Avaliação de células dendríticas ativadas como tratamento da esporotricose murina em modelo experimental
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Data
2019-11-25
Autores
Jellmayer, Juliana Aparecida [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A esporotricose é uma micose de distribuição universal causada por fungos termodimórficos do complexo de espécies Sporothrix schenckii (S. schenckii). No Brasil, a esporotricose é considerada endêmica, sendo normalmente adquirida pela inoculação acidental do seu agente causal através da pele ou através da transmissão zoonótica por gatos infectados. As formas clínicas podem variar entre cutânea, linfocutânea e sistêmica, esta última sendo mais comumente observada em pacientes imunodeprimidos. A ineficácia do tratamento antifúngico contra esta micose, especialmente em pacientes imunocomprometidos, tem levado à busca de terapias mais eficazes e seguras. Com base em vários estudos que mostram a eficiente utilização de células dendríticas como ferramenta para o desenvolvimento de vacinas contra diferentes fungos, o objetivo deste trabalho foi avaliar a capacidade protetora de células dendríticas derivadas da medula óssea (BMDCs) ativadas com as proteínas da superfície celular de S. schenckii (PSCs) em camundongos infectados com S. schenckii strictu sensu. As BMDCs foram estimuladas com PSCs e analisadas quanto à expressão superficial de moléculas co-estimulatórias, bem como à secreção de citocinas pró-inflamatórias. Posteriormente, camundongos sádios foram vacinados com uma ou duas doses de BMDCs para avaliar a sua imunogenicidade e, por último, foi avaliado o efeito das BMDCs em camundongos infectados por S. schenckii. Nossos resultados mostram que as PSCs foram capazes de ativar as BMDCs. A imunização de camundongos saudáveis com BMDCs estimulados por PSCs induziu uma resposta imune com perfil Th17, com aumento da frequência de células IL-17A+ e da liberação de IL-17 e IL-6. Já a vacinação com BMDCs estimuladas com PSCs nos camundongos infectados com S. schenckii levou à redução da carga fúngica no baço desses animais, além do aumento da frequência de células Th IL-17+ e Th IFN+ e uma maior secreção de IL-10, IL-17 e IFN-γ, induzindo uma resposta Th1/Th17 mista. Como um todo, nossos resultados mostram que a vacinação com BMDCs estimuladas com PSCs foi capaz de conferir proteção nos animais vacinados, o que nos leva a crer que é possível, com os ajustes adequados, utilizar DCs como ferramenta terapêutica contra a esporotricose.
Sporotrichosis is a universally distributed mycosis caused by thermodimorphic fungi of the Sporothrix schenckii (S. schenckii) species complex. In Brazil, sporotrichosis is considered endemic and is usually acquired by accidental inoculation of its causative agent through the skin or through zoonotic transmission by infected cats. Clinical forms may vary between cutaneous, lymphocutaneous and systemic, the latter being more commonly observed in immunosuppressed patients. The ineffectiveness of antifungal treatment against this mycosis, especially in immunocompromised patients, has led to the search for more effective and safe therapies. Based on several studies showing the efficient use of dendritic cells as a tool for the development of different fungal vaccines, the aim of this work was to evaluate the protective capacity of bone marrow derived dendritic cells (BMDCs) activated with cell surface proteins of S. schenckii (ScCWP) in mice infected with S. schenckii strictu sensu. The BMDCs were stimulated with PSCs and analyzed for surface expression of costimulatory molecules and the secretion of proinflammatory cytokines. Subsequently, healthy mice were vaccinated with one or two doses of BMDCs to assess their immunogenicity, and finally the effect of BMDCs on S. schenckii infected mice was evaluated. Our results show that the ScCWPs were able to activate BMDCs. Immunization of healthy mice with ScCWPs-stimulated BMDCs induced a Th17 profile immune response, with increased Th IL-17A+ cell frequency and IL-17 and IL-6 release. Vaccination with ScCWPs-stimulated BMDCs in mice infected with S. schenckii led to a reduction in the fungal load in the spleen of these animals, as well as increased frequency of Th IL-17+ and Th IFN+ cells and increased IL-10, IL-17 and IFN-γ secretion, inducing a mixed Th1/Th17 response. Overall, our results show that vaccination with ScCWPs-stimulated BMDCs was able to provide protection in vaccinated animals, leading us to believe that it is possible, with appropriate adjustments, to use DCs as a therapeutic tool against sporotrichosis.
Sporotrichosis is a universally distributed mycosis caused by thermodimorphic fungi of the Sporothrix schenckii (S. schenckii) species complex. In Brazil, sporotrichosis is considered endemic and is usually acquired by accidental inoculation of its causative agent through the skin or through zoonotic transmission by infected cats. Clinical forms may vary between cutaneous, lymphocutaneous and systemic, the latter being more commonly observed in immunosuppressed patients. The ineffectiveness of antifungal treatment against this mycosis, especially in immunocompromised patients, has led to the search for more effective and safe therapies. Based on several studies showing the efficient use of dendritic cells as a tool for the development of different fungal vaccines, the aim of this work was to evaluate the protective capacity of bone marrow derived dendritic cells (BMDCs) activated with cell surface proteins of S. schenckii (ScCWP) in mice infected with S. schenckii strictu sensu. The BMDCs were stimulated with PSCs and analyzed for surface expression of costimulatory molecules and the secretion of proinflammatory cytokines. Subsequently, healthy mice were vaccinated with one or two doses of BMDCs to assess their immunogenicity, and finally the effect of BMDCs on S. schenckii infected mice was evaluated. Our results show that the ScCWPs were able to activate BMDCs. Immunization of healthy mice with ScCWPs-stimulated BMDCs induced a Th17 profile immune response, with increased Th IL-17A+ cell frequency and IL-17 and IL-6 release. Vaccination with ScCWPs-stimulated BMDCs in mice infected with S. schenckii led to a reduction in the fungal load in the spleen of these animals, as well as increased frequency of Th IL-17+ and Th IFN+ cells and increased IL-10, IL-17 and IFN-γ secretion, inducing a mixed Th1/Th17 response. Overall, our results show that vaccination with ScCWPs-stimulated BMDCs was able to provide protection in vaccinated animals, leading us to believe that it is possible, with appropriate adjustments, to use DCs as a therapeutic tool against sporotrichosis.
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Sporothrix schenckii, vacina, células dendríticas, células dendríticas derivadas da medula óssea, esporotricose, Sporothrix schenckii, vaccine, dendritic cells, bone-marrow-derived dendritic cells, sporotrichosis, sporotrichosis