrophages were isolated from 29 dogs, cultured and inoculated with L. chagasi. This approach resulted in the identification of dogs whose macrophages were resistant or susceptible to L. chagasi. Nramp1 cDNA sequences of these dogs were identical. mRNA levels of Nramp1, IFNgamma, IL-4 and the subunit p35 of IL-12 were assessed in the spleen of naturally infected symptomatic and asymptomatic dogs in comparison to uninfected controls. Although not statistically significant, asymptomatic dogs had a tendency for higher levels of Nramp1 mRNA (p = 0.11). Expression of Nramp1 was then compared between phenotypically resistant and susceptible dogs, without any significant difference between these groups.


Assuntos
Proteínas de Transporte de Cátions/genética , DNA Complementar/genética , Doenças do Cão/genética , Predisposição Genética para Doença , Leishmaniose Visceral/veterinária , Animais , Sequência de Bases , Proteínas de Transporte de Cátions/imunologia , Cães , Regulação da Expressão Gênica/imunologia , Leishmaniose Visceral/genética , Leishmaniose Visceral/imunologia , Macrófagos/parasitologia
5.
Infect Immun ; 76(5): 1897-907, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18316388

RESUMO

Brucellosis is still a widespread zoonotic disease. Very little is known about the interaction between Brucella abortus and trophoblastic cells, which is essential for better understanding the pathogenesis of the Brucella-induced placentitis and abortion, a key event for transmission of the disease. The goal of this study was to evaluate the profile of gene expression by bovine trophoblastic cells during infection with B. abortus. Explants of chorioallantoic membranes were inoculated with B. abortus strain 2308. Microarray analysis was performed at 4 h after infection, and expression of cytokines and chemokines by trophoblastic cells was assessed by real-time reverse transcription-PCR at 6 and 12 h after inoculation. In addition, cytokine and chemokine expression in placentomes from experimentally infected cows was evaluated. Expression of proinflammatory genes by trophoblastic cells was suppressed at 4 h after inoculation, whereas a significant upregulation of CXC chemokines, namely, CXCL6 (GCP-2) and CXCL8 (interleukin 8), was observed at 12 but not at 6 h after inoculation. Placentomes of experimentally infected cows had a similar profile of chemokine expression, with upregulation of CXCL6 and CXCL8. Our data indicate that B. abortus modulates the innate immune response by trophoblastic cells, suppressing the expression of proinflammatory mediators during the early stages of infection that is followed by a delayed and mild expression of proinflammatory chemokines, which is similar to the profile of chemokine expression in the placentomes of experimentally infected cows. This trophoblastic response is likely to contribute to the pathogenesis of B. abortus-induced placentitis.


Assuntos
Brucella abortus/imunologia , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/imunologia , Trofoblastos/imunologia , Trofoblastos/microbiologia , Animais , Bovinos , Citocinas/biossíntese , Citocinas/genética , Regulação para Baixo , Feminino , Análise de Sequência com Séries de Oligonucleotídeos , Placenta/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Trofoblastos/metabolismo , Regulação para Cima
6.
Infect Immun ; 75(5): 2493-9, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17353292

RESUMO

The NRAMP1 gene encodes a divalent cation transporter, located in the phagolysosomal membrane of macrophages, that has been associated with resistance to intracellular pathogens. In cattle, natural resistance against brucellosis has been associated with polymorphisms at the 3' untranslated region (3'UTR) of the NRAMP1 gene, which are detectable by single-strand conformational analysis (SSCA). This study aimed to evaluate the association between NRAMP1 3'UTR polymorphisms and resistance against bovine brucellosis in experimental and natural infections. In experimentally infected pregnant cows, abortion occurred in 42.1% of cows with a resistant genotype (SSCA(r); n = 19) and in 43.1% of those with a susceptible genotype (SSCA(s); n = 23). Furthermore, no association between intensity of pathological changes and genotype was detected. In a farm with a very high prevalence of bovine brucellosis, the percentages of strains of the SSCA(r) genotype were 86 and 84% in serologically positive (n = 64) and negative (n = 36) cows, respectively. Therefore, no association was found between the NRAMP1-resistant allele and the resistant phenotype in either experimental or naturally occurring brucellosis. To further support these results, bacterial intracellular survival was assessed in bovine monocyte-derived macrophages from cattle with either the resistant or susceptible genotype. In agreement with our previous results, no difference was observed in the rates of intracellular survival of B. abortus within macrophages from cattle with susceptible or resistant genotypes. Taken together, these results indicate that these polymorphisms at the NRAMP1 3'UTR do not affect resistance against B. abortus in cattle and that they are therefore not suitable markers of natural resistance against bovine brucellosis.


Assuntos
Regiões 3' não Traduzidas/genética , Brucella abortus/classificação , Brucella abortus/patogenicidade , Brucelose Bovina/imunologia , Proteínas de Transporte de Cátions/genética , Polimorfismo Genético , Aborto Animal/genética , Aborto Animal/imunologia , Aborto Animal/microbiologia , Animais , Brucella abortus/genética , Brucelose Bovina/genética , Brucelose Bovina/microbiologia , Bovinos , Células Cultivadas , Feminino , Predisposição Genética para Doença , Genótipo , Macrófagos/microbiologia , Masculino , Doenças Placentárias/genética , Doenças Placentárias/imunologia , Doenças Placentárias/microbiologia , Doenças Placentárias/veterinária , Polimorfismo Conformacional de Fita Simples , Gravidez
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