Browsing by Author "Antonelli, Lis Ribeiro do Valle"

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    Innovations in diagnosis and post-therapeutic monitoring of Chagas disease : simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi.
    (2014) Alessio, Glaucia Diniz; Côrtes, Denise Fonseca; Assis, Girley Francisco Machado de; Sales Júnior, Policarpo Ademar; Ferro, Eloisa Amália Vieira; Antonelli, Lis Ribeiro do Valle; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Lana, Marta de
    This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with falsepositive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeuticmonitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% fromsamples of the treated cured (TC) patientswere negativewith AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease.
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    Recrutamento do receptor de adenosina A2B e ativação da via de AMPc- PI3K-ERK1/2 inibem a resposta de células dendríticas infectadas por Leishmania amazonensis.
    (2016) Figueiredo, Amanda Braga de; Afonso, Luís Carlos Crocco; Gonçalves, Teresa Maria Fonseca de Oliveira; Cunha, Rodrigo Pinto dos Santos Antunes da; Brandão, Rogélio Lopes; Oliveira, Milton Adriano Pelli de; Faria, Ana Maria Caetano de; Antonelli, Lis Ribeiro do Valle
    A infecção por Leishmania pode resultar num amplo espectro de manifestações clínicas e a infecção por Leishmania amazonensis é associada a uma deficiência na resposta de linfócitos T específicos para o parasito. Células dendríticas direcionam a diferenciação de linfócitos Th1 que contribuem para o controle da infecção por Leishmania. Em um trabalho anterior, nós mostramos que a infecção por L. amazonensis, mas não por L. braziliensis ou L. major, prejudica a resposta de células dendríticas por um mecanismo dependente do receptor de adenosina A2B. Neste trabalho, nós avaliamos a expressão de receptores de adenosina e os eventos intracelulares ativados a partir do receptor A2B em células dendríticas infectadas, bem como o papel desses eventos na infecção de camundongos por L. amazonensis. Inicialmente, células dendríticas derivadas de células de medula óssea de camundongos C57BL/6J foram infectadas por promastigotas metacíclicas de L. amazonensis, L. braziliensis ou L. major. Imagens obtidas por microscopia de fluorescência mostraram que a infecção por promastigotas metacíclicas de L. amazonensis estimula a redistribuição do receptor A2B para a superfície de células dendríticas infectadas, sem alterar a quantidade total do receptor, avaliada por western blotting, nem a expressão dos receptores de adenosina A1, A2A e A3. A infecção por L. braziliensis, L. major ou promastigotas procíclicas de L. amazonensis não alteram a expressão do receptor A2B em células infectadas. Nossos resultados mostraram, ainda, que a infecção por L. amazonensis estimula a produção de AMPc e a fosforilação de ERK1/2 por mecanismos dependentes do receptor A2B. Além disso, L. amazonensis prejudica a expressão de CD40 e a produção de IL-12p70 por células dendríticas, efeitos independentes de IL-10 e revertidos pela inibição de adenilato ciclase, de PI3K e da fosforilação de ERK1/2. Por fim, camundongos infectados na orelha por L. amazonensis na presença de inibidores do receptor A2B ou das proteínas PI3K ou ERK1/2 desenvolvem lesões menores se comparados a animais controles, mas apenas o bloqueio do receptor A2B é capaz de diminuir o parasitismo tecidual. Concluindo, nós propomos um novo mecanismo utilizado por L. amazonensis (redistribuição do receptor A2B AMPc PI3K ERK1/2) para inibir a ativação de células dendríticas, que parece ser importante para a deficiência na resposta imune observada na infecção por essa espécie de parasito.
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    Systemic Immunological changes in patients with distinct clinical outcomes during Mycobacterium tuberculosis infection.
    (2017) Papini, Tatiane Figueiredo Morais; Reis, Jordana Grazziela Alves Coelho dos; Wendling, Ana Paula Barbosa; Antonelli, Lis Ribeiro do Valle; Wowk, Pryscilla Fanini; Bonato, Vânia Luiza Deperon; Augusto, Valéria Maria; Santos, Silvana Maria Elói; Martins Filho, Olindo Assis; Carneiro, Cláudia Martins; Carvalho, Andréa Teixeira de
    Background: The lung lesions in an individual infected with tuberculosis (TB) are surprisingly variable and independent of each other. However, there is no circulating biomarker yet able to segregate patients according to the extent of lung lesions. Materials and methods: In this study, the phenotypic and functional profile of leukocytes of patients with active pulmonary tuberculosis (TB) and controls (CO) were fully scrutinized by immunophenotyping assays and in vitro short-term whole blood culture. The TB group was subdivided according to the extent of lung lesions as unilateral (UNI) and bilateral (BI). Results: The results show that TB group display an altered leukocyte profile in the peripheral blood with significant lower counts of NK-cells, CD3+CD56+CD16+/− NKT-cells, CD4+T-cells, CD19+B-cells when compared to CO. Increased CD4+T-cells and CD8+T-cell activation was observed by the upregulation of activation markers (HLA-DR) as well as of chemokine receptors (CCR2, CCR3, and CXCR4). In addition, TB group presented a significant decrease proportion of CD14LowCD16+ monocytes despite the increase in HLA-DR expression. Regarding the severity of the disease, in the BI group a reduction in frequency of CD19+CD5+ B-cells and expression of HLA-DR in CD14LowCD16+ monocytes was observed. Furthermore, the extent of lung lesions influences the production of molecules as observed by significantly larger production of IL-4 by neutrophils, total T-cells, CD4+T-cells, CD8+T-cells and CD19+B-cells in UNI as compared to BI. By contrast, in BI group the frequency of high producers of both IL-17+CD4+T-cells and IL-17+CD8+T-cells were significantly increased than UNI, suggesting the deleterious role of these subsets during active pulmonary Mtb infection. Conclusion: The immunophenotypic characterization of unilateral and bilateral active TB performed in the present study indicates that the extent of lung lesion could be associated with a fine-tuning between immunological responses during untreated Mtb infection.