Estudo da resposta imune de camundongos BALB/c com a proteína recombinante A2 de Leishmania chagasi

• Ano de defesa: 2015
• Autor(a) principal: Jusi, Márcia Mariza Gomes [UNESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Leishmania; Camundongo como animal de laboratorio; Resposta imune; Proteínas recombinantes; Genes; Immune response
• Link de acesso: http://hdl.handle.net/11449/126394

The causative agent of the disease in Brazil is Leishmania chagasi. It is transmitted by the bite of the sand fly Lutzomyia longipalpis, which acquires the parasite to realize the hematophagism in infected animals. This work aimed to study the immune response of BALB / c mice immunized with the recombinant protein produced from A2-L gene of L. chagasi, isolated sample of a dog the Veterinary Hospital of FCAV-UNESP, Jaboticabal and to evaluate the ability of this recombinant protein to induce immunoprotection in animals after challenge with the parasite. Forty mice were divided into four groups: G1, 10 animals inoculated with L. chagasi promastigotes; G2, 10 animals inoculated with the recombinant protein A2-L; G3, 10 animals immunized with the recombinant protein A2-L and challenged with promastigotes of L. chagasi; G4, 10 animals inoculated with saline (negative control). Characteristics evaluated humoral immune response were antibodies of the IgG class and IgG1 and IgG2a subclasses, the indirect ELISA. As for the cellular immune response, we evaluated the production of CD4+ and CD8+ by flow cytometry technique and evaluated the immunoblots of CD4+, CD8+, iNOS, macrophages, MHC I, MHC II and B lymphocytes, by the technique of immunohistochemistry. The production of cytokines (IL-2, IFN-γ, TNF-α, IL-4 and IL-10) by RT-PCR in real time and, the parasite load of the spleens of mice infected with the parasite by qPCR. The G3 had a pattern of humoral response Th1 = Th2, while the G1, the pattern was Th1< Th2. The G3 group also showed higher production of CD8 cells and cells expressing MHC I, MHC II and iNOS. The group G3 excels in the production of IL-2, IFN-γ and TNF-α, mRNA the spleen cells. The parasite load was significantly lower in the group G3, less than half of that found in G1. We therefore conclude that the G3 group produced a pattern of humoral and cellular responses, which according to literature suggests be effective ...