Programa de Pós-graduação em Biologia da Relação Parasito-Hospedeiro
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Navegando Programa de Pós-graduação em Biologia da Relação Parasito-Hospedeiro por Por Orientador "Dias, Fátima Ribeiro"
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Item Avaliação da produção de interleucina 6, fator de necrose tumoral e interleucina 10 em hemoculturas de pacientes com Leishmaniose Cutânea localizada(Universidade Federal de Goiás, 2017-03-10) Costa, Ana Carolina Vieira da; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Cardoso, Ludimila Paula Vaz; Pereira, Ledice Inacia de AraujoAmerican Tegumentary Leishmaniasis (ATL) is caused by protozoa of the genus Leishmania. The parasite is hosted by macrophages in cutaneous or mucosal lesions. Monocytes are precursors of macrophages, but there is little information about monocyte activation during ATL. The present study aimed to evaluate cytokine profile of monocytes from patients with localized cutaneous leishmaniasis (LCL) in whole blood cultures stimulated with toll-like receptor (TLRs) and NOD2 receptor agonists. Peripheral blood from patients with active lesions and no treatment, and from healthy blood donors paired by sex and age, was diluted into culture medium and incubated with lipopolysaccharide (LPS), TLR4 agonist; lipopeptide Pam3Cys (Pam), TLR2 agonist; muramyldipeptide (MDP), NOD2 agonist or Leishmania (V.) braziliensis antigen (Ag). After incubation, supernatants were collected to measure IL-6 and TNF (6 h) and IL-10 (24 h) by ELISA. There was production of IL-6, TNF and IL-10 in cultures from patients and controls after stimulation with all agonists and Ag (p < 0.05, medium vs stimulus, n = 29). No differences were detected between concentrations of TNF and IL-10, but IL-6 was higher produced in non-stimulated cultures from patients than controls (p < 0.05, n = 29) or when the stimulus was MDP (p < 0.05, n = 19). The combination Pam/MDP increased the production of IL-6, TNF and IL-10 in cultures from patients and controls when compared with each stimulus isolated (p < 0.05, n = 13 - 14); similarly, Ag and MDP together induced higher concentrations of IL-6, TNF and IL-10 (IL-6 and IL-10, especially in cultures from patients) in comparison with each stimulus (p < 0.05, n = 11 - 14). There was no correlation between the number, size or time of lesions and levels of IL-6, TNF or IL-10. Patients with more than one cycle of treatment to obtain clinical cure showed higher levels of MDP-induced IL-6 than those cured with one cycle of treatment (p < 0.05, n = 17 cured with one cycle vs 8 cured with more than one cycle). Data suggest that LCL patient monocytes can be activated via TLR2, TLR4 or NOD2 to produce pro-inflammatory (IL-6, TNF) and anti-inflammatory (IL-10) cytokines and IL-6 can be associated with tissue lesion and delay of lesion healing indicating an immunopathogenic role in LCL.Item Investigação dos efeitos do treinamento com ß-glucana no controle da infecção por Leishmania (Viannia) braziliensis em camundongos C57BL/6 transgênicos para interleucina 32ƴ humana(Universidade Federal de Goiás, 2019-03-08) Figueiredo, Ana Marina Barroso de; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; Galdino Junior, Hélio; Gomes, Clayson MouraB-glucan induces trained innate immunity in monocytes/macrophages. Studies suggest that IL-32 is involved in mecanisms important for the control of L. braziliensis and transgenic mouse for the human IL-32 gene (IL-32) is a model for evaluating the functions of this cytokine. Thus, this study aims to evaluate the effect of -glucan training on the control of Leishmania braziliensis infection in IL-32Tg or wild-type (WT) mice. The mice were trained with -glucan and infected with L. braziliensis in the paw (105 or 106 parasites). We evaluated: lesion size, parasite load, histopathological characteristics, cytokines in macerated of the infected paws, lymph node and bone marrow cell cultures. Macrophages were derived from bone marrow precursors from WT or IL-32Tg animals after training in vivo and infected with L. braziliensis to evaluate phagocytosis and leishmanicidal activity. In IL-32Tg mice infected with 105 parasites, when compared to WT animals, -glucan training led to an increase in lesion size on week 3 of infection, associated with an increased inflammatory process and increased production of interferon gamma (IFN); and less parasitic load and less intense inflammatory process after 8 weeks of infection. In bone marrow cell cultures of IL-32Tg animals, 7 days after -glucan injection, there was an increase in IL-1production after stimulation with Leishmania antigens. The phagocytic activity of macrophages from IL-32Tg animals trained with -glucan was higher than that of trained WT animals, but the ability to control infection was similar. Data with inoculum of 105 parasites suggest that IL-32 enhances the effects ofItem Avaliação do papel das citocinas interleucina 32 e interleucina 15 em macrófagos humanos primários infectados com Leishmania (Viannia) braziliensis(Universidade Federal de Goiás, 2016-05-13) Silva, Lucas Luiz de Lima; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; Nagib, Patrícia; Gomes, Rodrigo SaarAmerican Tegumentary Leishmaniasis (ATL) is a disease caused by Leishmania protozoan, belonging to the subgenus Viannia and Leishmania. In Brazil, the most common and prevalent species is L. (V.) braziliensis. In patients with ATL, it was detected the expression of interleukin 32 (IL-32) in skin or mucosal lesions caused by L. Viannia spp. However, the role of IL-32 on ATL is still unclear. It has been shown that IL-15 induces IL-32 and also IL-15 leads to L. infantum control. This study aimed to investigate the effects of IL-32 and IL-15 in the production of cytokines and microbicidal activity of primary human macrophages infected with L. (V.) braziliensis. For this, human peripheral blood monocytes were derived into macrophages and infected with metacyclic forms of L. (V.) braziliensis; evaluation of the infection index (4 h, phagocytosis, 48 h, microbicidal activity) in the absence or presence of rIL-32, rIL-15 or gamma interferon (rIFN) and lipopolysaccharide (LPS); in culture supernatants, IL-32, tumor necrosis factor (TNF) and IL-10 were measured by enzime-linked immunoassay. The addition of rIL-32 to macrophages did not significantly altered phagocytosis of the parasites or microbicidal activity of macrophages. Classical activation of macrophages with rIFN plus LPS decreased the infection index. rIL-32 em high concentration (200 ng/mL) was able to induce TNF in uninfected or infected macrophages, and IL-10 was not induced. Parasites induced lower amounts of intracellular IL-32 as well as rIFN0.1 ng / ml), but there was a synergism between the activation signals provided by the parasites and rIFN (0.1 ng / ml). In the opposite, rIFN in higher concentration (10 ng/mL) induced higher amounts of IL-32, but its activity was partially inhibited by parasites. The rIL-15 was able to induce IL- 32 and TNF in macrophages, but not IL-10 in both non-infected or infected macrophages. The rIL-15 also decreased phagocytosis of parasites by macrophages and increased the microbicidal activity of these cells. The data suggest that IL-15 induces IL-32 and TNF which can contribute to control of the infection. To evaluate the leishmanicidal mechanism pathways induced by IL-15 and IL-32 can help in the development of new therapies for the control of ATL.Item Avaliação do efeito da expressão do gene da interleucina 32 (IL-32) humana em modelo murino de infecção por Leishmania (Leishmania) amazonensis(Universidade Federal de Goiás, 2016-08-24) Silva, Muriel Vilela Teodoro; Gomes, Rodrigo Saar; http://lattes.cnpq.br/8840051460928720; Dias, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403; Dias, Fátima Ribeiro; Santiago, Helton da Costa; Galdino Junior, HélioIL-32 is a proinflammatory cytokine which has different isoforms. IL-32γ isoform is the most powerful and was detected in lesions of patients with cutaneous leishmaniasis. Murine cells respond to IL-32, however mice lack the gene for this cytokine. To understand the role of IL- 32 in Leishmania (L.) amazonensis, we used transgenic mice for human IL-32γ (IL-32γTg). C57BL/6 mice (WT) and C57BL/6 IL-32γTg were infected with L. amazonensis promastigotes in the ear. The lesion development was followed weekly with a digital caliper (measured in mm of injury). After 3, 6 and 9 weeks, the animals were euthanized for tissue parasitism analysis by the limiting dilution technique, in infected ears, draining lymph node and spleen of mice. The draining lymph node cells were incubated (48 h) in the presence or absence of L. amazonensis antigen (Ag) for analysis of cytokines by ELISA. IL-32γTg mice present IL-32 production in spleen, liver, lymph node and ear. IL-32γTg mice have a lower injury than the WT mice during the third week of infection. From the 5th to the 9th week of infection, the two groups had similar lesion development profiles. Interestingly, in the 3rd week of infection, the parasitic load in the lesion of IL-32γTg mice was 100 times greater than that of WT mice. After three weeks, IL-32γTg mice maintained the same parasitic load up to nine weeks. In WT mice, however, the number of parasites increased exponentially during weeks evaluated. The parasite load in the spleen and lymph node was lower in IL-32γTg mice when compared with WT mice. There was no difference in histological sections of the lesions in WT and IL-32γTg mice infected with L. amazonensis. We did not observe differences between WT and IL-32γTg groups on the product -10) by lymph node cells stimulated with Ag, in the 3rd, 6th and 9th week of infection. Our data suggest that IL-32γ favors infection by L. amazonensis in the early stages, allowing the growth of the parasites. However, this cytokine seems to limit the growth and spread of parasites in the later stages of infection. In vitro analyzes show the similar percentage of infected cells and the number of parasites per infected cell in WT macrophages and IL-32γTg after 3 and 48h of infection with L. amazonensis. However, the production of NO by macrophages seems to be lower in IL- 32γTg mouse cells during infection with L. amazonensis. Understanding the mechanisms by which IL-32γ modulates Leishmania amazonensis infection in mice is essential to define the components that control cutaneous leishmaniasis caused by this specie in humans.