RESUMO
Caseous lymphadenitis (CLA) is an infectious disease caused by Corynebacterium pseudotuberculosis in small ruminants and is characterized by the development of granulomas in the lymph nodes, spleen, liver, and lungs. Although little is known about the host-pathogen relationship of this bacterium, it was previously reported that the pathogen's lipids are important for its taxonomic classification and survival inside macrophages. However, there are no studies regarding the composition of these molecules. In this study, cell wall glycolipids from two C. pseudotuberculosis strains presenting different virulence profiles were purified and its composition was characterized. A difference was observed between the electrophoretic and chromatogram profiles for cell wall components from the two strains, mainly among molecules with low molecular weights. IgM from sheep with acute CLA recognized antigens with an estimated molecular weight of 11 kDa of the low-pathogenicity strain, while low-molecular weight antigens from the high-pathogenicity strain presented a lower recognition by these antibodies. Mass spectrometry analysis showed that the cell wall of the high-pathogenicity strain contained glycolipids with high amounts of unsaturated fatty acids and glycerophosphoinositols, which may contribute to the capacity of this strain to cause severe disease. In conclusion, it is indicated that cell wall non-protein antigens can play a key role in C. pseudotuberculosis virulence.
Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Parede Celular/química , Corynebacterium pseudotuberculosis/química , Glicolipídeos/imunologia , Linfadenite/veterinária , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/química , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/imunologia , Corynebacterium pseudotuberculosis/patogenicidade , Glicolipídeos/química , Doenças das Cabras/imunologia , Doenças das Cabras/microbiologia , Cabras/microbiologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Linfonodos/imunologia , Linfonodos/microbiologia , Linfadenite/imunologia , Linfadenite/microbiologia , Ovinos , Doenças dos Ovinos/microbiologia , VirulênciaRESUMO
The target cp1002_RS01850 from Corynebacterium pseudotuberculosis was used to construct a DNA and recombinant subunit vaccine against caseous lymphadenitis. Recombinant protein rCP01850 was expressed in Escherichia coli using pAE vector, and DNA vaccine was engineered with pTARGET vector. BALB/c mice were divided in five groups containing eight animals each, inoculated with: pTARGET/cp01850 as DNA vaccine (G1); rCP01850 plus Al (OH)3 as recombinant subunit vaccine (G2); pTARGET/cp01850 and a boost with rCP01850 plus Al (OH)3 (G3); pTARGET (G4); or Al (OH)3 (G5). Mice were inoculated and blood samples were collected on days 0, 21, and 42 for the analysis of total IgG, IgG1 and IgG2a by ELISA. In each group, five animals were challenged with Mic-6 C. pseudotuberculosis strain, and three were used for cytokine quantification by qPCR. Although no group has been protected by vaccines against lethal challenge, G2 showed an increase in the survival rate after challenge. Significantly higher levels of IL-4, IL-12, IFN-γ, total IgG, IgG1 and IgG2a were also detected for G2, evidencing a mixed Th1/Th2 immunological profile. In conclusion, despite no protection level provided by different vaccinal strategies using cp1002_RS01850 from C. pseudotuberculosis, G2 developed a Th1/Th2 immune response with an increase in survival rate.(AU)
O alvo cp1002_RS01850 de Corynebacterium pseudotuberculosis foi utilizado para construir uma vacina recombinante de subunidade e de DNA contra a linfadenite caseosa. A proteína recombinante rCP01850 foi expressa em Escherichia coli usando o vetor pAE, e a vacina de DNA foi construída com o vetor pTARGET. Camundongos BALB/c foram divididos em grupos de oito animais, inoculados com: pTARGET/cp01850 como vacina de DNA (G1); rCP01850 e Al (OH)3 como vacina recombinante de subunidade (G2); pTARGET/cp01850 e um boost com rCP01850 e Al (OH)3 (G3); pTARGET (G4); ou Al (OH)3 (G5). Os animais foram inoculados e amostras de sangue foram coletadas nos dias 0, 21, e 42 do experimento para a análise de IgG total, IgG1 e IgG2a por ELISA. De cada grupo, cinco animais foram desafiados com a cepa Mic-6 de C. pseudotuberculosis, e três foram usados para a quantificação de citocinas por qPCR. Apesar de nenhum grupo ter sido protegido pelas vacinas testadas contra o desafio letal, G2 apresentou taxa de sobrevida e níveis de IL-4, IL-12, IFN-γ, IgG total, IgG1 e IgG2a significativamente mais altos, evidenciando um perfil imunológico misto Th1/Th2. Conclui-se que apesar das diferentes estratégias vacinais utilizando cp1002_RS01850 de C. pseudotuberculosis não terem sido capazes de gerar proteção, G2 desenvolveu uma resposta Th1/Th2 e elevou a taxa de sobrevida.(AU)
Assuntos
Animais , Camundongos , Fosfatase Ácida , Imunização Secundária/veterinária , Corynebacterium pseudotuberculosis , Linfadenite/imunologia , Proteínas Recombinantes , Hidróxido de AlumínioRESUMO
The target cp1002_RS01850 from Corynebacterium pseudotuberculosis was used to construct a DNA and recombinant subunit vaccine against caseous lymphadenitis. Recombinant protein rCP01850 was expressed in Escherichia coli using pAE vector, and DNA vaccine was engineered with pTARGET vector. BALB/c mice were divided in five groups containing eight animals each, inoculated with: pTARGET/cp01850 as DNA vaccine (G1); rCP01850 plus Al (OH)3 as recombinant subunit vaccine (G2); pTARGET/cp01850 and a boost with rCP01850 plus Al (OH)3 (G3); pTARGET (G4); or Al (OH)3 (G5). Mice were inoculated and blood samples were collected on days 0, 21, and 42 for the analysis of total IgG, IgG1 and IgG2a by ELISA. In each group, five animals were challenged with Mic-6 C. pseudotuberculosis strain, and three were used for cytokine quantification by qPCR. Although no group has been protected by vaccines against lethal challenge, G2 showed an increase in the survival rate after challenge. Significantly higher levels of IL-4, IL-12, IFN-γ, total IgG, IgG1 and IgG2a were also detected for G2, evidencing a mixed Th1/Th2 immunological profile. In conclusion, despite no protection level provided by different vaccinal strategies using cp1002_RS01850 from C. pseudotuberculosis, G2 developed a Th1/Th2 immune response with an increase in survival rate.(AU)
O alvo cp1002_RS01850 de Corynebacterium pseudotuberculosis foi utilizado para construir uma vacina recombinante de subunidade e de DNA contra a linfadenite caseosa. A proteína recombinante rCP01850 foi expressa em Escherichia coli usando o vetor pAE, e a vacina de DNA foi construída com o vetor pTARGET. Camundongos BALB/c foram divididos em grupos de oito animais, inoculados com: pTARGET/cp01850 como vacina de DNA (G1); rCP01850 e Al (OH)3 como vacina recombinante de subunidade (G2); pTARGET/cp01850 e um boost com rCP01850 e Al (OH)3 (G3); pTARGET (G4); ou Al (OH)3 (G5). Os animais foram inoculados e amostras de sangue foram coletadas nos dias 0, 21, e 42 do experimento para a análise de IgG total, IgG1 e IgG2a por ELISA. De cada grupo, cinco animais foram desafiados com a cepa Mic-6 de C. pseudotuberculosis, e três foram usados para a quantificação de citocinas por qPCR. Apesar de nenhum grupo ter sido protegido pelas vacinas testadas contra o desafio letal, G2 apresentou taxa de sobrevida e níveis de IL-4, IL-12, IFN-γ, IgG total, IgG1 e IgG2a significativamente mais altos, evidenciando um perfil imunológico misto Th1/Th2. Conclui-se que apesar das diferentes estratégias vacinais utilizando cp1002_RS01850 de C. pseudotuberculosis não terem sido capazes de gerar proteção, G2 desenvolveu uma resposta Th1/Th2 e elevou a taxa de sobrevida.(AU)
Assuntos
Animais , Camundongos , Fosfatase Ácida , Imunização Secundária/veterinária , Corynebacterium pseudotuberculosis , Linfadenite/imunologia , Proteínas Recombinantes , Hidróxido de AlumínioRESUMO
Cryptococcosis is an opportunistic infection caused by the Basidiomycota Cryptococcus neoformans (Cryptococcus gattii), which affects immunosuppressed patients and less frequently immunocompetent patients. Solid-organ transplant recipients are a particularly high-risk group, depending on the net state of immunosuppression. In these patients, the infection usually appears after the first year after transplant, although it may occur earlier in liver transplant recipients. In most cases, the infection is secondary to the reactivation of a latent infection, although it may be due to an unidentified pretransplant infection by primary infection. Less frequently, it may be transmitted by the graft. The lung and central nervous system are most frequently involved. Extrapulmonary involvement is seen in 75% of the cases, and disseminated disease occurs in 61%, with mortality ranging from 17% to 50% when the central nervous system is involved. Here, we report a case of disseminated cryptococcosis (lymphadenitis, meningitis, pulmonary nodules, and possibly sacroiliitis) in a patient after liver transplant, with good clinical and microbiological outcomes and without relapse.
Assuntos
Criptococose/microbiologia , Transplante de Fígado/efeitos adversos , Pneumopatias Fúngicas/microbiologia , Linfadenite/microbiologia , Infecções Oportunistas/microbiologia , Sacroileíte/microbiologia , Adulto , Antifúngicos/uso terapêutico , Criptococose/diagnóstico , Criptococose/tratamento farmacológico , Criptococose/imunologia , Feminino , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/efeitos adversos , Pneumopatias Fúngicas/diagnóstico , Pneumopatias Fúngicas/tratamento farmacológico , Pneumopatias Fúngicas/imunologia , Linfadenite/diagnóstico , Linfadenite/tratamento farmacológico , Linfadenite/imunologia , Meningite Criptocócica/tratamento farmacológico , Meningite Criptocócica/imunologia , Meningite Criptocócica/microbiologia , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/tratamento farmacológico , Infecções Oportunistas/imunologia , Sacroileíte/diagnóstico , Sacroileíte/tratamento farmacológico , Sacroileíte/imunologia , Resultado do TratamentoRESUMO
This consensus statement update reflects our current published knowledge and opinion about clinical signs, pathogenesis, epidemiology, treatment, complications, and control of strangles. This updated statement emphasizes varying presentations in the context of existing underlying immunity and carrier states of strangles in the transmission of disease. The statement redefines the "gold standard" for detection of possible infection and reviews the new technologies available in polymerase chain reaction diagnosis and serology and their use in outbreak control and prevention. We reiterate the importance of judicious use of antibiotics in horses with strangles. This updated consensus statement reviews current vaccine technology and the importance of linking vaccination with currently advocated disease control and prevention programs to facilitate the eradication of endemic infections while safely maintaining herd immunity. Differentiation between immune responses to primary and repeated exposure of subclinically infected animals and responses induced by vaccination is also addressed.
Assuntos
Doenças dos Cavalos/diagnóstico , Linfadenite/diagnóstico , Infecções Estreptocócicas/veterinária , Animais , Consenso , Surtos de Doenças/prevenção & controle , Surtos de Doenças/veterinária , Doenças dos Cavalos/imunologia , Doenças dos Cavalos/prevenção & controle , Doenças dos Cavalos/terapia , Cavalos , Linfadenite/imunologia , Linfadenite/prevenção & controle , Linfadenite/terapia , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/prevenção & controle , Streptococcus equi/imunologia , Vacinação/veterináriaRESUMO
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
Using flow cytometry, we evaluated the frequencies of CD4(+) and CD8(+) T cells and Foxp3(+) regulatory T cells (Tregs) in mononuclear cells in the jejunum, colon, and cervical and mesenteric lymph nodes of dogs naturally infected with Leishmania infantum and in uninfected controls. All infected dogs showed chronic lymphadenitis and enteritis. Despite persistent parasite loads, no erosion or ulcers were evident in the epithelial mucosa. The colon harbored more parasites than the jejunum. Frequencies of total CD4(+), total Foxp3, and CD4(+) Foxp3(+) cells were higher in the jejunum than in the colon. Despite negative enzyme-linked immunosorbent assay (ELISA) serum results for cytokines, levels of interleukin-10 (IL-10), gamma interferon (IFN-γ), transforming growth factor beta (TGF-ß), and tumor necrosis factor alpha (TNF-α) were higher in the jejunum than in the colon for infected dogs. However, IL-4 levels were higher in the colon than in the jejunum for infected dogs. There was no observed correlation between clinical signs and histopathological changes or immunological and parasitological findings in the gastrointestinal tract (GIT) of canines with visceral leishmaniasis. However, distinct segments of the GIT presented different immunological and parasitological responses. The jejunum showed a lower parasite load, with increased frequencies and expression of CD4, Foxp3, and CD8 receptors and IL-10, TGF-ß, IFN-γ, and TNF-α cytokines. The colon showed a higher parasite load, with increasing expression of IL-4. Leishmania infantum infection increased expression of CD4, Foxp3, IL-10, TGF-ß, IFN-γ, and TNF-α and reduced CD8 and IL-4 expression in both the jejunum and the colon.
Assuntos
Colo do Útero/imunologia , Colo/imunologia , Jejuno/imunologia , Leishmania infantum/imunologia , Leishmaniose Visceral/imunologia , Linfonodos/imunologia , Linfócitos T Reguladores/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/microbiologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/microbiologia , Colo do Útero/microbiologia , Colo/microbiologia , Doenças do Cão/imunologia , Doenças do Cão/microbiologia , Cães , Enterite/imunologia , Enterite/microbiologia , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Feminino , Fatores de Transcrição Forkhead/imunologia , Interferon gama/imunologia , Interleucina-10/imunologia , Interleucina-4/imunologia , Jejuno/microbiologia , Leishmaniose Visceral/microbiologia , Linfonodos/microbiologia , Linfadenite/imunologia , Linfadenite/microbiologia , Masculino , Mucosa/imunologia , Mucosa/microbiologia , Carga Parasitária , Fator de Crescimento Transformador beta/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
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
In the current study, the applicability of the quantification of gamma interferon (IFN-γ) levels for the detection of animals infected with Corynebacterium pseudotuberculosis and for determining caseous lymphadenitis (CLA) clinical status was evaluated. Peripheral blood leukocytes were collected from CLA nonendemic areas animals, from CLA seropositive animals without clinical signs of the disease, and from seropositive animals presenting CLA clinical signs. The leukocytes were stimulated with C. pseudotuberculosis-secreted antigens that were concentrated by the three-phase partitioning technique. An ovine IFN-γ enzyme-linked immunosorbent assay was used to quantify IFN-γ production. Goats and sheep with CLA had higher IFN-γ levels than uninfected seronegative animals. Leukocytes from sheep with CLA chronic abscesses produced higher IFN-γ levels when compared with seropositive sheep without CLA clinical signs, but this difference was not significant in goats. The sensitivity of the assay was 55.8% and 56%, whereas the specificity was 100% and 93%, for goats and sheep, respectively. In conclusion, IFN-γ is a potential marker for the determination of CLA infection status in small ruminants; however, further research is needed to improve assay sensitivity.
Assuntos
Corynebacterium pseudotuberculosis/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças das Cabras/microbiologia , Interferon gama/análise , Leucócitos/imunologia , Linfadenite/veterinária , Doenças dos Ovinos/microbiologia , Animais , Antígenos de Bactérias , Corynebacterium pseudotuberculosis/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Doenças das Cabras/sangue , Doenças das Cabras/diagnóstico , Doenças das Cabras/imunologia , Cabras , Linfadenite/diagnóstico , Linfadenite/imunologia , Linfadenite/microbiologia , Reação em Cadeia da Polimerase/veterinária , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/sangue , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/imunologia , Estatísticas não ParamétricasRESUMO
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, 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
Immune reconstitution inflammatory syndrome (IRIS) is an atypical and unexpected reaction related to highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. IRIS includes an atypical response to an opportunistic pathogen (generally Mycobacterium tuberculosis, Mycobacterium avium complex, cytomegalovirus and herpes varicella-zoster), in patients responding to HAART with a reduction of plasma viral load and evidence of immune restoration based on increase of CD4+ T-cell count. We reported a case of a patient with AIDS which, after a first failure of HAART, developed a subcutaneous abscess and supraclavicular lymphadenitis as an expression of IRIS due to Mycobacterium avium complex after starting a second scheme of HAART.
Assuntos
Infecções Oportunistas Relacionadas com a AIDS/etiologia , Abscesso/microbiologia , Terapia Antirretroviral de Alta Atividade/efeitos adversos , Linfadenite/microbiologia , Infecção por Mycobacterium avium-intracellulare/etiologia , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Infecções Oportunistas Relacionadas com a AIDS/tratamento farmacológico , Infecções Oportunistas Relacionadas com a AIDS/imunologia , Abscesso/tratamento farmacológico , Abscesso/imunologia , Adulto , Contagem de Linfócito CD4 , Humanos , Linfadenite/tratamento farmacológico , Linfadenite/imunologia , Masculino , Infecção por Mycobacterium avium-intracellulare/tratamento farmacológico , Infecção por Mycobacterium avium-intracellulare/imunologia , Síndrome de Resposta Inflamatória Sistêmica/tratamento farmacológico , Síndrome de Resposta Inflamatória Sistêmica/imunologia , Carga ViralRESUMO
Immune reconstitution inflammatory syndrome (IRIS) is an atypical and unexpected reaction related to highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. IRIS includes an atypical response to an opportunistic pathogen (generally Mycobacterium tuberculosis, Mycobacterium avium complex, cytomegalovirus and herpes varicella-zoster), in patients responding to HAART with a reduction of plasma viral load and evidence of immune restoration based on increase of CD4+ T-cell count. We reported a case of a patient with AIDS which, after a first failure of HAART, developed a subcutaneous abscess and supraclavicular lymphadenitis as an expression of IRIS due to Mycobacterium avium complex after starting a second scheme of HAART.
El síndrome inflamatorio de reconstitución inmune (SIRI) es una reacción atípica e inesperada relacionada con el tratamiento antirretroviral de gran actividad (TARGA) en pacientes infectados por el virus de la inmunodeficiencia humana (VIH). El SIRI representa una respuesta inflamatoria frente a un patógeno oportunista (generalmente Mycobacterium tuberculosis, Complejo Mycobacterium avium, citomegalovirus y herpes varicela-zóster) en pacientes que responden a la TARGA con una marcada reducción de la carga viral en plasma y evidencia de una recuperación inmunológica expresada por el incremento de los niveles de linfocitos T CD4+. Presentamos el caso de un paciente con síndrome de inmunodeficiencia adquirida que desarrolló un absceso subcutáneo en muslo derecho y una adenitis supraclavicular izquierda como manifestación de SIRI por Complejo Mycobacterium avium luego del inicio de un segundo esquema de TARGA.
Assuntos
Adulto , Humanos , Masculino , Infecções Oportunistas Relacionadas com a AIDS/etiologia , Abscesso/microbiologia , Terapia Antirretroviral de Alta Atividade/efeitos adversos , Linfadenite/microbiologia , Infecção por Mycobacterium avium-intracellulare/etiologia , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Infecções Oportunistas Relacionadas com a AIDS/tratamento farmacológico , Infecções Oportunistas Relacionadas com a AIDS/imunologia , Abscesso/tratamento farmacológico , Abscesso/imunologia , Linfadenite/tratamento farmacológico , Linfadenite/imunologia , Infecção por Mycobacterium avium-intracellulare/tratamento farmacológico , Infecção por Mycobacterium avium-intracellulare/imunologia , Síndrome de Resposta Inflamatória Sistêmica/tratamento farmacológico , Síndrome de Resposta Inflamatória Sistêmica/imunologia , Carga ViralRESUMO
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
OBJECTIVE: To study the serologic profile of several types of test for toxoplasmosis, in order to contribute to the interpretation of antibody kinetics. METHODS: The clinical and serologic features of 120 cases of lymphadenopathy with known time of clinical onset were studied during 18 months postinfection. Antibody kinetics was determined by Sabin-Feldman dye test, complement fixation with light antigen, IgM immunofluorescent antibody test, and IgM immunosorbent agglutination assay (IgM-ISAGA). Cell-mediated immunity was evaluated by the toxoplasmin skin test. RESULTS: Seventy-five female patients aged 11-54 years (median 27 years) and 45 male patients aged 3-59 years (median 17 years) were studied, 85% of whom were under 30 years of age. Cervical lymph nodes were involved throughout, generally on both sides, with more than one affected ganglion group in 88%. The predominant symptom was asthenia (69%), which persisted in some cases for several months. A negative Sabin-Feldman dye test in a lymphadenopathy with more than three weeks' evolution excludes a toxoplasma etiology. A positive Sabin-Feldman dye test with negative IgM-ISAGA almost invariably excludes recent infection. The Sabin-Feldman dye test was positive in 94% of patients with titers higher than 1 : 16 000 within the first three months. The IgM-ISAGA yielded 98% of positive results, of which 94% were high titers. Titers >/= 1 : 160 in the IgM immunofluorescent antibody test and complement fixation were found to be highly indicative of recent infection, since 87% and 91%, respectively, were found within the first three months. A negative skin test plus positive serology values indicates recent infection. CONCLUSION: Our results indicate that estimation of time of infection on the basis of serologic results is improved by the simultaneous application of several tests, and correlates closely with the presence of clinical lymphadenitis.
Assuntos
Linfadenite/imunologia , Linfadenite/fisiopatologia , Toxoplasmose/imunologia , Toxoplasmose/fisiopatologia , Adolescente , Adulto , Fatores Etários , Criança , Pré-Escolar , Testes de Fixação de Complemento , Feminino , Imunofluorescência , Humanos , Soros Imunes/imunologia , Imunoglobulina M/imunologia , Imunoglobulina M/metabolismo , Cinética , Linfadenite/diagnóstico , Masculino , Pessoa de Meia-Idade , Toxoplasmose/diagnósticoAssuntos
Bacteriemia/imunologia , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/microbiologia , Hospedeiro Imunocomprometido , Infecções por Mycobacterium não Tuberculosas/fisiopatologia , Infecções por Mycobacterium não Tuberculosas/imunologia , Infecções por Mycobacterium não Tuberculosas/prevenção & controle , Linfadenite/imunologia , Linfadenite/microbiologia , Microbiologia Ambiental , Mycobacterium avium subsp. paratuberculosis , Mycobacterium/patogenicidade , Pneumopatias/imunologia , Pneumopatias/microbiologia , Úlcera Cutânea/microbiologiaRESUMO
Se analizaron inmunohistológicamente líneas de cultivo celular infectadas con el HHV-6: HSB2-células T inmaduras y HDLM2-células de Hodgkin, así como ganglios linfáticos de pacientes con enfermedad de Hodgkin y linfadenitis de Kikuchi-Fujimoto (LKF) en relación a la expresión de los productos oncógenos/antioncógenos p53, bcl-2, ras y p21WAF. Se comprobó la proliferación celular inmunohistológicamente mediante anticuerpos contra PCNa (antígeno nuclear de proliferación celular) y la apoptosis se investigó en cortes finos de tejido ganglionar, analizando el ADN fragmentado con marcación final in situ. La LKF mostró alta incidencia de focos de células muertas (linfadenitis histiocítica necrozante), mientras que en la enfermedad de Hodgkin se observó proliferación celular. Con las técnicas utilizadas no se logró mostrar diferencias significativas en la expresión de ADN viral no de antígenos en las líneas celulares, ni en las biopsias de enfermedad de Hodgkin y de LKF. Las células HDLM2 con mejor viabilidad posterior a la infección con HHV-6 y un grado de apoptosis inferior, mostraron una expresión de p53 y de PCNA mucho menor que las células HSB2. Las biopsias de LKF no expresaron p53; ras se observó en menores células que en la enfermedad de Hodgkin y la positividad de PCNA fue tres veces mayor en enfermedad de Hodgkin en comparación con LKf. Sin embargo el bcl-2 se observó con mayor frecuencia en LKF que en enfermedad de Hodgkin. Los resultados no son de fácil interpretación; los datos sugieren la implicación de otros factores exógenos (por ejemplo, citoquinas y factores de crecimiento) en el mecanismo regulatorio de la proliferación celular y de la apoptosis, ambas inducidas probablemente por virus