RESUMO
Trueperella pyogenes (T. pyogenes) is a versatile and ingenious bacterium that causes severe suppurative injuries in lots of economically important ruminants. The underlying pathogenesis of T. pyogenes infection remains poorly understood. In the current study, we performed transcriptome sequencing of mouse blood tissue infected with T. pyogenes. A total of 36.73 G clean data were collected, and 136 differentially expressed genes were obtained in the infection group compared to the control group. In addition, we found that the E3 ubiquitin ligase Cblb exhibited significant upregulation in the infection groups compared to the control group. Mechanistically, T. pyogenes infection markedly enhanced the expression of Cblb and regulated the host defense response. Inhibiting Cblb expression with Cblb siRNA impaired the inflammatory response and reduced the effect of phagocytosis in RAW264.7 murine macrophages. Intriguingly, overexpression of Cblb induced a strong inflammatory response and enhanced phagocytosis against T. pyogenes infection in macrophages. More importantly, the overexpression of Cblb significantly reduced the bacterial load and protected mice from the T. pyogenes infections. Therefore, our findings reveal that Cblb is a novel and potential regulator in response to T. pyogenes infection and shed new light on the development of promising treatments against T. pyogenes-related diseases.
Assuntos
Actinomycetaceae , Infecções por Actinomycetales , Proteínas Proto-Oncogênicas c-cbl , Ubiquitina-Proteína Ligases , Animais , Camundongos , Proteínas Adaptadoras de Transdução de Sinal/genética , Macrófagos , Fagocitose/genética , Proteínas Proto-Oncogênicas c-cbl/genética , Transcriptoma , Ubiquitina-Proteína Ligases/genética , Actinomycetaceae/fisiologia , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologiaRESUMO
Bacterial vaginosis (BV) is a common type of vaginal inflammation caused by a proliferation of pathogenic bacteria, among which Mobiluncus curtisii. In our previous studies on M. curtisii genome, we identified the presence of a genomic fragment encoding a 25 kDa pore-forming toxin, the CAMP factor, which is known to be involved in the synergistic lysis of erythrocytes namely CAMP reaction. However, whether this hypothetical gene product has hemolytic activity is unknown. Moreover, its relative structure and function are not yet solved. Here we found that the M. curtisii CAMP factor is a monomer at pH 4.4 and oligomer at pH > 4.6. Hemolysis assays showed that M. curtisii CAMP factor could lyse sheep red blood cells efficiently in pH 5.4-7.4. Negative staining electron microscope analysis of the CAMP factor revealed ring-like structures at pH above 4.6. Additionally, the crystal structure of M. curtisii CAMP factor, determineded at 1.85 Å resolution, reveals a 5 + 3 helix motif. Further functional analysis suggested that the structural rearrangement of the N-terminal domain might be required for protein function. In conclusion, this structure-function relationship study of CAMP factor provides a new perspective of the M. curtisii role in BV development.
Assuntos
Proteínas de Bactérias/química , Mobiluncus/química , Simulação de Dinâmica Molecular , Proteínas Citotóxicas Formadoras de Poros/química , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/metabolismo , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Eritrócitos/metabolismo , Eritrócitos/microbiologia , Feminino , Humanos , Mobiluncus/genética , Mobiluncus/metabolismo , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Domínios Proteicos , Ovinos , Relação Estrutura-Atividade , Vaginose Bacteriana/genética , Vaginose Bacteriana/metabolismoRESUMO
Rhodococcus equi pneumonia is a major cause of morbidity and mortality in neonatal foals. Much effort has been made to identify preventative measures and new treatments for R. equi with limited success. With a growing focus in the medical community on understanding the genetic basis of disease susceptibility, investigators have begun to evaluate the interaction of the genetics of the foal with R. equi. This review describes past efforts to understand the genetic basis underlying R. equi susceptibility and tolerance. It also highlights the genetic technology available to study horses and describes the use of this technology in investigating R. equi. This review provides readers with a foundational understanding of candidate gene approaches, single nucleotide polymorphism-based, and copy number variant-based genome-wide association studies, and next generation sequencing (both DNA and RNA).
Assuntos
Infecções por Actinomycetales/veterinária , Predisposição Genética para Doença , Doenças dos Cavalos/microbiologia , Rhodococcus equi , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/microbiologia , Animais , Doenças dos Cavalos/genética , CavalosRESUMO
The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme-g (CC-Lys-g) produced in Escherichia coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme-g plasmid DNA could be used as an immunostimulant to protect channel catfish against Aeromonas hydrophila infection. Recombinant CC-Lys-g produced in E. coli expression system exhibited significant (P < 0.05) lytic activity against Gram-positive Micrococcus lysodeikticus and Gram-negative A. hydrophila. When pcDNA3.2-vectored recombinant channel catfish lysozyme-g (pcDNA-Lys-g) was transfected in channel catfish gill cells G1B, the over-expression of pcDNA-Lys-g offered significant (P < 0.05) protection to G1B cells against A. hydrophila infection. When channel catfish were intraperitoneally injected with pcDNA-Lys-g along with an adjuvant QCDCR, the transcriptional level of Lys-g was significantly (P < 0.05) increased. When pcDNA-Lys-g injected fish was challenged with a highly virulent A. hydrophila strain AL-09-71, pcDNA-Lys-g offered 100% protection to channel catfish at two days post DNA injection. Macrophages of fish injected with pcDNA-Lys-g produced significantly (P < 0.05) higher amounts of reactive oxygen species and nitric oxide than that of fish injected with pcDNA vector alone at two days post DNA injection. Taken together, our results suggest that pcDNA-Lys-g could be used as a novel immunostimulant to offer immediate protection to channel catfish against A. hydrophila infection.
Assuntos
Infecções por Actinomycetales/veterinária , Doenças dos Peixes/prevenção & controle , Proteínas de Peixes/genética , Infecções por Bactérias Gram-Negativas/veterinária , Ictaluridae/genética , Ictaluridae/imunologia , Muramidase/genética , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/prevenção & controle , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/genética , Adjuvantes Imunológicos/metabolismo , Aeromonas hydrophila/imunologia , Aeromonas hydrophila/patogenicidade , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Escherichia coli/genética , Doenças dos Peixes/genética , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/prevenção & controle , Ictaluridae/metabolismo , Micrococcus/imunologia , Micrococcus/fisiologia , Dados de Sequência Molecular , Muramidase/química , Muramidase/metabolismo , Filogenia , Plasmídeos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência/veterináriaRESUMO
BACKGROUND: Bacterial vaginosis (BV) is the most common vaginal disorder of reproductive-age women. Yet the cause of BV has not been established. To uncover key determinants of BV, we employed a multi-omic, systems-biology approach, including both deep 16S rRNA gene-based sequencing and metabolomics of lavage samples from 36 women. These women varied demographically, behaviorally, and in terms of health status and symptoms. PRINCIPAL FINDINGS: 16S rRNA gene-based community composition profiles reflected Nugent scores, but not Amsel criteria. In contrast, metabolomic profiles were markedly more concordant with Amsel criteria. Metabolomic profiles revealed two distinct symptomatic BV types (SBVI and SBVII) with similar characteristics that indicated disruption of epithelial integrity, but each type was correlated to the presence of different microbial taxa and metabolites, as well as to different host behaviors. The characteristic odor associated with BV was linked to increases in putrescine and cadaverine, which were both linked to Dialister spp. Additional correlations were seen with the presence of discharge, 2-methyl-2-hydroxybutanoic acid, and Mobiluncus spp., and with pain, diethylene glycol and Gardnerella spp. CONCLUSIONS: The results not only provide useful diagnostic biomarkers, but also may ultimately provide much needed insight into the determinants of BV.
Assuntos
Infecções por Actinomycetales/diagnóstico , Infecções por Bactérias Gram-Positivas/diagnóstico , Metabolômica , RNA Ribossômico 16S/genética , Vagina/microbiologia , Doenças Vaginais/diagnóstico , Vaginose Bacteriana/diagnóstico , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/microbiologia , Adulto , DNA Bacteriano/genética , Feminino , Redes Reguladoras de Genes , Infecções por Bactérias Gram-Positivas/genética , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Hidroxibutiratos/metabolismo , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Pessoa de Meia-Idade , Mobiluncus/genética , Mobiluncus/isolamento & purificação , Reação em Cadeia da Polimerase , Doenças Vaginais/etiologia , Doenças Vaginais/metabolismo , Vaginose Bacteriana/complicações , Vaginose Bacteriana/microbiologia , Adulto JovemRESUMO
Rhodococcus equi is a facultative intracellular bacterial pathogen of horses; infected foals develop pyogranulomatous pneumonia, however adult horses are largely unaffected. R. equi infects and proliferates within host macrophages and dendritic cells (DCs). DCs initiate the appropriate adaptive immune response, thereby playing a critical role in determining the outcome of infection. Our aim was to identify genes that are differentially expressed in R. equi infected monocyte-derived DCs (mdDCs). Peripheral blood monocytes from mares and foals were used to derive mdDCs by culturing with recombinant equine IL-4 and recombinant human GM-CSF. RNA harvested 24h after infection with R. equi (ATCC 33701+) was used to perform suppression subtractive hybridization (SSH) experiments. Approximately 38 unique sequences were obtained from these experiments. Differential expression of 19 immunologically relevant genes was validated by PCR. These genes are characterized by the following functions: cell adhesion, chemotaxis/migration, immune/inflammatory response, ion transport, signal transduction, T-cell regulation, and vesicular transport. In summary, we identified several novel genes that are differentially expressed in foal and adult mdDCs in response to R. equi infection. These genes provide promising targets for further research into the host response to R. equi, and the susceptibility of foals to this disease.
Assuntos
Infecções por Actinomycetales/veterinária , Células Dendríticas/imunologia , Doenças dos Cavalos/genética , Doenças dos Cavalos/imunologia , Cavalos/genética , Cavalos/imunologia , Rhodococcus equi/imunologia , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Animais , Sequência de Bases , Primers do DNA/genética , Feminino , Expressão Gênica , Doenças dos Cavalos/microbiologia , Cavalos/microbiologia , Masculino , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Rhodococcus equi/patogenicidadeRESUMO
The globular C1q-domain-containing (C1qDC) proteins are a family of versatile pattern recognition receptors via their globular C1q (gC1q) domain to bind various ligands including several PAMPs on pathogens. In this study, a new gC1q-domain-containing protein (AiC1qDC-1) gene was cloned from Argopecten irradians by rapid amplification of cDNA ends (RACE) approaches and expressed sequence tag (EST) analysis. The full-length cDNA of AiC1qDC-1 was composed of 733bp, encoding a signal peptide of 19 residues and a typical gC1q domain of 137 residues containing all eight invariant amino acids in human C1qDC proteins and seven aromatic residues essential for effective packing of the hydrophobic core of AiC1qDC-1. The gC1q domain of AiC1qDC-1, which possessed the typical 10-stranded beta-sandwich fold with a jelly-roll topology common to all C1q family members, showed high homology not only to those of C1qDC proteins in mollusk but also to those of C1qDC proteins in human. The AiC1qDC-1 transcripts were mainly detected in the tissue of hepatopancreas and also marginally detectable in adductor, heart, mantle, gill and hemocytes by fluorescent quantitative real-time PCR. In the microbial challenge experiment, there was a significant up-regulation in the relative expression level of AiC1qDC-1 in hepatopancreas and hemocytes of the scallops challenged by fungi Pichia pastoris GS115, Gram-positive bacteria Micrococcus luteus and Gram-negative bacteria Listonella anguillarum. The recombinant AiC1qDC-1 (rAiC1qDC-1) protein displayed no obvious agglutination against M. luteus and L. anguillarum, but it aggregated P. pastoris remarkably. This agglutination could be inhibited by d-mannose and PGN but not by LPS, glucan or d-galactose. These results indicated that AiC1qDC-1 functioned as a pattern recognition receptor in the immune defense of scallops against pathogens and provided clues for illuminating the evolution of the complement classical pathway.
Assuntos
Infecções por Actinomycetales/imunologia , Complemento C1q/metabolismo , Infecções por Bactérias Gram-Negativas/imunologia , Hepatopâncreas/metabolismo , Listonella/imunologia , Micrococcus luteus/imunologia , Micoses/imunologia , Pectinidae/genética , Pichia/imunologia , Receptores de Reconhecimento de Padrão/metabolismo , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/metabolismo , Aglutinação , Sequência de Aminoácidos , Animais , Complemento C1q/química , Complemento C1q/genética , Complemento C1q/imunologia , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/metabolismo , Hemócitos/imunologia , Hemócitos/metabolismo , Hemócitos/microbiologia , Hepatopâncreas/imunologia , Hepatopâncreas/microbiologia , Humanos , Imunidade Inata , Listonella/patogenicidade , Micrococcus luteus/patogenicidade , Dados de Sequência Molecular , Micoses/genética , Micoses/metabolismo , Pectinidae/microbiologia , Filogenia , Pichia/patogenicidade , Dobramento de Proteína , Estrutura Terciária de Proteína/genética , Receptores de Reconhecimento de Padrão/química , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologiaRESUMO
Mendelian susceptibility to mycobacterial disease (MSMD) is a rare primary immunodeficiency associated with clinical disease caused by weakly virulent mycobacterial species. Interferon gamma receptor 1 (IFN-gammaR1) deficiency is a genetic etiology of MSMD. We describe the clinical and genetic features of a 7-year-old Italian boy suffering from MSMD associated with a complex phenotype, including neonatal hyperglycemia, neuromuscular disease, and dysmorphic features. The child also developed necrotizing pneumonia caused by Rhodococcus equi. The child is homozygous for a nonsense mutation in exon 3 of IFNGR1 as a result of paternal uniparental disomy (UPD) of the entire chromosome 6. This is the first reported case of uniparental disomy resulting in a complex phenotype including MSMD.
Assuntos
Anormalidades Múltiplas/genética , Cromossomos Humanos Par 6/genética , Infecções por Mycobacterium/genética , Receptores de Interferon/deficiência , Receptores de Interferon/genética , Dissomia Uniparental/genética , Infecções por Actinomycetales/genética , Criança , Códon sem Sentido , Análise Mutacional de DNA , Éxons , Pai , Feminino , Homozigoto , Humanos , Masculino , Linhagem , Fenótipo , Rhodococcus equi , Síndrome , Receptor de Interferon gamaRESUMO
In previous work, we found significant associations of horse polymorphic microsatellite and immunity-related (IR) gene markers with Rhodococcus equi infection of foals. Here, a statistically significant association between a single nucleotide polymorphism (SNP) within the interleukin 7 receptor-encoding gene (IL7R) with high R. equi burden in transtracheal aspirates was found (Fisher's F = 0.043, odds ratio: 8.00, 95% confidence interval: 1.127-56.795). Further positional and/or functional candidate genes investigated TLR2, IL13, IL17A, IL28R, TACE/ADAM 17 and GBP1, were not associated with infection in this study. SNPs analysed were found by sequencing and appropriate restriction fragment length polymorphism markers were developed. Their associations with R. equi infection were tested by genotyping thoroughbred foals from the original study. The association was confirmed by analysing genotypes composed with genes previously reported to be associated with R. equi infection in the same group.
Assuntos
Infecções por Actinomycetales/veterinária , Doenças dos Cavalos/imunologia , Imunidade/genética , Polimorfismo de Nucleotídeo Único , Receptores de Interleucina-7/genética , Rhodococcus equi/imunologia , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/microbiologia , Animais , Frequência do Gene , Predisposição Genética para Doença/genética , Genótipo , Doenças dos Cavalos/genética , Doenças dos Cavalos/microbiologia , Cavalos , Polimorfismo de Fragmento de Restrição , Rhodococcus equi/isolamento & purificação , Análise de Sequência de DNA , Traqueia/microbiologiaRESUMO
The objective of this pilot study was to compare the cytokine profile as well as cell-mediated and antibody responses of foals infected with a low inoculum of virulent Rhodococcus equi resulting in subclinical pneumonia to that of foals infected with a high inoculum resulting in severe clinical pneumonia. The mean (+/-SD) ratio of post-infection to pre-infection anti-R. equi IgG(T) concentration was significantly (P=0.002) higher in foals infected with the high inoculum (195+/-145; range 62-328) compared to foals infected with the low inoculum (3.9+/-4.5; range 0.5-11). Similarly, mean (+/-SD) ratio of post-infection to pre-infection IgM concentration was significantly (P=0.002) higher in foals infected with the high inoculum (12+/-4.0; range 7.4-14) compared to foals infected with the low inoculum (2.5+/-1.5; range 1.2-4.7). Proliferative responses to R. equi antigens as well as expression of mRNA for IL-2, IL-4, IL-10, and IFN-gamma in BLN were not significantly different between the two groups. There was a tendency (P=0.073) towards a higher IFN-gamma/IL-4 ratio in the low inoculum group. This study demonstrates that the size of inoculum modulates the IgG subisotype response and possibly the cytokine profile of foals.
Assuntos
Infecções por Actinomycetales/veterinária , Doenças dos Cavalos/imunologia , Rhodococcus equi , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias , Brônquios/imunologia , Contagem de Colônia Microbiana , Citocinas/genética , Doenças dos Cavalos/genética , Cavalos , Imunidade Celular , Imunidade Humoral , Imunoglobulina G/sangue , Imunoglobulina G/classificação , Imunoglobulina M/sangue , Linfonodos/imunologia , Ativação Linfocitária , Projetos Piloto , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Rhodococcus equi/imunologia , Rhodococcus equi/patogenicidadeRESUMO
An effective host response to Renibacterium salmoninarum, the etiologic agent of bacterial kidney disease, is poorly characterized. Using suppression subtractive hybridization, we exploited the difference in early host response in the pronephros of fish challenged by an attenuated strain (MT239) or a virulent strain (ATCC 33209) of R. salmoninarum. Among the 132 expressed sequence tag (EST) clones that were sequenced, 20 were selected for expression analysis at 24 and 72h after challenge. ESTs matching two interferon inducible genes (IFN-inducible GBP and VLIG1), the ligand GAS6, and the kinase VRK2 were upregulated in fish exposed to MT239, but downregulated or unchanged in fish exposed to 33209. A second group of ESTs matching genes involved in apoptosis (caspase 8) and immune function (IkappaBalpha, p47(phoX), EMR/CD97) were more slowly upregulated in fish exposed to 33209 compared to fish exposed to MT239. The ESTs displaying elevated expression in MT239-exposed fish may represent important cellular processes to bacterial challenge, and may be useful indicators of an effective host response to R. salmoninarum infection.
Assuntos
Micrococcaceae/genética , Micrococcaceae/imunologia , Salmão/genética , Salmão/imunologia , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/microbiologia , Animais , DNA Complementar/genética , Nefropatias/genética , Nefropatias/microbiologia , Fosfoproteínas/genética , Proteínas Ribossômicas/genética , Transcrição Gênica/genéticaRESUMO
Rhodococcus equi is an intracellular pathogen of macrophages that causes rhodococcal pneumonia in foals and immunocompromised people. Evidence exists that neutrophils play a vital role in resistance to infection with R. equi; however, the means by which neutrophils exert their effects have not been clearly defined. In addition to directly killing bacteria, neutrophils also may exert a protective effect by linking innate and adaptive immune responses. In the present study we evaluated the cytokine expression profiles of adult equine neutrophils in response to stimulation with isogenic strains of virulent and avirulent R. equi in vitro. After 2 and 4h incubation with virulent or avirulent R. equi, adult equine neutrophils expressed significantly (P<0.05) greater tumor necrosis factor alpha (TNFalpha), interleukin (IL)-12p40, IL-6, IL-8 and IL-23p19 mRNA, but not interferon gamma (IFNgamma) or IL-12p35 mRNA than unstimulated neutrophils. Furthermore, virulent R. equi induced significantly greater IL-23p19 mRNA than avirulent R. equi. These results demonstrate that R. equi-stimulated neutrophils are a source of many proinflammatory cytokines. Furthermore, these results suggest that IL-23 may be preferentially expressed over IL-12 in response to exposure with R. equi, and that this response may be more strongly induced by virulent R. equi than avirulent R. equi. Collectively, the data presented herein suggest a non-phagocytic role for neutrophils that may influence the type of adaptive immune response to R. equi.
Assuntos
Citocinas/genética , Cavalos/genética , Cavalos/imunologia , Neutrófilos/imunologia , Rhodococcus equi/imunologia , Rhodococcus equi/patogenicidade , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Infecções por Actinomycetales/veterinária , Animais , Sequência de Bases , Primers do DNA/genética , Expressão Gênica , Perfilação da Expressão Gênica , Doenças dos Cavalos/genética , Doenças dos Cavalos/imunologia , Técnicas In Vitro , Subunidade p40 da Interleucina-12/genética , Subunidade p19 da Interleucina-23/genética , Interleucina-6/genética , Interleucina-8/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/genética , Virulência/imunologiaRESUMO
Renitelo breed is a cattle breed created at Kianjasoa station (Madagascar) by a triple crossing Malagasy Zebu x Limousine x Afrikander. This breed besides many valuable advantages, such as rapid growth and drought power, presents a huge disadvantage which is sensitivity to skin disease, dermatophilosis, previously known as streptotrichosis. This disease caused by Dermatophilus congolensis is one of the major threats for the population of Renitelo cattle. An allele of MHC gene has been shown to be dramatically associated to hypersensitivity to the disease in other cattle breed. To bring further information to tick borne disease clinical survey, mainly dermatophilosis, we wanted to verify if such allele could be found in this breed. Renitelo cattle included in this study were chosen for the presence of dermatophilosis lesions in more or less severe form (N = 17). These animals were blood sampled and a genetic analysis on the MHC gene BoLA-DRB3 was performed, by PCR amplification using BOD 31 & BOD 32 primers. Amplified products were analyzed by RFLP using enzymes. Restriction band profiles were characterized according to previously defined patterns. Three cows out of the 17 cattle analyzed for MHC gene presented the hypersensitive allele FDA. Two out of the three hypersensitive cows were pure breed while one was half breed. All the cows presented dermatophilosis lesions at least during rainy season but one of them particularly suffered from severe lesions covering all its body and died of the illness. This study shows that hypersensitivity allele found in other bovine breeds can be found in Renitelo breed. This result seemed to suggest that this characterization could be utilized in breeding program for this breed.
Assuntos
Infecções por Actinomycetales/veterinária , Doenças dos Bovinos/imunologia , Complexo Principal de Histocompatibilidade/genética , Polimorfismo de Fragmento de Restrição , Dermatopatias Bacterianas/veterinária , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Infecções por Actinomycetales/microbiologia , Alelos , Animais , Cruzamento , Bovinos , Doenças dos Bovinos/genética , Doenças dos Bovinos/microbiologia , Cruzamentos Genéticos , Suscetibilidade a Doenças/veterinária , Amplificação de Genes , Imunidade Inata , Madagáscar , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Dermatopatias Bacterianas/genética , Dermatopatias Bacterianas/imunologia , Dermatopatias Bacterianas/microbiologia , Infestações por Carrapato/veterináriaRESUMO
Rhodococcus equi is an important intracellular pathogen of horses, most commonly causing chronic, suppurative bronchopneumonia in foals. Although most foals likely are exposed to environmental R. equi within the 1st few days of life, only some develop R. equi pneumonia, and the basis of differences in susceptibility among foals currently is unknown. In this study, we investigated solute carrier family 11 member 1 (SLC11A1) gene sequences in the 5' untranslated region, exon 1, and a portion of intron 1 for variations in 3 equid species (horse, donkey, zebra) and compared variants within 3 independent horse breeding farms for associations with R. equi pneumonia by use of an age-matched case-control design. Seven novel variants in the 5'untranslated region were identified as specific for one or both of the non-horse equid species sampled. In addition, a single novel horse variant in the 5'untranslated region, -57C/T, was identified in 4 breeds. The -57C/T variant was found on 2 of the 3 farms with endemic R. equi pneumonia, representing 2 different horse breeds. Significant allelic and genotypic associations with susceptibility to R. equi pneumonia were observed for the -57C/T variant in foals from these farms. Although the functional impact of this novel variant remains to be determined, this study represents an important step in our understanding of natural resistance to R. equi foal pneumonia and other intracellular bacterial diseases affecting equids.
Assuntos
Infecções por Actinomycetales/veterinária , Proteínas de Transporte de Cátions/genética , Predisposição Genética para Doença/genética , Doenças dos Cavalos/genética , Doenças dos Cavalos/microbiologia , Pneumonia Bacteriana/veterinária , Rhodococcus equi , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/microbiologia , Animais , Variação Genética , Cavalos , Pneumonia Bacteriana/genética , Pneumonia Bacteriana/microbiologiaRESUMO
After i.v. inoculation with Rhodococcus aurantiacus, wild-type (WT) mice develop nonnecrotic, epithelioid granulomas. Because a high level of TNF-alpha is observed during the initial phase postinfection, we examined the extent to which TNF-alpha contributes to granulomatous inflammation using TNF-alpha gene-deficient (TNF-alpha(-/-)) mice. Despite a lack of R. aurantiacus proliferation, TNF-alpha(-/-) mice displayed high mortality rates within 5 days postinfection, as well as a high level of IL-6 in their spleens. Histological examination showed an absence of granuloma formation in TNF-alpha(-/-) mice. Pretreatment of TNF-alpha(-/-) mice with rTNF-alpha failed to restore this granuloma formation but accelerated bacterial removal and cellular recruitment. This rTNF-alpha administration also attenuated IL-6 production, resulting in increased survival rates of TNF-alpha(-/-) mice. Heat-killed R. aurantiacus induced in vitro enhanced mRNA expression and production of IL-6 in macrophages and DCs from TNF-alpha(-/-) mice when compared with WT controls, and treatment of TNF-alpha(-/-) mouse cells with rTNF-alpha decreased the IL-6 secretion. Moreover, anti-TNF-alpha or anti-IL-6 treatment increased IL-6 or TNF-alpha production by WT mouse cells, respectively. These data suggest that the production of TNF-alpha and IL-6 can be negatively regulated by each other. Administration of rIFN-gamma to TNF-alpha(-/-) mice caused immature granulomas in livers, and treatment with both rTNF-alpha and rIFN-gamma led to the formation of mature granulomas. Overall, TNF-alpha appears crucial for bacterial clearance, cellular recruitment, and granuloma formation. The balance between TNF-alpha and IL-6 during the early phase of infection controls the development of the inflammatory response to R. aurantiacus infection.
Assuntos
Infecções por Actinomycetales/imunologia , Granuloma/imunologia , Granuloma/microbiologia , Interleucina-6/fisiologia , Rhodococcus/imunologia , Fator de Necrose Tumoral alfa/fisiologia , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/mortalidade , Animais , Células Cultivadas , Citocinas/biossíntese , Citocinas/genética , Citocinas/metabolismo , Regulação para Baixo/genética , Regulação para Baixo/imunologia , Feminino , Granuloma/genética , Granuloma/mortalidade , Inflamação/genética , Inflamação/imunologia , Inflamação/microbiologia , Inflamação/mortalidade , Interleucina-6/antagonistas & inibidores , Interleucina-6/biossíntese , Cinética , Fígado/imunologia , Fígado/metabolismo , Fígado/microbiologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Mensageiro/biossíntese , Rhodococcus/crescimento & desenvolvimento , Baço/imunologia , Baço/metabolismo , Baço/microbiologia , Baço/patologia , Fator de Necrose Tumoral alfa/deficiência , Fator de Necrose Tumoral alfa/genéticaRESUMO
A quantitative polymerase chain reaction (QPCR) assay has been developed to detect varying levels of Renibacterium salmoninarum, the causative agent of bacterial kidney disease. This assay allows for the direct enumeration of bacterial DNA or RNA copy number within tissues and body fluids. The assay can be applied non-lethally and can be used to determine whether R. salmoninarum is transcriptionally active. The presence of R. salmoninarum in kidney tissues from 430 chinook salmon collected from five Idaho Fish and Game operated hatcheries was initially evaluated using the widely employed enzyme-linked immunosorbent assay (ELISA) with two sets of Kirkegaard and Perry Laboratories polyclonal antibodies, 'mother batches' 1 and 2. The same tissue samples were then analysed using the novel QPCR assay and the results compared. At moderate to high levels of infection [optical density (OD > 0.5)], ELISA values and estimated DNA copy number were highly correlated (r(2) > 0.80), although correlation to specific antibody batches varied. However, lower ELISA values (OD < 0.5) observed with either antibody batch did not correlate well with the QPCR assay (R(2) Assuntos
Infecções por Actinomycetales/veterinária
, Aquicultura/métodos
, Doenças dos Peixes/genética
, Doenças dos Peixes/microbiologia
, Micrococcaceae/genética
, Reação em Cadeia da Polimerase/veterinária
, Salmão
, Infecções por Actinomycetales/genética
, Animais
, Primers do DNA
, Ensaio de Imunoadsorção Enzimática
, Rim/microbiologia
, Reação em Cadeia da Polimerase/métodos
RESUMO
Rhodococcus equi is an intracellular bacterium that causes pneumonia in immunocompromised people and foals. The Nramp1 gene influences susceptibility to a variety of intracellular bacteria (including mycobacterial species), but not to Mycobacterium tuberculosis. In this study, we demonstrate that mice functionally deleted of the Nramp1 gene were not more susceptible to infection with virulent R. equi (ATCC 33701) than wild-type mice. Susceptibility of mice to infection with the intracellular bacterium R. equi is more similar to that of M. tuberculosis than to other intracellular bacteria, including other mycobacteria.
Assuntos
Proteínas de Transporte de Cátions/genética , Rhodococcus equi/patogenicidade , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Infecções por Actinomycetales/microbiologia , Animais , Proteínas de Transporte de Cátions/deficiência , Proteínas de Transporte de Cátions/fisiologia , Contagem de Colônia Microbiana , Deleção de Genes , Cavalos , Humanos , Camundongos , Camundongos Knockout , Rhodococcus equi/isolamento & purificação , VirulênciaRESUMO
White piedra is a fungal infection of the hair shaft caused by Trichosporon beigelii. A synergistic coryneform bacterial infection is often present with T beigelii. White piedra, although not commonly reported to infect scalp hair in North America, is an important consideration in the differential diagnosis of scalp hair concretions. We report a case of white piedra of scalp hair with synergistic coryneform bacterial infection in two sisters, both US natives. Culture and light and electronmicroscopic evidence of the synergistic infection are presented.
Assuntos
Infecções por Actinomycetales/epidemiologia , Piedra/epidemiologia , Infecções por Actinomycetales/genética , Adolescente , Humanos , Piedra/genéticaRESUMO
Bovine dermatophilosis is a severe skin infection of tropical ruminants inducing a severe loss in productivity and a 15% mortality rate. This disease is caused by the actinomycete bacterium Dermatophilus congolensis associated with the tick Amblyomma variegatum. Currently there are no prospects for a vaccine, and acaricide or antibiotic control is hampered by the development of chemoresistance. Animal breeders have observed that dermatophilosis susceptibility seems to be determined genetically, and we previously identified a BoLA-DRB3-DQB class II haplotype marker for high (R2= 0.96) susceptibility to the disease. With this marker, we developed a successful eugenic selection procedure for zebu Brahman cattle in Martinique (FWI). Over a period of five years, a marked reduction in disease prevalence, from 0.76 to 0.02 was achieved, and this low level has been maintained over the last two years. The selection procedure, based on a genetic marker system targeting the highly polymorphic BoLA locus, eliminates only those individuals which are at the highest risk of contracting the disease. In the present work, we discuss the properties of this system, including the "heterozygote advantage" and the "frequency dependence" theories, and examine their involvement in the biological mechanisms at the host/pathogen interface. We speculate on the exact role of the MHC molecules in the control of the disease, how the natural selection pressure imposed by the pathogens selectively maintains MHC diversity, and how our results can be practically applied for integrated control of dermatophilosis in developing countries.
Assuntos
Infecções por Actinomycetales/veterinária , Doenças dos Bovinos/genética , Antígenos de Histocompatibilidade Classe II/genética , Seleção Genética , Infecções por Actinomycetales/genética , Infecções por Actinomycetales/imunologia , Alelos , Sequência de Aminoácidos , Animais , Bovinos , Doenças dos Bovinos/imunologia , Primers do DNA/genética , Suscetibilidade a Doenças/veterinária , Eugenia (Ciência) , Haplótipos , HeterozigotoRESUMO
Empirical approaches have guided the development of bacterial cultures. The availability of sequenced genomes now provides opportunities to define culture media for growth of fastidious pathogens with computer modelling of metabolic networks. A key issue is the possibility of growing host-dependent bacteria in cell-free conditions. The sequenced Tropheryma whipplei genome was analysed to identify specific metabolic deficiencies. We used this information to design a comprehensive medium that allowed three established T whipplei strains from culture with human cells and one new strain from a clinical sample to grow axenically. Genomic information can, therefore, provide sufficient clues for designing axenic media for fastidious and uncultured pathogens.