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Helton da Costa Santiago5170293352882261Lis Ribeiro do Valle AntonelliMauro Martins TeixeiraAna Maria Caetano de FariaAndréa Teixeira de CarvalhoMarcelo Torres Bozza4369104520287380Marcela Helena Gonçalves Pereira de Oliveira2023-06-28T17:22:44Z2023-06-28T17:22:44Z2020-03-30http://hdl.handle.net/1843/55490Introdução: A dengue é uma infecção viral e pode ser classificada em dengue leve, normalmente tratada domiciliarmente, e dengue com sinais de alarme (sa+) e dengue grave (sa+/grave), que requerem hospitalização. Manifestações graves da dengue são consideradas o resultado da ativação exacerbada da resposta imune. A importância dos mecanismos reguladores promovidos por moléculas e citocinas anti-inflamatórias das células T multifuncionais e T reguladoras (Tregs) no controle das respostas inflamatórias é evidente em muitas doenças infecciosas. Portanto, nosso objetivo foi investigar a presença destes mecanismos em diferentes formas clínicas da infecção por dengue. Métodos e Resultados: células mononucleares do sangue periférico (PBMCs) de pacientes com dengue foram cultivadas na presença da biblioteca de peptídeos para as proteínas virais de envelope (ENV) ou NS3 de DENV1. Pacientes com dengue leve apresentaram níveis mais altos de IFNγ, TNF e IL12p70 no plasma quando comparado com o grupo sa+/grave. As frequências de células T CD4+ ou CD8+ específicas para DENV simples ou duplo produtoras (TNF ou IL10 ou IFNγ/TNF ou IFNγ/IL10) foram maiores em dengue leve quando comparado com as formas clínicas sa+/grave. Além disso, pacientes com dengue leve apresentaram níveis mais altos de células T específicas para NS3 triplo produtoras de IFNγ, TNF e IL10 que o grupo sa+/grave. Dentre as moléculas reguladoras avaliadas, verificamos que pacientes com dengue aumentam as frequências de células Teff GITR+ ou LAP+ ou PD1+, e células Tregs CD226+ e CTLA4+. Por outro lado, pacientes com dengue leve apresentaram elevadas frequências de Tregs CD200+ e de modo importante elevadas frequências de tTregs GITR+ produtoras de IL10 específicas ao DENV, população pouco entrada em pacientes com dengue sa+/grave. Além disso, usando o mapeamento de epítopo da biblioteca de peptídeos do ENV de DENV1, conseguimos identificar peptídeos associados a produção de IL10 pelas Tregs. Conclusão: A evolução clínica da dengue parece relacionar-se com mecanismos reguladores da resposta imune. Tregs de indivíduos com dengue sa+/grave apresentaram deficiência importante em marcadores de ativação, sugerindo um fenótipo disfuncional e a produção de IL10 por células T multifuncionais e por células tTregs GITR+ está associada à apresentação clínica de dengue leve, sugerindo que esses mecanismos reguladores são importantes para limitar a imunopatologia da dengue.Introduction: Dengue is a viral infection and can be classified into mild dengue, usually treated at home, and dengue with warning signs (sa+) and severe dengue (sa+/severe), which require hospitalization. Severe manifestations of dengue are considered to be the result of exacerbated activation of the immune response. The importance of regulatory mechanisms promoted by anti-inflammatory molecules and cytokines of multifunctional and regulatory T cells (Tregs) in controlling inflammatory responses is evident in many infectious diseases. Therefore, our objective was to investigate the presence of these mechanisms in different clinical forms of dengue infection. Methods and Results: Peripheral blood mononuclear cells (PBMCs) from dengue patients were cultured in the presence of the peptide library for the viral envelope proteins (ENV) or NS3 of DENV1. Patients with mild dengue had higher plasma levels of IFNγ, TNF and IL12p70 when compared to the sa+/severe group. The frequencies of specific CD4+ or CD8+ T cells producing single or double DENV (TNF or IL10 or IFNγ/TNF or IFNγ/IL10) were higher in mild dengue when compared to sa+/severe clinical forms. Furthermore, patients with mild dengue had higher levels of triple NS3-specific T cells producing IFNγ, TNF, and IL10 than the sa+/severe group. Among the regulatory molecules evaluated, we verified that patients with dengue increase the frequencies of Teff GITR+ or LAP+ or PD1+ cells, and Tregs CD226+ and CTLA4+ cells. On the other hand, patients with mild dengue showed high frequencies of CD200+ Tregs and, importantly, high frequencies of GITR+ tTregs that produce IL10 specific to DENV, a population that is rarely used in patients with sa+/severe dengue. Furthermore, using epitope mapping of the DENV1 ENV peptide library, we were able to identify peptides associated with IL10 production by Tregs. Conclusion: The clinical evolution of dengue seems to be related to regulatory mechanisms of the immune response. Tregs from individuals with sa+/severe dengue showed an important deficiency in activation markers, suggesting a dysfunctional phenotype, and the production of IL10 by multifunctional T cells and by tTregs GITR+ cells is associated with the clinical presentation of mild dengue, suggesting that these regulatory mechanisms are important to limit dengue immunopathology.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Bioquímica e ImunologiaUFMGBrasilICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIABioquímica e imunologiaDengueLinfócitos T reguladorescélula TregimurregulaçãodengueMecanismos de regulação da resposta imune na dengue humanainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALTese Final revisada MHGPO.pdfTese Final revisada MHGPO.pdfapplication/pdf5286775https://repositorio.ufmg.br/bitstream/1843/55490/1/Tese%20Final%20revisada%20MHGPO.pdf5bbceb0424bbf4ccf503b18acb56e50aMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/55490/2/license.txtcda590c95a0b51b4d15f60c9642ca272MD521843/554902023-06-28 14:22:45.251oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-06-28T17:22:45Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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