Immunoregulatory mechanisms in Chagas disease: modulation of apoptosis in T-cell mediated immune responses

Submitted by Nuzia Santos ([email protected]) on 2017-07-17T17:53:45Z No. of bitstreams: 1 Chaves_AnaThereza_Immunoregulatory mechanisms_IRR_2016.pdf: 12736177 bytes, checksum: 7182dae7e3675c77254aa3dd4157a0a9 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2017-07-17T18:03:09Z (GMT) No. of bitstreams: 1 Chaves_AnaThereza_Immunoregulatory mechanisms_IRR_2016.pdf: 12736177 bytes, checksum: 7182dae7e3675c77254aa3dd4157a0a9 (MD5)Made available in DSpace on 2017-07-17T18:03:09Z (GMT). No. of bitstreams: 1 Chaves_AnaThereza_Immunoregulatory mechanisms_IRR_2016.pdf: 12736177 bytes, checksum: 7182dae7e3675c77254aa3dd4157a0a9 (MD5) Previous issue date: 2016Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, Brazil/Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Laboratório de Biologia das Interações Celulares. Belo Horizonte, MG, Brazil/Universidade Federal de Minas Gerais. Faculdade de Medicina Programa de Pós graduação em Medicina Tropical e Infectologia. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Biomarcadores de Diagnóstico e Monitoração. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Laboratório de Biologia das Interações Celulares. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Imunologia e Genômica de Parasitos. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Biofísica. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Imunologia e Genômica de Parasitos. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Faculdade de Medicina Programa de Pós graduação em Medicina Tropical e Infectologia. Belo Horizonte, MG, Brazil.Laboratório de Imunologia Celular e Molecular, Centro de Pesquisas René Rachou, Fiocruz, Belo Horizonte, Brazil/Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, Brazil/Universidade Federal de Ouro Preto. Ouro Preto, MG, Brazil.BACKGROUND: Chronic Chagas disease presents different clinical manifestations ranging from asymptomatic (namely indeterminate) to severe cardiac and/or digestive. Previous results have shown that the immune response plays an important role, although no all mechanisms are understood. Immunoregulatory mechanisms such as apoptosis are important for the control of Chagas disease, possibly affecting the morbidity in chronic clinical forms. Apoptosis has been suggested to be an important mechanism of cellular response during T. cruzi infection. We aimed to further understand the putative role of apoptosis in Chagas disease and its relation to the clinical forms of the disease. METHODS: Apoptosis of lymphocytes, under antigenic stimuli (soluble T. cruzi antigens - TcAg) where compared to that of non-stimulated cells. Apoptosis was evaluated using the expression of annexin and caspase 3(+) by T cells and the percentage of cells positive evaluated by flow cytometry. In addition activation and T cell markers were used for the identification of TCD4(+) and TCD8(+) subpopulations. The presence of intracellular and plasma cytokines were also evaluated. Analysis of the activation status of the peripheral blood cells showed that patients with Chagas disease presented higher levels of activation determined by the expression of activation markers, after TcAg stimulation. PCR array were used to evaluate the contribution of this mechanism in specific cell populations from patients with different clinical forms of human Chagas disease. RESULTS: Our results showed a reduced proliferative response associated a high expression of T CD4(+)CD62L(-) cells in CARD patients when compared with IND group and NI individuals. We also observed that both groups of patients presented a significant increase of CD4(+) and CD8(+) T cell subsets in undergoing apoptosis after in vitro stimulation with T. cruzi antigens. In CARD patients, both CD4(+) and CD8(+) T cells expressing TNF-α were highly susceptible to undergo apoptosis after in vitro stimulation. Interestingly, the in vitro TcAg stimulation increased considerably the expression of cell death TNF/TNFR superfamily and Caspase family receptors genes in CARD patients. CONCLUSIONS: Taken together, our results suggest that apoptosis may be an important mechanism for the control of morbidity in T. cruzi infection by modulating the expression of apoptosis genes, the cytokine environment and/or killing of effector cells

Achados histopatológicos em úteros e ovários de cadelas submetidas à castração eletiva pelas técnicas de ovariectomia ou ovariohisterectomia

Objetivou-se avaliar os achados histopatológicos dos ovários e úteros de cadelas submetidas à ovariectomia (OVE) ou ovariohisterectomia (OVH) visando identificar afecções ovarianas e/ou uterinas não diagnosticadas durante exame clínico, laboratorial e ultrassonográfico pré-operatório ou macroscopicamente no transcirúrgico. Foram selecionadas 20 cadelas, clinicamente saudáveis e não prenhes provenientes da casuística do Hospital Veterinário da Universidade Federal Rural de Pernambuco, que foram divididas em Grupo OVH (10 animais) e Grupo OVE (10 animais). Foram coletados os ovários e úteros obtidos durante o procedimento de OVH, e ovários e dois fragmentos de 0,5 cm do corno uterino direito e esquerdo coletados durante a OVE, sendo estes encaminhados para avaliação histopatológica. Os resultados demonstraram a presença de hiperplasia endometrial cística em 80% dos animais submetidos à OVH e em 20% dos animais do grupo OVE. No grupo OVE, foi identificado a presença de hiperplasia adenomatosa de rete ovarii em uma paciente (10%) e adenoma de rete ovarii em outra (10%), alterações estas, consideradas raras na medicina veterinária. Conclui-se que o exame histopatológico é fundamental no diagnóstico de alterações ovarianas e/ou uterinas, não observadas em exame clínico, ultrassonográfico, laboratorial e macroscópico de cadelas submetidas à castração eletiva pelas técnicas de OVH e OVE, e que essas alterações devem direcionar na decisão da técnica cirúrgica a ser utilizada

Avaliação da frequência de genótipos polimórficos em genes codificadores para FOXP3 e sua associação com as diferentes formas clínicas da Doença de Chagas

Exportado OPUSMade available in DSpace on 2019-08-12T03:33:24Z (GMT). No. of bitstreams: 1 disserta__o_karine_silvestre_ferreira.pdf: 1211979 bytes, checksum: c4fb82aba21c6bd046c4fd382c42b72a (MD5) Previous issue date: 9A identificação da subpopulação de células T CD4+CD25highFOXP3+ e do seu papel como células T reguladoras (TREG) têm sido objeto de diversos estudos, devido ao papel crítico dessas células na manutenção da auto-tolerância, bem como na defesa contra infecções. Os mecanismos de supressão mediados por células T reguladoras CD4+CD25high FOXP3+ são ainda pouco compreendidos. De fato, estudos in vitro sobre a caracterização das células TREG em animais e humanos favorecem a hipótese de que o mecanismo de ação destas células pode depender do contato célula-célula e/ou de citocinas. A expressão de FOXP3 tem sido descrita como um importante fator para o desenvolvimento e atividade funcional dessas células. Embora a função dessa população celular ainda não esteja clara na doença de Chagas, estudos anteriores já demonstraram que a frequência de células TCD4+CD25high FOXP3+ é maior em pacientes portadores da forma clínica indeterminada (IND). Assim, o objetivo principal deste estudo é investigar a associação entre polimorfismos do gene FOXP3 e o desenvolvimento de formas clínicas graves da doença de Chagas. Neste trabalho, o DNA extraído a partir do sangue periférico de pacientes com as formas clínicas indeterminada e cardíaca (CARD) da doença de Chagas foi utilizado para analisar a presença de polimorfismos funcionais do gene FOXP3. Ensaios de PCR em tempo real foram conduzidos, utilizando iniciadores dirigidos para o SNPS -3279 C/T e -3499 G/T, localizados no intron-1 do gene FOXP3. Em indivíduos do sexo feminino as frequências genotípicas e alélicas foram associadas com diferentes formas clínicas da doença de Chagas. O estudo do polimorfismo -3499 G / T mostrou que o genótipo heterozigoto (GT) está associado com a forma clínica IND (p= 0,04). Outras análises mostraram a associação da ocorrência do alelo polimórfico (T + -3499 G / T) com a forma clínica indeterminada (p = 0,016, OR =0,295), sugerindo um papel protetor para o polimorfismo avaliado. Este perfil é associado com a elevada expressão do FOXP3 em células TREG. Os nossos resultados sugerem que polimorfismos funcionais no gene FOXP3 em células TREG podem ter um papel importante na infecção pelo T. cruzi, provavelmente através do controle da resposta imune e consequentemente controlando a morbidade.The identification of the CD4+CD25high FOXP3+ subset and of its role as regulatory T cells (TREG) has been the object of intense studies due to the putative critical role of these cells in maintaining self-tolerance, as well as in defense against infections. The suppressive mechanisms mediated by CD4+CD25high FOXP3+ T regulatory cells are not yet understood. In fact, in vitro studies on the characterization of TREG cells in mice and humans favor the hypothesis that the mechanisms of action these cells depends on cell-cell contact and/or on cytokines. FOXP3 expression by these cells has been described as an important factor for their development and functional activities. Although there are no evidences for a clear function of this cell population in Chagas disease, we have previously demonstrated that the frequency of CD4+CD25high FOXP3+ T cells is augmented in patients with the indeterminate clinical form (IND). Thus, the objective of this study is to investigate the association between FOXP3 gene polymorphisms and the development of severe clinical forms of Chagas disease. In this study, DNA extracted from peripheral blood of patients with the IND and cardiac (CARD) clinical forms of Chagas disease were used to analyze the presence of functional polymorphisms of the FOXP3 gene. Real-time PCR using primers directed to the SNPS -3279 C/T and -3499 G/T, located in the intron-1 of the FOXP3 gene were conducted. In female individuals genotypic and allelic frequencies were associated with different clinical forms of Chagas disease. The study of -3499 G/T polymorphisms showed that heterozygosity of this genotype (GT) was associated with IND patients (p=0,04). Other analyses showed an association between the occurrence of polymorphic allele (T+ -3499 G/T) and the IND clinical form (p=0,016 , OR=0,295), suggesting a protective role for the evaluated polymorphism. This profile is associated with high expression of FOXP3 in TREG. Our results suggest that functional polymorphisms in the FOXP3 gene in TREG cells may have an important role in T. cruzi infection, probably by controlling the exacerbated immune response and consequently controlling morbidity

Marcadores de ativação da doença e estado de hipercoagulabilidade no lúpus eritematoso sistêmico

Exportado OPUSMade available in DSpace on 2019-08-12T22:30:56Z (GMT). No. of bitstreams: 1 fafar_tese_karine_2017.pdf: 2128230 bytes, checksum: 1ec11dea1d5c7d51f9d674f96830ddf6 (MD5) Previous issue date: 22O lúpus eritematoso sistêmico (LES) é uma doença autoimune e inflamatória caracterizada pela produção de diversos autoanticorpos e depósitos teciduais de complexos antígeno-anticorpo circulantes. A descoberta e validação de métodos para determinação de produtos de ativação do complemento ligados a células do sangue (CBCAPs= Cell-bound complement activation products), tais como C4d ligados a reticulócitos (R-C4d) e a plaquetas (P-C4d), como potenciais biomarcadores do estado de atividade e do potencial trombótico do LES, respectivamente, podem ser importantes ferramentas para o melhor entendimento da fisiopatologia e do manejo clínico dessa doença. Outro fator importante na patogênese do LES, são os eventos consequentes a anormalidades do sistema hemostático que necessitam ser melhor entendidos. Dessa forma, o objetivo principal desse estudo foi avaliar se, no LES, os níveis de CB-CAPs podem ser preditores de ativação da doença (R-C4d) e de predisposição trombótica (PC4d). Além dos níveis de CB-CAPs, investigou-se o potencial trombótico por meio da determinação dos níveis plasmáticos de importantes componentes do sistema hemostático, tais como dímero D (DDi), trombomodulina (TM), proteína S (PS) e proteína ligadora de C4b (C4BP), buscando-se uma possível associação entre estes parâmetros e os outros investigados. Para alcançar esse objetivo, foram incluídas um total de 60 pacientes com LES, sob tratamento, selecionadas no Serviço de Reumatologia da Santa Casa de Belo Horizonte, sendo 30 pacientes classificadas com doença em baixa atividade (SLEDAI-2K 4) e 30 com a doença em moderada/alta atividade (SLEDAI-2K > 4), enquanto simultaneamente foram selecionadas 30 mulheres sem LES (controles), pareadas por idade e do mesmo nível sócio econômico. As amostras de sangue coletadas das participantes do estudo foram utilizadas para a determinação dos níveis de CB-CAPs por meio da citometria de fluxo e para a obtenção de plasma para a realização dos ensaios previstos neste estudo. Foi observado aumento dos níveis de R-C4d, P-C4d, TM e DDi em pacientes com LES com moderada/alta atividade, cujos níveis correlacionaram com o aumento do índice SLEDAI-2K. Os níveis de PS e C4BP foram semelhantes entre os pacientes com LES moderada/alta atividade, baixa atividade e controles. A análise conjunta dos dados permitiu concluir que níveis elevados de R-C4d e P-C4d correlacionaram com o índice SLEDAI-2K e, portanto, com piora clínica em pacientes com LES. Além do mais, a combinação de RC4d e P-C4d permitiu o estabelecimento de uma proposta de cutoff como indicativo de doença em atividade. Finalmente, foi possível ainda confirmar a relação entre doença em atividade e estado de hipercoagulabilidade, provavelmente associado à dano endotelial e inflamação.Systemic lupus erythematosus (SLE) is an autoimmune and inflammatory disease characterized by the production of various autoantibodies and tissue deposits of circulating antigen-antibody complexes. The discovery and validation of methods for the determination of cell bound complement activation products (CB-CAPs), such as C4d bound to reticulocytes (R-C4d) and to platelets (P-C4d), as potential biomarkers of the activity state and the thrombotic potential of SLE, respectively, may be important tools for a better understanding of the pathophysiology and clinical management of SLE patients. Another important factor in the pathogenesis of SLE are the events resulting from hemostatic system abnormalities that need to be better understood. Thus, the main objective of this study was to evaluate whether the levels of CB-CAPs in SLE patients can be predictors of disease activity (R-C4d) and thrombotic predisposition (P-C4d). In addition to the CB-CAPs levels, the thrombotic potential was investigated by determining the plasma levels of important components of the hemostatic system such as D-dimer (DDi), thrombomodulin (TM), protein S (PS) and C4b binding protein (C4BP), seeking a possible association between these parameters and the others also investigated. To achieve this goal, a total of 60 patients with SLE, under treatment, selected at the Hospital Santa Casa Rheumatology Service in Belo Horizonte was included, with 30 LES patients classified as having low-disease activity (SLEDAI-2K 4) and 30 patients with moderate/high activity (SLEDAI-2K> 4), while 30 women without SLE (controls), matched by age and socioeconomic status were simultaneously selected. Blood samples collected from the study participants were used to determine the levels of CB-CAPs by means of flow cytometry and to obtain plasma for carrying out the other tests included in this study. Compared to both controls and SLE patients with low activity disease, increased levels of R-C4d, P-C4d, TM and DDi were observed in patients with SLE with moderate/high activity disease, whose levels correlated with the increase in the SLEDAI-2K index. PS and C4BP levels were similar among all three groups. Data analysis showed that high levels of R-C4d and P-C4d correlated with the SLEDAI-2K index and, therefore, with clinical worsening in SLE patients. Moreover, the combination of R-C4d and P-C4d allowed the establishment of a cutoff proposal as indicative of disease activity. Finally, it was possible to confirm the relationship between disease activity and hypercoagulability, probably associated with endothelial damage and inflammation

Plasma Cytokine Expression Is Associated with Cardiac Morbidity in Chagas Disease

Submitted by Nuzia Santos ([email protected]) on 2015-02-06T13:25:14Z No. of bitstreams: 1 2014_048.pdf: 2077033 bytes, checksum: 5c9786393949bf065beeeb0aad05f1bd (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-06T13:25:21Z (GMT) No. of bitstreams: 1 2014_048.pdf: 2077033 bytes, checksum: 5c9786393949bf065beeeb0aad05f1bd (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-02-06T14:27:57Z (GMT) No. of bitstreams: 1 2014_048.pdf: 2077033 bytes, checksum: 5c9786393949bf065beeeb0aad05f1bd (MD5)Made available in DSpace on 2015-02-06T14:27:57Z (GMT). No. of bitstreams: 1 2014_048.pdf: 2077033 bytes, checksum: 5c9786393949bf065beeeb0aad05f1bd (MD5) Previous issue date: 2014Universidade Federal de Minas Gerais. Faculdade de Medicina. Infectologia e Medicina Tropical. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Faculdade de Medicina. Infectologia e Medicina Tropical. Belo Horizonte, MG, Brazil/Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Morfologia. Belo Horizonte, Minas Gerais, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Morfologia. Belo Horizonte, Minas Gerais, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Morfologia. Belo Horizonte, Minas Gerais, BrazilUniversidade Federal de Minas Gerais. Faculdade de Medicina. Infectologia e Medicina Tropical. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Morfologia. Belo Horizonte, Minas Gerais, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, MG, Brazil/Instituto Nacional de Ciencia e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, BrazilUniversidade Federal de Minas Gerais. Faculdade de Medicina. Infectologia e Medicina Tropical. Belo Horizonte, MG, BrazilThe expression of immune response appears to be associated with morbidity in Chagas disease. However, the studies in this field have usually employed small samples of patients and statistical analyses that do not consider the wide dispersion of cytokine production observed in these patients. The aim of this study was to evaluate the plasma cytokine levels in welldefined clinical polar groups of chagasic patients divided into categories that better reflect the wide cytokine profile and its relationship with morbidity. Patients infected with Trypanosoma cruzi (T. cruzi) were grouped as indeterminate (IND) and cardiac (CARD) forms ranging from 23 to 69 years of age (mean of 45.6611.25). The IND group included 82 individuals, ranging from 24 to 66 years of age (mean of 39.6610.3). The CARD group included 94 patients ranging from 23 to 69 years of age (mean of 48612.52) presenting dilated cardiomyopathy. None of the patients have undergone chemotherapeutic treatment, nor had been previously treated for T. cruzi infection. Healthy non-chagasic individuals, ranging from 29 to 55 years of age (mean of 42.668.8) were included as a control group (NI). IND patients have a higher intensity of interleukin 10 (IL-10) expression when compared with individuals in the other groups. By contrast, inflammatory cytokine expression, such as interferon gamma (IFN-c), tumor necrosis factor alpha (TNF-a), interleukin 6 (IL-6), and interleukin 1 beta (IL-1b), proved to be the highest in the CARD group. Correlation analysis showed that higher IL-10 expression was associated with better cardiac function, as determined by left ventricular ejection fraction and left ventricular diastolic diameter values. Altogether, these findings reinforce the concept that a fine balance between regulatory and inflammatory cytokines represents a key element in the establishment of distinct forms of chronic Chagas disease

Matrix Metalloproteinases 2 and 9 Are Differentially Expressed in Patients with Indeterminate and Cardiac Clinical Forms of Chagas Disease

Made available in DSpace on 2015-09-28T13:02:37Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) luciana_garzoni_etal_IOC_2013.pdf: 1928957 bytes, checksum: 7cfd5f51da07c835cf156cca694e6005 (MD5) Previous issue date: 2013Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Ultra-Estrutura Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Ultra-Estrutura Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Morfologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Faculdade de Medicina. Programa de Pós-Graduação em Ciências da Saúde, Infectologia e Medicina Tropical. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Faculdade de Medicina. Programa de Pós-Graduação em Ciências da Saúde, Infectologia e Medicina Tropical. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil / Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais INCT-DT. Belo Horizonte, MG, Brasil.Dilated chronic cardiomyopathy (DCC) from Chagas disease is associated with myocardial remodeling and interstitial fibrosis, resulting in extracellular matrix (ECM) changes. In this study, we characterized for the first time the serum matrix metalloproteinase 2 (MMP-2) and MMP-9 levels, as well as their main cell sources in peripheral blood mononuclear cells from patients presenting with the indeterminate (IND) or cardiac (CARD) clinical form of Chagas disease. Our results showed that serum levels of MMP-9 are associated with the severity of Chagas disease. The analysis of MMP production by T lymphocytes showed that CD8 T cells are the main mononuclear leukocyte source of both MMP-2 and MMP-9 molecules. Using a new 3-dimensional model of fibrosis, we observed that sera from patients with Chagas disease induced an increase in the extracellular matrix components in cardiac spheroids. Furthermore, MMP-2 and MMP-9 showed different correlations with matrix proteins and inflammatory cytokines in patients with Chagas disease. Our results suggest that MMP-2 and MMP-9 show distinct activities in Chagas disease pathogenesis. While MMP-9 seems to be involved in the inflammation and cardiac remodeling of Chagas disease, MMP-2 does not correlate with inflammatory molecules

Correlation analysis between plasma IL-10 levels and echocardiographic variable markers of cardiac morbidity.

<p>Correlation analysis between plasma IL-10 levels and cardiac function variables (LVEF and LVDD) in the IND (n = 82, first column) and CARD (n = 94, second column) groups. Correlation analysis was performed using the Spearman correlation coefficient, and results were considered significant when <i>P</i><0.05. Significant differences (<i>P</i>-value) are indicated in each graph together with the <i>r</i> value.</p

Analyses of plasma cytokine levels and their association with cardiac morbidity expressed by the clinical classification.

<p>The analysis of plasma levels was performed as described in the material and methods section. The groups evaluated were: NI (n = 24, white box), IND (n = 82, light gray box), and CARD (n = 94, dark gray box). The results were expressed by mean intensity of fluorescence (MIF). (A) Plasma IL-10 levels in NI, IND, and CARD groups and their association with cardiac morbidity. (B) Plasma IFN-γ levels in NI, IND, and CARD groups and their association with cardiac morbidity. (C) Plasma TNF-α levels in NI, IND, and CARD groups and their association with cardiac morbidity. (D) Plasma IL-6 levels in NI, IND, and CARD groups and their association with cardiac morbidity. E) Plasma IL-1β levels in NI, IND, and CARD groups and their association with cardiac morbidity. Significant differences (<i>P</i>-value<0.05) in the charts are identified by connecting lines and the symbol (*) for comparisons between the groups.</p

Establishing the concept of low, medium and high cytokine producers.

<p>A) Representative scatter plot graph of plasma IL-10 used to establish the cut-off to define low, medium, and high cytokine producers. B) Representative scatter plot graph of plasma IFN-γ used to establish the cut-off edge to define low, medium, and high cytokine producers. C) Representative scatter plot graph of plasma TNF-α used to establish the cut-off to define low, medium, and high cytokine producers. D) Representative scatter plot graph of plasma IL-6 used to establish the cut-off to define low, medium, and high cytokine producers. E) Representative scatter plot graph of plasma IL-1β used to establish the cut-off to define low, medium, and high cytokine producers. Low cytokine producers were defined by values of lower than the first tertile. Medium cytokine producers were defined by values equal to or lower than the second tertile, while high cytokine producers were defined by values higher than or equal to the second tertile. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087082#s3" target="_blank">Results</a> were considered significant with a <i>P</i>-value<0.05.</p

Correlation analysis between plasma IFN-γ, TNF-α, IL-6 levels, and echocardiographic variable markers of cardiac morbidity.

<p>A) Correlation analysis between plasma IFN-γ and cardiac function variables in the IND (n = 82, first column) and CARD (n = 94, second column) groups. B) Correlation analysis between plasma TNF-α and cardiac function variables in the IND (n = 82, first column) and CARD (n = 94, second column) groups. C) Correlation analysis between plasma IL-6 and cardiac function variables in the IND (n = 82, first column) and CARD (n = 94, second column) groups. Correlation analysis was performed using the Spearman correlation coefficient, and results were considered significant when <i>P</i><0.05. Significant differences (<i>P-</i>value) are indicated in each graph together with the <i>r</i> value.</p