| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1297-1 | ||||
Resumo:Introduction. Ureaplasma diversum infects cattle and plays a significant role in economic losses in the agricultural sector due its association with the development of reproductive and respiratory disorders in these animals. Studies have proposed that membrane-associated lipoproteins (LAMPs) are strongly related to the pathogenicity of this bacteria. Thus, this study aimed to express in the heterologous system the lipoprotein GUDIV-517 from U. diversum (rGUDIV-517) and evaluate the immunogenicity of this antigen in cultured bovine peripheral blood mononuclear cells (PBMC). Material and methods. The vector pET-28a (+) containing the gene sequence gudiv-517 was expressed in Escherichia coli BL21(DE3). Recombinant protein was inoculated into mice (BALB/c) to produce anti-rGUDIV-517 antibodies. Immunogenicity was verified by indirect ELISA. Quantification of nitric oxide (NO) and Hydrogen peroxide (H2O2) was performed in PBMC culture supernatant and expression analysis of IL-1β, TNF-α, TLR2, TLR4, iNOS, and caspase-3 genes were performed by qPCR. Results. This study demonstrated that booster doses of rGUDIV-517 induces increased IgG production and avidity. Recombinant GUDIV-517 also caused upregulation of immune response with increased of H2O2 and NO production and expression pro-inflammatory markers in PBMC culture compared to the negative control. Characterization of the antigenicity of the recombinant proteins revealed that they induce the production of avid antibodies. Stimulation of the expression of proinflammatory cytokines and the production of NO and H2O2 indicates the potential for immunobiological applications of rGUDIV-517. Conclusion. The results of this study justify further investigation into challenge experiments in cattle. Data from this study show that rGUDIV-517 is a potential antigen for studies aimed at developing immunodiagnostic and prophylactic tests against U. diversum. This would be an efficient method for field diagnosis and, consequently, would help control infections by U. diversum. Palavras-chave: Immunogenicity, lipoprotein, Ureaplasma diversum Agência de fomento:Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) | |||||