Production of interferon-gamma, interleukin-6, and interleukin-1ß by human peripheral blood mononuclear cells stimulated with novel lys-gingipain synthetic peptides.

BACKGROUND: Periodontitis is a progressive inflammatory process, and its pathogenesis is related to the presence of a dysbiotic subgingival biofilm that elicits the immune response. Porphyromonas gingivalis is a keystone pathogen, and its Lys-gingipain (Kgp) virulence factor is involved in the pathogen-host interaction through the production of cytokines by host cells, but the specific mechanisms of this interaction have not been elucidated. The present study evaluated the in vitro production of interferon-gamma (IFN-γ), interleukin (IL)-6, and IL-1ß cytokines in response to antigenic stimulation of peripheral blood mononuclear cells (PBMCs) with novel Kgp synthetic peptides. METHODS: Our previous in silico study predicted 16 immunogenic peptides from Kgp protein. Nine peptides derived from different regions of the protein were chemically synthesized. The synthetic peptides Kgp12, 17, and 18 were selected based on the immunoglobulin G immunoreactivity in the serum of patients with periodontitis (P) and individuals without periodontitis (WP), and they were used in in vitro stimulation of PBMC derived from groups P and WP. Enzyme-linked immunosorbent assay and microsphere-based flow cytometric assay were used to verify the levels of the cytokines produced in PBMC cultures after 48 hours. RESULTS: Kgp12, 17, and 18 peptides induced lower production of IFN-γ. Kgp12 induced higher levels of IFN-γ in WP than in P individuals. Kgp12 induced higher production of IL-6 and IL-1ß compared with the other stimuli. CONCLUSION: The novel Kgp synthetic peptides tested herein are immunogenic peptides (epitopes) since they induced the production of cytokines by PBMC and therefore may be useful tools in evaluating the pathogen-host interaction.

Infecção por Corynebacterium pseudotuberculosis em equinos: aspectos microbiológicos, clínicos e preventivos / Corynebacterium pseudotuberculosis infection in horses: clinical, microbiological and prevention aspects

Corynebacterium pseudotuberculosis é o agente causador da linfadenite caseosa em caprinos e ovinos, sendo responsável por significativas perdas econômicas na ovinocaprinocultura mundialmente. Esta bactéria Gram-positiva também infecta equinos, causando desde quadros assintomáticos até infecções sistêmicas, podendo levar o animal a óbito. Especificamente no Brasil, não foram relatados casos de infecção em equinos, mas acredita-se que, devido à convivência de pequenos ruminantes infectados com equinos em diversas propriedades rurais, seja natural que ocorra a infecção desses animais. A presente revisão tem como objetivo fornecer informações sobre a bactéria C. pseudotuberculosis, sobre os aspectos epidemiológicos e clínicos da infecção em equídeos, bem como sobre técnicas de manejo para sua prevenção...

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis in sheep and goats, an infectious disease that is responsible for significant economic losses in small ruminants breeding units worldwide. This Gram-positive bacterium can infect horses, causing symptomatic disease to systemic infections, which can lead to animals' death. Specifically in Brazil, there are no scientific records of equine infections, but it is believed that, due to the maintenance of infected small ruminants in close association to horses in many properties, the infection of horses may be a reality. The present work had the objective to present information on the bacteria C. pseudotuberculosis, on the epidemiological and clinical aspects of the infection in horses, as well as information about breeding procedures that can be adopted in order to prevent the infection...

Development of an indirect ELISA to detect Corynebacterium pseudotuberculosis specific antibodies in sheep employing T1 strain culture supernatant as antigen / Desenvolvimento de ELISA indireto para detectar anticorpos específicos contra Corynebacterium pseudotuberculosis em ovinos utilizando o supernadante de cultura da cepa T1 como antígeno

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA), a chronic disease that affects goats and sheep, characterized by granuloma formation in subcutaneous and internal lymph nodes. CLA causes significant economic losses to commercial goat herds. In this study, we aimed to test secreted antigens secreted from T1 strain bacteria grown in brain heart infusion (BHI) broth in an indirect ELISA system to determine the presence of specific immunoglobulins against C. pseudotuberculosis. We analyzed the BHI antigen electrophoretic profile and the recognition pattern by infected sheep sera samples. The ELISA results were compared with multiplex PCR assay and IFN-gamma production. The ELISA was able to discriminate between negative and positive animals, with a sensitivity of 89% and a specificity of 99%, using microbiological isolation as gold standard. When this assay was compared with multiplex PCR and specific IFN-gamma quantification, six discrepant results were found among thirty-two samples. We concluded that the ELISA using antigens secreted from C. pseudotuberculosis T1 strain growth in BHI broth culture can be used for the serodiagnosis of CLA in sheep.

Corynebacterium pseudotuberculosis é o agente etiológico da linfadenite caseosa (LC) uma doença crônica que afeta ovinos e caprinos caracterizada pela formação de granulomas em linfonodos. A LC causa perdas econômicas significativas em criações de pequenos ruminantes. Este trabalho teve como objetivo testar antígenos secretados da cepa T1 da bactéria em um sistema de ELISA indireto para detecção de anticorpos específicos contra C. pseudotuberculosis. O perfil eletroforético do antígeno foi analisado, bem como o padrão de reconhecimento por soros de animais infectados. Os resultados do ELISA foram comparados com ensaio de multiplex PCR e com teste de indução de produção específica de IFN-gama. O ELISA foi capaz de discriminar animais positivos de animais negativos, com sensibilidade de 89% e especificidade de 99%, usando o isolamento microbiológico como padrão ouro. Quando o ensaio foi comparado com o multiplex PCR e a produção específica de IFN-gama, somente seis resultados discrepantes foram encontrados em trinta e duas amostras. Pode-se concluir que o ensaio de ELISA desenvolvido pode ser utilizado com alto grau de confiança para o diagnóstico da LC em ovinos.