RESUMO
This work evaluated aspects of the immune response of BALB/c mice infected with Corynebacterium pseudotuberculosis (T1 and C57). The fifteen BALB/c mice were euthanized after 70 days of infection and morphologically evaluated, also analyzing the innate and adaptive immune responses. The C57 strain induced more pronounced morphological changes than the T1 strain. There was an increase in CD4+ and CD8+ T cells identified during infection with the C57 strain. Cytokines of the inflammatory profile IL-1α and IL-6 and regulatory IL-13 and IL-10 presented significant differences. Cytokines IL-2, IL-4, INF-γ, IL-22, IL-21, and IL-27 did not differ significantly between groups. The obtained results contribute to a better understanding of the type of response and the immunological mechanisms involved during infection with different strains of C. pseudotuberculosis.
Assuntos
Linfócitos T CD8-Positivos , Infecções por Corynebacterium , Corynebacterium pseudotuberculosis , Citocinas , Camundongos Endogâmicos BALB C , Animais , Corynebacterium pseudotuberculosis/imunologia , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Camundongos , Citocinas/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD4-Positivos/imunologia , Interleucina-10 , Imunidade Adaptativa , Imunidade Inata , Interleucina-6 , Modelos Animais de Doenças , Camundongos Endogâmicos C57BL , Interleucina-1alfa/metabolismo , Interleucina-1alfa/imunologia , Interferon gama/metabolismo , Interleucina-4/metabolismo , Interleucinas , Interleucina-2/metabolismoRESUMO
For a long time, the scientific community has described the need for a continued update in practices that ensure the welfare of animals undergoing experimentation. In addition to approaches on principles of care and use of animals, there is a more current emerging concern: defining an appropriate end point in experiments that use animals for research, teaching and testing. The term "endpoint" is defined as the point at which an experimental animal's pain and/or distress is terminated, minimized, or reduced humanely. In the present study, we established an endpoint in Balb/C mice for caseous lymphadenitis vaccine trials, which can be considered as a highly important parameter since several studies are being developed to control the disease efficiently. Mice were monitored daily until the 30th day after infection with pathogenic strain of C. pseudotuberculosis MIC-6 using the most relevant parameters for the appearance of clinical signs of caseous lymphadenitis (CLA), such as abscesses, lethargy, and loss of weight and hair. The endpoint was found to be a weight loss of 0.2167 g after five days or 10% weight loss in less than five days. In conclusion, the findings reported here will help improve animal's well-being during vaccine trials for CLA and consequently represent significant contribution to animal's welfare.
Assuntos
Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/prevenção & controle , Modelos Animais de Doenças , Determinação de Ponto Final/métodos , Linfadenite/prevenção & controle , Redução de Peso , Bem-Estar do Animal , Animais , Infecções por Corynebacterium/imunologia , Corynebacterium pseudotuberculosis/imunologia , Linfadenite/microbiologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Caseous lymphadenitis (CLA) is a chronic disease responsible for significant economic losses in sheep and goat breeding worldwide. The treatment for this disease is not effective, and an intense vaccination schedule would be the best control strategy. In this study, we evaluated the associations of rCP09720 or rCP01850 proteins from Corynebacterium pseudotuberculosis with recombinant exotoxin phospholipase D (rPLD) as subunit vaccines in mice. Four experimental groups (10 animals each) were immunized with a sterile 0.9% saline solution (G1), rPLD (G2), rPLDâ¯+â¯rCP09720 (G3), and rPLDâ¯+â¯rCP01850 (G4). The mice received two doses of each vaccine at a 21-day interval and were challenged 21â¯days after the last immunization. The animals were evaluated daily for 40â¯days after the challenge, and mortality rate was recorded. The total IgG production level increased significantly in the experimental groups on day 42 after the first vaccination. Similarly, higher levels of specific IgG2a were observed in experimental groups G2, G3, and G4 compared to the IgG1 levels on day 42. G4 showed a significant (pâ¯<â¯.05) humoral response against both antigens of the antigenic formulations. The cellular immune response induced by immunization was characterized by a significant (pâ¯<â¯.05) production of interferon-γ compared to that in the control, while the concentrations of interleukin (IL)-4 and IL-12 were not significant in any group. A significant increase of tumor necrosis factor was observed only in G4. The survival rates after the challenge were 30% (rPLD), 40% (rPLDâ¯+â¯rCP09720), and 50% (rPLDâ¯+â¯rCP01850). Thus, the association of rCP01850 with rPLD resulted in the best protection against the challenge with C. pseudotuberculosis and induced a more intense type 1 T-helper cell immune response.
Assuntos
Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/prevenção & controle , Corynebacterium pseudotuberculosis/imunologia , Linfadenite/veterinária , Fosfolipase D/imunologia , Proteínas Recombinantes/imunologia , Fosfatase Ácida/administração & dosagem , Fosfatase Ácida/genética , Fosfatase Ácida/imunologia , Animais , Anticorpos Antibacterianos/sangue , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/genética , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/química , Corynebacterium pseudotuberculosis/enzimologia , Corynebacterium pseudotuberculosis/genética , Esterases/administração & dosagem , Esterases/genética , Esterases/imunologia , Cabras/microbiologia , Imunidade Celular , Imunoglobulina G/sangue , Interferon gama/biossíntese , Interferon gama/imunologia , Linfadenite/imunologia , Linfadenite/microbiologia , Linfadenite/prevenção & controle , Camundongos , Fosfolipase D/administração & dosagem , Fosfolipase D/genética , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Ovinos/microbiologia , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/prevenção & controle , Células Th1/imunologia , Vacinação/veterinária , Vacinas de Subunidades Antigênicas/administração & dosagem , Vacinas de Subunidades Antigênicas/imunologiaRESUMO
BACKGROUND: Caseous lymphadenitis (CLA) is a disease that affects sheep, goats and occasionally humans. The etiologic agent is the Corynebacterium pseudotuberculosis bacillus. The objective of this study was to build a gene expression library from C. pseudotuberculosis and use immunoscreening to identify genes that encode potential antigenic proteins for the development of DNA and subunit vaccines against CLA. RESULTS: A wild strain of C. pseudotuberculosis was used for extraction and partial digestion of genomic DNA. Sequences between 1000 and 5000 base pairs (bp) were excised from the gel, purified, and the digested DNA fragments were joined to bacteriophage vector ZAP Express, packaged into phage and transfected into Escherichia coli. For immunoscreening a positive sheep sera pool and a negative sera pool for CLA were used. Four clones were identified that strongly reacted to sera. The clones were confirmed by polymerase chain reaction (PCR) followed by sequencing for genomic comparison of C. pseudotuberculosis in GenBank. The genes identified were dak2, fagA, fagB, NlpC/P60 protein family and LPxTG putative protein family. CONCLUSION: Proteins of this type can be antigenic which could aid in the development of subunit or DNA vaccines against CLA as well as in the development of serological tests for diagnosis. Immunoscreening of the gene expression library was shown to be a sensitive and efficient technique to identify probable immunodominant genes.
Assuntos
Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/isolamento & purificação , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/genética , Corynebacterium pseudotuberculosis/imunologia , Linfadenite/veterinária , Animais , Antígenos de Bactérias/sangue , Bacteriófagos/genética , Sequência de Bases , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/prevenção & controle , Corynebacterium pseudotuberculosis/patogenicidade , Bases de Dados de Ácidos Nucleicos , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Biblioteca Gênica , Genes Bacterianos/genética , Genoma Bacteriano , Doenças das Cabras/sangue , Doenças das Cabras/imunologia , Doenças das Cabras/microbiologia , Cabras , Linfadenite/imunologia , Linfadenite/microbiologia , Reação em Cadeia da Polimerase/veterinária , Ovinos , Doenças dos Ovinos/sangue , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/microbiologia , Vacinas de DNA/uso terapêuticoRESUMO
PURPOSE: We tested the efficacy of the esterase encoded by cp1002_RS09720 from Corynebacteriumpseudotuberculosis in recombinant subunit and DNA caseous lymphadenitis (CLA) vaccines. This target was predicted as one of the best CLA vaccine candidates by mature epitope density analysis. METHODOLOGY: Gene cp1002_RS09720 was cloned into two different vectors (pAE for subunit vaccine and pTARGET for DNA vaccine). Four groups of 15 mice each were immunized with the recombinant esterase rCP09720 associated with aluminium hydroxide adjuvant (G1), pTARGET/cp09720 DNA vaccine (G2), a naked pTARGET (G3) or PBS as a negative control (G4). Immunization occurred in two doses intercalated by a 21 day interval. Twenty-one days after the last dose administration, animals were challenged with a virulent C. pseudotuberculosis MIC-6 strain. RESULTS: G1 showed high levels of IgG1 and IgG2a on days 21 and 42 post-immunization and a significant level of IFN-γ (P<0.05), suggesting a Th1 response. The protection levels obtained were 58.3 and 16.6â% for G1 and G2, respectively. CONCLUSION: The subunit vaccine composed of the recombinant esterase rCP09720 and Al(OH)3 is a promising antigenic formulation for use against CLA.
Assuntos
Infecções por Corynebacterium/prevenção & controle , Corynebacterium pseudotuberculosis/enzimologia , Corynebacterium pseudotuberculosis/imunologia , Esterases/genética , Linfadenite/prevenção & controle , Vacinas de DNA/imunologia , Adjuvantes Imunológicos , Animais , Infecções por Corynebacterium/imunologia , Corynebacterium pseudotuberculosis/genética , Corynebacterium pseudotuberculosis/patogenicidade , Citocinas/metabolismo , Esterases/administração & dosagem , Esterases/imunologia , Imunoglobulina G/sangue , Interferon gama/imunologia , Linfadenite/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Células Th1/imunologia , Vacinação , Vacinas de DNA/administração & dosagem , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/isolamento & purificaçãoRESUMO
BACKGROUND: Caseous lymphadenitis (CL) is a contagious infectious disease of small ruminants caused by Corynebacterium pseudotuberculosis. Is characterized by the formation of abscesses in the lymph nodes and intestines of infected animals, induced by inflammatory cytokines. The production of cytokines, such as IL-10, TNF-α, IL-4 and IFN-γ, is regulated by mitogen-activated protein kinase (MAPK) pathway activation. The present study investigated the involvement of MAPK pathways (MAPK p38, ERK 1 and ERK 2) with respect to the production of cytokines induced by antigens secreted by C. pseudotuberculosis over a 60-day course of infection. CBA mice (n = 25) were divided into three groups and infected with 102 colony forming units (CFU) of attenuated strain T1, 102 CFU of virulent strain VD57 or sterile saline solution and euthanized after 30 or 60 days. Murine splenocytes were treated with specific inhibitors (MAPK p38 inhibitor, ERK 1/2 inhibitor or ERK 2 inhibitor) and cultured with secreted antigens obtained from pathogenic bacteria (SeT1 or SeVD57). RESULTS: The MAPK pathways evaluated were observed to be involved in the production of IL-10, under stimulation by secreted antigens, while the MAPK p38 and ERK 1 pathways were shown to be primarily involved in TNF-α production. By contrast, no involvement of the MAPK p38 and ERK 1 and 2 pathways was observed in IFN-γ production, while the ERK 2 pathway demonstrated involvement in IL-4 production only in the mouse splenocytes infected with VD57 under stimulation by SeT1. CONCLUSION: The authors hypothesize that MAPK p38 and ERK 1 pathways with respect to TNF-α production, as well as the MAPK p38 and ERK 1 and 2 pathways in relation to IL-10 production under infection by C. pseudotuberculosis are important regulators of cellular response. Additionally, the lack of the MAPK p38 and ERK 1/2 pathways in IFN-γ production in infected CBA murine cells stimulated with the two secreted/excreted antigens, in IL-4 production showing involvement only via the ERK 2 pathway under stimulation by SeT1 antigen during 60-day infection period with the virulent strain, suggests that these pathways regulated the production of pro-inflammatory and regulatory cytokines in the splenic cells of CBA mice.
Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/imunologia , Citocinas/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Transdução de Sinais , Animais , Antígenos de Bactérias/imunologia , Células Cultivadas , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/patologia , Feminino , Leucócitos Mononucleares/imunologia , Masculino , Camundongos Endogâmicos CBA , Baço/imunologiaRESUMO
Corynebacterium bovis is one of the most commonly isolated bacteria from aseptically collected bovine milk samples. The objective of the current study was to characterize the bovine innate immune response by evaluating milk polymorphonuclear neutrophilic leukocytes (PMNL) in mammary glands infected with C. bovis. Twenty quarters infected with C. bovis and 28 culture-negative quarters (with milk somatic cell count <1×10(5) cells/mL) were used. The percentages of milk PMNL and the PMNL expression of L-selectin (CD62L), ß2-integrin (CD11b), and one of the endothelial-selectin ligands (CD44), as well as the levels of intracellular reactive oxygen species (ROS) and the phagocytosis of Staphylococcus aureus, were evaluated by flow cytometry. The apoptosis and necrosis rates of the PMNL were quantified using dual-color flow cytometry with fluorescein-labeled annexin and propidium iodide. The present study revealed a higher percentage of PMNL in the milk from C. bovis-infected quarters, although no significant differences were found in levels of CD44, CD62L, or CD11b expression among the PMNL. A lower percentage of apoptotic PMNL was observed in C. bovis-infected quarters, as well as higher percentages of viable PMNL and of PMNL that produced intracellular ROS. However, no alterations were observed in phagocytosis of Staph. aureus by the PMNL or in intensity of intracellular ROS production by PMNL. Thus, results from this investigation of the PMNL function support, at least in part, the fact that intramammary infections by C. bovis may offer protection against intramammary infections by other bacteria.
Assuntos
Infecções por Corynebacterium/imunologia , Corynebacterium/imunologia , Mastite Bovina/imunologia , Mastite Bovina/microbiologia , Leite/imunologia , Neutrófilos/imunologia , Animais , Antígeno CD11b/imunologia , Bovinos , Separação Celular/veterinária , Corynebacterium/isolamento & purificação , Feminino , Receptores de Hialuronatos/imunologia , Selectina L/imunologia , Leite/citologia , Leite/microbiologia , Neutrófilos/citologia , Fagocitose , Espécies Reativas de Oxigênio/metabolismoRESUMO
During the last decade the majority of diphtheria cases in Europe had Corynebacterium ulcerans as the etiologic agent with dogs and cats as the reservoir hosts. However, little has been documented about the virulence factors of this zoonotic pathogen. To set up an in vivo experimental C. ulcerans infection model, conventional Swiss Webster mice were intravenously infected with different doses (from 1 × 10(7) to 5 × 10(9) bacteria per mouse) of C. ulcerans strains, namely 809 (from human lower respiratory tract), BR-AD22 (from asymptomatic dog nares) and CDC-KC279. Mortality rates were demonstrated by LD(50) values ranging from 1.9 × 10(8) to 1.3 × 10(9). Viable bacteria were recovered from blood, kidneys, liver, spleen and joints. For CDC-KC279 and 809 strains (2 × 10(8)mL(-1)) approximately 85% and 72% of animals with articular lesions were observed, respectively; BR-AD22-infected mice showed no signs of arthritis. CDC-KC279 and 809 strains exhibited higher arthritogenic potential when compared to the homologous toxigenic (ATCC27012) and non-toxigenic (ATCC27010) strains of Corynebacterium diphtheriae. A high number of affected joints and arthritis index in addition to the histopathological features, including subcutaneous edema, inflammatory infiltrate, damage to bone tissue and synoviocyte hypertrophy, indicated a strain-dependent ability of C. ulcerans strains to cause severe polyarthritis. A correlation between the arthritis index and systemic levels of IL-6 and TNF-α was observed for C. ulcerans strains, with the exception of the non-arthritogenic BR-AD22 strain. In conclusion, C. ulcerans revealed a strain-dependent arthritogenic potential independent of DNAse, PLD and diphtheria toxin production.
Assuntos
Artrite Infecciosa/microbiologia , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/patologia , Corynebacterium/fisiologia , Animais , Artrite Infecciosa/patologia , Carga Bacteriana , Corynebacterium/imunologia , Infecções por Corynebacterium/imunologia , Corynebacterium diphtheriae/fisiologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Masculino , Camundongos , Especificidade da Espécie , Fatores de TempoRESUMO
The aim of this study was to evaluate an indirect specific ELISA developed for the detection of humoral immune response in vaccinated sheep and/or challenged with a Corynebacterium pseudotuberculosis strain. Healthy 4 month-old lambs were distributed into 4 groups: Group 1 immunized (G1, n = 5), Group 2 vaccinated/inoculated (G2, n = 8), Group 3 inoculated (G3, n = 2) and Group 4 control (G4, n = 2). Groups G1 and G2 received two doses of an experimental bacterin. Four weeks postvaccination, G2 and G3 groups were challenged with a C. pseudotuberculosis strain. Serological titers were studied by ELISA for 7 months and pathological studies were performed in groups G2, G3 and G4 by taking lung and lymph node samples for bacteriology and histopathology. The inoculated strain in G2 and G3 animals reproduced the macroscopic and microscopic lesions typical of caseous lymphadenitis (CL) and was isolated from the inoculation site, lymph nodes and/or lung in 7/8 animals from G2, and 2/2 animals of G3. The developed ELISA test had sensitivity and specificity of 98% and 100% respectively, detected significant differences between serological reactors of different experimental groups and allowed to establish a relationship with the type of treatment. We conclude that the developed ELISA may be a useful tool to identify infected animals with positive clinical CL.
Assuntos
Anticorpos Antibacterianos/análise , Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Linfadenite/veterinária , Doenças dos Ovinos/imunologia , Ovinos/imunologia , Vacinação/veterinária , Animais , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/prevenção & controle , Ensaio de Imunoadsorção Enzimática/métodos , Pulmão/imunologia , Linfonodos/imunologia , Linfadenite/imunologia , Linfadenite/microbiologia , Linfadenite/prevenção & controle , Distribuição Aleatória , Sensibilidade e Especificidade , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/prevenção & controleRESUMO
El objetivo de este trabajo fue evaluar un ELISA indirecto desarrollado para medir la respuesta inmune humoral en carneros vacunados contra la linfoadenitis caseosa (LC) y/o desafiados con una cepa de Corynebacterium pseudotuberculosis homóloga. Se distribuyeron corderos de 4 meses clínicamente sanos en 4 grupos: grupo 1, corderos vacunados (G1, n = 5); grupo 2, corderos vacunados e inoculados (G2, n = 8); grupo 3, corderos inoculados (G3, n = 2); y grupo 4, control (G4, n = 2). Los animales del G1 y del G2 recibieron dos dosis de una bacterina experimental; los del G2 y del G3 fueron desafiados con una cepa de C. pseudotuberculosis cuatro semanas posvacunación. Se estudiaron por ELISA los títulos serológicos durante 7 meses y se efectuaron las necropsias en los grupos G2, G3 y G4. Se tomaron muestras de pulmón y linfonódulos para efectuar estudios bacteriológicos e histopatológicos. La cepa inoculada en los animales del G2 y del G3 reprodujo las lesiones macroscópicas y microscópicas típicas de la LC; ésta fue aislada del sitio de inoculación, de linfonódulos o de pulmón en 7/8 animales del G2 y en 2/2 animales del G3. La prueba de ELISA, con una sensibilidad del 98% y una especificidad del 100%, detectó diferencias significativas entre los serorreactores de los diferentes grupos experimentales y permitió establecer una relación con el tipo de tratamiento aplicado. Se concluye que el ELISA desarrollado puede ser una herramienta útil para identificar animales infectados y con clínica positiva a la LC.
The aim of this study was to evaluate an indirect specific ELISA developed for the detection of humoral immune response in vaccinated sheep and/or challenged with a Corynebacterium pseudotuberculosis strain. Healthy 4 month-old lambs were distributed into 4 groups: Group 1 immunized (G1, n = 5), Group 2 vaccinated/inoculated (G2, n = 8), Group 3 inoculated (G3, n = 2) and Group 4 control (G4, n = 2). Groups G1 and G2 received two doses of an experimental bacterin. Four weeks postvaccination, G2 and G3 groups were challenged with a C. pseudotuberculosis strain. Serological titers were studied by ELISA for 7 months and pathological studies were performed in groups G2, G3 and G4 by taking lung and lymph node samples for bacteriology and histopathology. The inoculated strain in G2 and G3 animals reproduced the macroscopic and microscopic lesions typical of caseous lymphadenitis (CL) and was isolated from the inoculation site, lymph nodes and/or lung in 7/8 animals from G2, and 2/2 animals of G3. The developed ELISA test had sensitivity and specificity of 98% and 100% respectively, detected significant differences between serological reactors of different experimental groups and allowed to establish a relationship with the type of treatment. We conclude that the developed ELISA may be a useful tool to identify infected animals with positive clinical CL.
Assuntos
Animais , Anticorpos Antibacterianos/análise , Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Linfadenite/veterinária , Doenças dos Ovinos/imunologia , Ovinos/imunologia , Vacinação/veterinária , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/prevenção & controle , Ensaio de Imunoadsorção Enzimática/métodos , Pulmão/imunologia , Linfonodos/imunologia , Linfadenite/imunologia , Linfadenite/microbiologia , Linfadenite/prevenção & controle , Distribuição Aleatória , Sensibilidade e Especificidade , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/prevenção & controleRESUMO
Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a disease that affects goats and sheep, and can cause severe economic losses. In this study, four different antigenic extracts were obtained from the attenuated strain T1, which was isolated in the state of Bahia (Brazil). Forty-four Canindé breed goats were divided in five groups, each receiving a different antigen solution and saline buffer as a control. The humoral response was monitored through the identification of specific IgG by indirect ELISA and Western Blotting, and the production of IFN-gamma was followed in order to observe the activation of cellular response. After twelve weeks of antigen inoculation, the animals were challenged with 2 x 10(5)CFU of a wild strain, also isolated in Bahia, and necropsy was performed on all animals twelve weeks afterwards. It was observed that the attenuated bacteria gave a protection of 33.3%, in addition to the weak humoral response elicited. Animals inoculated with secreted antigen associated with Freund's incomplete adjuvant and oligodeoxynucleotide containing unmethylated CpG dinucleotides (CpG ODN) showed a strong humoral response, but this inoculation could not prevent the spread of challenge bacteria in the majority of animals. These results demonstrate the immunogenic potential of the attenuated T1 strain in the development of a vaccine against caseous lymphadenitis in goats.
Assuntos
Antígenos de Bactérias/imunologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis , Doenças das Cabras/prevenção & controle , Linfadenite/veterinária , Animais , Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/sangue , Infecções por Corynebacterium/imunologia , Doenças das Cabras/sangue , Doenças das Cabras/imunologia , Cabras , Immunoblotting , Imunoglobulina G/sangue , Interferon gama/sangue , Linfadenite/prevenção & controleRESUMO
Corynebacterium pseudotuberculosis, a Gram-positive intracellular pathogen, is the etiological agent of caseous lymphadenitis or CLA. This bacterium infects goats and sheep and causes great economic losses worldwide annually, mainly for goat producers. Despite its importance, CLA is still poorly characterized. However, with advances in the genomic field, many C. pseudotuberculosis genes have already been characterized, mainly those related to virulence such as phospholipase D. Here, we examined the use of the several available genes of C. pseudotuberculosis and reviewed their applications in vaccine construction, more efficient diagnostics for CLA, and control of this disease, among other applications.
Assuntos
Proteínas de Bactérias/genética , Infecções por Corynebacterium/diagnóstico , Corynebacterium pseudotuberculosis/genética , Animais , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/genética , Vacinas Bacterianas/imunologia , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/imunologia , Corynebacterium pseudotuberculosis/patogenicidade , Linfadenite/diagnóstico , Linfadenite/imunologia , Linfadenite/microbiologia , Virulência/genéticaRESUMO
Corynebacterium pseudotuberculosis, a Gram-positive intracellular pathogen, is the etiological agent of caseous lymphadenitis or CLA. This bacterium infects goats and sheep and causes great economic losses worldwide annually, mainly for goat producers. Despite its importance, CLA is still poorly characterized. However, with advances in the genomic field, many C. pseudotuberculosis genes have already been characterized, mainly those related to virulence such as phospholipase D. Here, we examined the use of the several available genes of C. pseudotuberculosis and reviewed their applications in vaccine construction, more efficient diagnostics for CLA, and control of this disease, among other applications.
Assuntos
Animais , Corynebacterium pseudotuberculosis/genética , Infecções por Corynebacterium/diagnóstico , Corynebacterium pseudotuberculosis/imunologia , Corynebacterium pseudotuberculosis/patogenicidade , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/microbiologia , Vacinas Bacterianas/genética , Vacinas Bacterianas/imunologia , Virulência/genéticaRESUMO
Corynebacterium pseudotuberculosis is the causal agent of caseous lymphadenitis, a chronic illness that attacks goats and sheep characterized by pyogranulomas formation in lymph nodes and organs. Regarding the current knowledge of the pathogenesis of the caseous lymphadenitis, there is evidence that besides the humoral response the induction of a durable cellular response is fundamental for its control. In this sense, research on antigens of C. pseudotuberculosis that are capable to inducing cellular immunity is an important step for the development of diagnosis tests and more efficient vaccines. In the present study, the interferon-gamma production in cultures of whole blood from infected goats stimulated with secreted bacterial antigen or somatic antigen were used to evaluate the cellular response. The results demonstrated a significant difference in the ability of the two antigens to induce a cellular response. That is, IFN-gamma production was high with cells from infected animals in response to the secreted antigen while IFN-gamma production was low when somatic antigen was used. The concomitant use of these antigens with PWM also showed differences. That is, the secreted antigen increased the IFN-gamma production induced by PWM, while the somatic antigen seems not to have altered the response to PWM.
Assuntos
Antígenos de Bactérias/administração & dosagem , Corynebacterium pseudotuberculosis/imunologia , Cabras/imunologia , Interferon gama/biossíntese , Animais , Células Sanguíneas/imunologia , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/patogenicidade , Feminino , Doenças das Cabras/imunologia , Cabras/microbiologia , Técnicas In Vitro , Linfadenite/imunologia , Linfadenite/veterinária , Masculino , Mitógenos de Phytolacca americana/administração & dosagemRESUMO
Corynebacterium pseudotuberculosis is the cause of caseous lymphadenitis (CLA) in small ruminants, a chronic granulomatous disease that provokes significant zootechnics losses to ovine and goat breeders in northern Brazil. The present work was conducted to analyse aspects of humoral and cellular immune responses after experimental infection. Eight goats were infected intradermally with a single dose of virulent C. pseudotuberculosis strain and specific IgG, interferon-gamma (IFN-gamma) production as well as IgG avidity and antigens pattern recognition dynamics against an excreted-secreted antigen were recorded during 20 weeks. At the end of the follow-up, animals were slaughtered and necropsied. Although no animals showed apparent clinical signs of infection at the end of the trial, IFN-gamma response, even more so than the humoral response, differentiated animals into two groups of high or medium/low response. The time-course of IFN-gamma production presented a short-lived primary response on day 5 after infection of animals of both groups, and a strong and long lasting secondary response starting on day 16 after infection in the high response group. The indirect ELISA used was able to detect a positive antibody titre between 6 and 11 days after infection in the two groups. IgG avidity index oscillated initially between 15 and 45%, and showed approximately 5% units increment during the 20 follow-up weeks. With only one individual exception, the qualitative antigens pattern recognition showed on day 11 after infection remained constant through the experiment. IgG avidity is highly correlated with IgG production, but could not be related with specific immunodominant bands. Both humoral and cellular responses kinetics presented a similar pattern of activation/deactivation but necropsy results suggested that the IFN-gamma test would be a very specific marker of CLA status.
Assuntos
Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/imunologia , Doenças das Cabras/imunologia , Imunoglobulina G/biossíntese , Interferon gama/biossíntese , Linfadenite/veterinária , Animais , Anticorpos Antibacterianos/sangue , Afinidade de Anticorpos/imunologia , Antígenos de Bactérias/imunologia , Western Blotting/veterinária , Brasil , Infecções por Corynebacterium/imunologia , Infecções por Corynebacterium/patologia , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Doenças das Cabras/microbiologia , Cabras , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Interferon gama/sangue , Cinética , Linfadenite/imunologia , Linfadenite/patologia , MasculinoRESUMO
In 1999, a case of diphtheria in a 32-year-old woman was reported. The patient developed a sore throat immediately after participating of a five-day meeting with European workers in Rio de Janeiro. Her history included complete pediatric immunization (DTP) and three doses of adult formulation tetanus and diphtheria toxoid (dT) two years earlier. Clinical diagnosis of diphtheria was not made until microbiologic examination of specimens confirmed toxigenicity of Corynebacterium diphtheriae var. gravis, a biotype currently found circulating within Europe where diphtheria remains epidemic. This case reinforces the potential susceptibility of Brazilian adults to epidemic diphtheria in the vaccine era.