Biological activity and chemical composition of the ethanolic extracts of Miconia ferruginata DC. / Atividade biológica e composição química dos extratos etanólicos de Miconia ferruginata DC.

Ethnopharmacological relevance: Miconia ferruginata DC. is a native plant from Brazilian Cerrado biome known as “pixirica” or “babatenã”, widely used in traditional medicine as an anti-inflammatory and antibiotic agent. Aim of the study: This study aimed to perform a preliminary analysis of the chemical profile and screening of biological activities of the ethanolic extracts of the leaves/flowers (EELF) and of the stem (EES) of this species. Materials and methods: The techniques Ultra-Fast Liquid Chromatography with diode-array detection (UFLC-DAD) and High-Performance Liquid Chromatography with diode-array detection and mass spectrometry (HPLC-DAD-MS) performed chemical analysis. Biological activities evaluated for the antibacterial, antitripanosamatides and antitumor effect through in vitro assays, by MTT and resazurin.Results: Although the extracts showed a negligible result for antibacterial and antitripanosamatides effect, this species showed a high cytotoxicity against tumor cells (p< 0.001) of 4T1, A549 and? MDA-MB-231, associated with low cell toxicity against fibroblasts. High concentration of phenolic compounds detected in the extracts, especially flavonoids derivates from quercetin, catechins and phenolic acids. Conclusion: These phenolic compounds have a high biological potential and may be responsible for the observed cytoxicity, together the data suggest the M. ferruginata has a great potential for being one promising candidate for further studies against cancer

Pediatric Patients With Steroid-Sensitive Nephrotic Syndrome Have Higher Expression of T Regulatory Lymphocytes in Comparison to Steroid-Resistant Disease

Background and Aim: Idiopathic nephrotic syndrome (INS) is classified according to the response to drug therapy in steroid-sensitive (SS), steroid-dependent (SD), and steroid-resistant (SR) categories. Previous studies showed changes in inflammatory activity of subpopulations of lymphocytes in INS. This study aimed to compare SS and SR patients in regard to subpopulations of leukocytes, profile of regulatory lymphocytes, and migratory activity of lymphocyte subpopulations. Results obtained in INS patients were also compared to age and sex-matched healthy controls.Methods: This is a cross-sectional study including SS patients (n = 30), SR patients (n = 14), and controls (n = 10). Peripheral blood samples were withdrawn for ex-vivo leukocyte flow cytometry analysis.Results: Percentage of B-lymphocytes and natural killer (NK) cells were significantly reduced in SR patients when compared to controls, while the percentage of NKT cells were decreased in SS patients in comparison to controls. Percentages of CD4+ expressing FoxP3 and CTLA4 were significantly higher in SS patients in comparison to SR patients and controls. The expression of integrin CD18 on the surface of T lymphocytes (CD3+) was reduced in SS patients if compared to controls.Conclusion: This study found that SS INS patients have higher levels of regulatory T-lymphocytes and lower expression of adhesion molecules than SR patients

New insights into Trypanosoma cruzi genetic diversity, and its influence on parasite biology and clinical outcomes

Chagas disease, caused by Trypanosoma cruzi, remains a serious public health problem worldwide. The parasite was subdivided into six distinct genetic groups, called “discrete typing units” (DTUs), from TcI to TcVI. Several studies have indicated that the heterogeneity of T. cruzi species directly affects the diversity of clinical manifestations of Chagas disease, control, diagnosis performance, and susceptibility to treatment. Thus, this review aims to describe how T. cruzi genetic diversity influences the biology of the parasite and/or clinical parameters in humans. Regarding the geographic dispersion of T. cruzi, evident differences were observed in the distribution of DTUs in distinct areas. For example, TcII is the main DTU detected in Brazilian patients from the central and southeastern regions, where there are also registers of TcVI as a secondary T. cruzi DTU. An important aspect observed in previous studies is that the genetic variability of T. cruzi can impact parasite infectivity, reproduction, and differentiation in the vectors. It has been proposed that T. cruzi DTU influences the host immune response and affects disease progression. Genetic aspects of the parasite play an important role in determining which host tissues will be infected, thus heavily influencing Chagas disease’s pathogenesis. Several teams have investigated the correlation between T. cruzi DTU and the reactivation of Chagas disease. In agreement with these data, it is reasonable to suppose that the immunological condition of the patient, whether or not associated with the reactivation of the T. cruzi infection and the parasite strain, may have an important role in the pathogenesis of Chagas disease. In this context, understanding the genetics of T. cruzi and its biological and clinical implications will provide new knowledge that may contribute to additional strategies in the diagnosis and clinical outcome follow-up of patients with Chagas disease, in addition to the reactivation of immunocompromised patients infected with T. cruzi

Análise do perfil fenotípico e funcional de leucócitos do sangue periférico de crianças com síndrome nefrótica idiopática

Exportado OPUSMade available in DSpace on 2019-08-14T17:53:57Z (GMT). No. of bitstreams: 1 tese_final_para_cpg.pdf: 2596384 bytes, checksum: 83f3daefbcebdd732609c98f223e0c53 (MD5) Previous issue date: 31A síndrome nefrótica idiopática (SNI) se caracteriza por proteinúria maciça, hipoproteinemia e edema. Considerando as características imunopatológicas da SNI, o presente estudo teve como objetivo analisar o perfil fenotípico e funcional de leucócitos do sangue periférico de crianças com SNI. Foram estudados 44 pacientes pediátricos com SNI, provenientes da Unidade de Nefrologia Pediátrica do Hospital das Clínicas da Universidade Federal de Minas Gerais e 10 crianças e adolescentes saudáveis utilizados como grupo controle (CON). Os pacientes foram subdivididos de acordo com os valores da proteinúria em urina de 24 horas no momento da coleta em grupo com proteinúria elevada (PE200mg/24 horas) e proteinúria reduzida (PR<200mg/24 horas) e de acordo com a resposta à corticoterapia em córtico-sensíveis (SNCS) e córtico-resistentes/dependentes (SNCR). Os pacientes e controles foram submetidos à coleta de sangue para análise ex-vivo do perfil fenotípico de leucócitos circulantes por citometria de fluxo. Foi também analisada a produção in vitro de citocinas pró e anti-inflamatórias em populações específicas de linfócitos do sangue periférico. Foram comparados os dados obtidos em pacientes e no grupo controle e de acordo com a intensidade da proteinúria. Além disso, foram avaliados o comportamento migratório e de ativação celular e o percentual de células T com perfil regulador em linfócitos de pacientes com síndrome nefrótica e do grupo controle. Tais parâmetros foram comparados nos subgrupos de SNCS e SNCR, bem como conforme a proteinúria. Os resultados mostraram aumento no percentual de linfócitos TCD4+TNF-+ e redução no percentual de linfócitos TCD8+IFN-+ nos pacientes com SNI, independente dos níveis de proteinúria. Nos pacientes com PE, houve aumento no percentual de linfócitos TCD4+ marcados para as citocinas pró-inflamatórias IFN-, TNF- e IL-17 quando comparado ao grupo controle. Somente os pacientes com PE apresentaram aumento no percentual de linfócitos TCD8+TNF-+ em comparação ao grupo controle. Os achados sugerem que os pacientes com proteinúria persistente apresentam resposta pró-inflamatória mais acentuada e mediada por linfócitos T, principalmente pelos linfócitos T CD4+, mesmo sob uso de medicação anti-inflamatória. A comparação dos pacientes subdivididos conforme a resposta à corticoterapia mostrou redução nos percentuais de linfócitos B e células NK nos pacientes com SNCR quando comparado ao grupo CON. Já o grupo com SNCS apresentou apenas redução no percentual de células NKT quando comparado ao CON. Em linfócitos T CD4+, a expressão de FoxP3 nos linfócitos TCD4+ foi significativamente maior nos pacientes do grupo SNCS quando comparado aos grupos CON e SNCR. O percentual de células CD4+CTLA4+ foi maior no grupo SNCS em relação ao SNCR. Foi observada redução na expressão da integrina CD18 em linfócitos TCD3+ e TCD8+ no grupo SNCS quando comparado ao CON. A análise dos receptores de quimiocinas, no entanto, revelou um aumento no percentual de linfócitos marcados para CCR2 e CXCR4 tanto no grupo SNCS quanto no SNCR quando comparados ao grupo controle. O aumento no percentual de células marcadas para o receptor CCR5 foi observado apenas no grupo SNCR comparado ao CON. A redução da expressão de CD18 na superfície de linfócitos TCD3 e TCD8, bem como as evidências de um perfil regulador em linfócitos TCD4 presente apenas em pacientes com SNCS sugerem que o controle da resposta inflamatória nesse grupo de pacientes possa estar relacionado a alterações nesses parâmetros. O aumento na expressão de receptores de quimiocinas CCR2 e CCR4 em todos os pacientes com SNI pode ser explicado pela natureza inflamatória da doença. Por outro lado, apenas o grupo com SNCR apresentou aumento na expressão de CCR5, indicando atividade infamatória maior ainda nesse grupo de pacientes. Em conjunto, nossos resultados sugerem que a capacidade de ativar mecanismos reguladores da resposta imune esteja associada à sensibilidade aos corticosteroides.Idiopathic nephrotic syndrome (INS) is a disease characterized by massive proteinuria, hypoproteinemia and edema. Considering the immunopathological characteristics of INS, this study aimed to analyze the phenotypic and functional profile of peripheral blood leukocytes of pediatric patients with INS. We evaluated 44 pediatric patients with INS recruited at the Pediatric Nephrology Unit of the Clinics Hospital from Federal University of Minas Gerais and 10 health children and adolescent as control group (CON). Pacients were subdivided according to the levels of proteinuria at the time of blood collection in high proteinuria (HP 200mg/24 hours) and low proteinuria (LP<200mg/24 hours); and also based on the response to corticosteroids in steroid sensitive (SS) and steroid resistant/dependent (SR). Patients and controls were submitted to blood collection for ex-vivo analysis of the phenotype profile of circulating leukocytes by flow cytometry. In vitro production of pro- and anti-inflammatory cytokines in specific lymphocytes populations from peripheral blood was also analyzed. Data were compared between patients and controls and in patients according the level of proteinuria. In addition, the migratory and celular activation behavior and the percentage of T-cells with regulatory profile were evaluated in lymphocytes of INS patients and controls. These parameters were compared in the subgroups of SS and SR patients, as well according to the proteinuria. Our results showed an increase of the percentage of CD4+TNF-+ lymphocytes and a decrease of the percentage of CD8+IFN-+ lymphocytes in patients with INS, regardless the levels of proteinuria. In patients with HP, there was increased percentage of CD4+ lymphocytes stained to pro-inflammatory cytokines, IFN-, TNF- and IL-17, as compared to control group. Only patients with HP exhibited an increase in the percentage of CD8+TNF-+ lymphocytes as compared to control group. These results suggest that patients with persistent proteinuria have more pronounced pro-inflammatory response, mediated primarily by TCD4+ lymphocytes, even under anti-inflammatory medication. The comparison of patients divided according to steroid response showed reduced percentages of B lymphocytes and NK cells in SR group as compared to control group. SS patients had lower percentage of NKT cells than controls. In CD4+ lymphocytes, the expression of the regulatory marker FoxP3 was significantly higher in SS patients than controls and SR group. The percentage of CD4+CTLA4+ cells was higher in SS patients than in SR group. Reduced expression of the migration marker CD18 was observed in CD3+ and CD8+ T lymphocytes of the SS group as compared to controls. The analysis of chemokine receptors revealed increased percentage of lymphocytes stained for CCR2 and CXCR4 both in SS and SR groups as compared to the control group. The increased percentage of cells stained for CCR5 receptor was observed only in SR group as compared to controls. Reduced expression of CD18 on the surface of TCD3 and TCD8 lymphocytes, as well the higher percentage of TCD4 cells with regulatory profile only in SS patients suggest that the control of inflammatory response in this group of patients might be related to changes in these parameters. Higher expression of chemokine receptors, CCR2 and CCR4, in INS patients may be explained by the inflammatory nature of the disease. On the other hand, only SR group showed an increased expression of CCR5, indicating higher inflammatory activity in this subgroup of patients. Taken together, our results suggest that the ability to activate regulatory mechanisms of immune response would be associated with steroid sensitivity

Análise de receptores de quimiocinas na superfície de leucócitos circulantes de indivíduos infectados pelo Mycobacterium leprae: resultados preliminares

Neste estudo, a expressão de receptores de quimiocinas na superfície dos leucócitos circulantes foi feita pela citometria de fluxo. Houve aumento da porcentagem de linfócitos CCR2+CD4+ no sangue periférico dos pacientes com hanseníase. Este resultado preliminar sugeriu alteração do perfil dos receptores de quimiocinas desses pacientes

Increased Migratory and Activation Cell Markers of Peripheral Blood Lymphocytes in an Experimental Model of Nephrotic Syndrome

The present study aimed to evaluate the expression of CD80 and CD18 in subpopulations of peripheral blood leukocytes and oxidative kidney damage in rats with nephrotic syndrome (NS) induced by doxorubicin (Dox) in comparison to control animals at different time points. Male adult Wistar rats were submitted to 24-hour urine and blood collection for biochemical and immunological analysis at 7, 14, 21, and 28 days after Dox injection. After euthanasia, the kidneys were removed for histological analysis and the evaluation of oxidative stress. The phenotypic characterization of leukocytes was performed using flow cytometry. Dox-injected animals exhibited increased CD18 expression in cytotoxic T lymphocytes, NK cells, and monocytes and high CD80 expression in monocytes. Kidney oxidative damage was positively correlated with CD80 expression in monocytes and serum levels of creatinine. These results suggest that phagocytic and cytotoxic cells are preferentially recruited to the tissue injury site, which may contribute to kidney dysfunction in this animal model of NS. The blockade of integrin and costimulatory molecules may provide new therapeutic opportunities for NS

Trypan blue exclusion assay by flow cytometry

Submitted by Nuzia Santos ([email protected]) on 2015-01-28T15:27:26Z No. of bitstreams: 1 2014_003.pdf: 1135145 bytes, checksum: dc2e947d746266f16712e0890b86299c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-01-28T15:27:34Z (GMT) No. of bitstreams: 1 2014_003.pdf: 1135145 bytes, checksum: dc2e947d746266f16712e0890b86299c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-01-28T15:33:09Z (GMT) No. of bitstreams: 1 2014_003.pdf: 1135145 bytes, checksum: dc2e947d746266f16712e0890b86299c (MD5)Made available in DSpace on 2015-01-28T15:33:09Z (GMT). No. of bitstreams: 1 2014_003.pdf: 1135145 bytes, checksum: dc2e947d746266f16712e0890b86299c (MD5) Previous issue date: 2014Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Diamantina, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Diamantina, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Fisiologia e Farmacologia. Belo Horizonte, MG, Brasil.Fundação Osvaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Biomarcadores de Diagnóstico e Monitoração Belo Horizonte, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Diamantina, MG, Brasil.Universidade Federal dos Vales do Jequitinhonha e Mucuri. Departamento de Farmácia. Laboratório de Imunologia. Diamantina, MG, Brasil.Dye exclusion tests are used to determine the number of live and dead cells. These assays are based on the principle that intact plasma membranes in live cells exclude specific dyes, whereas dead cells do not. Although widely used, the trypan blue (TB) exclusion assay has limitations. The dye can be incorporated by live cells after a short exposure time, and personal reliability, related to the expertise of the analyst, can affect the results. We propose an alternative assay for evaluating cell viability that combines the TB exclusion test and the high sensitivity of the flow cytometry technique. Previous studies have demonstrated the ability of TB to emit fluorescence when complexed with proteins. According to our results, TB/bovine serum albumin and TB/cytoplasmic protein complexes emit fluorescence at 660 nm, which is detectable by flow cytometry using a 650-nm low-pass band filter. TB at 0.002% (w/v) was defined as the optimum concentration for distinguishing unstained living cells from fluorescent dead cells, and fluorescence emission was stable for 30 min after cell treatment. Although previous studies have shown that TB promotes green fluorescence quenching, TB at 0.002% did not interfere with green fluorescence in human live T-cells stained with anti-CD3/fluorescein isothiocyanate (FITC) monoclonal antibody. We observed a high correlation between the percentage of propidium iodide+CD3/FITC+ and TB+CD3/FITC+ cells, as well as similar double-stained cell profiles in flow cytometry dot-plot graphs. Taken together, the results indicate that a TB exclusion assay by flow cytometry can be employed as an alternative tool for quick and reliable cell viability analysis

Dimethyl Sulfoxide (DMSO) Decreases Cell Proliferation and TNF-α, IFN-γ, and IL-2 Cytokines Production in Cultures of Peripheral Blood Lymphocytes

Dimethylsulfoxide (DMSO) is an amphipathic molecule composed of a polar domain characterized by the sulfinyl and two nonpolar methyl groups, for this reason it is able to solubilize polar and nonpolar substances and transpose hydrophobic barriers. DMSO is widely used to solubilize drugs of therapeutic applications and studies indicated that 10% v/v concentration did not modify culture viability when used to treat human peripheral blood mononuclear cells (PBMC). However, some DMSO concentrations could influence lymphocyte activation and present anti-inflammatory effects. Therefore, the objective of this study was to evaluate the effect of DMSO on lymphocyte activation parameters. Cell viability analysis, proliferation, and cytokine production were performed on PBMC from six healthy subjects by flow cytometry. The results indicated that 2.5% v/v DMSO concentrations did not modify lymphocytes viability. DMSO at 1% and 2% v/v concentrations reduced the relative proliferation index of lymphocytes and at 5% and 10% v/v concentrations reduced the percentage of total lymphocytes, cluster of differentiation 4+ (CD4+) T lymphocytes and CD8+ T lymphocytes interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) producers. Thus, it was concluded that DMSO has an in vitro anti-inflammatory effect by reducing lymphocyte activation demonstrated with proliferation reduction and the decrease of cytokine production

T‐lymphocyte‐expressing inflammatory cytokines underlie persistence of proteinuria in children with idiopathic nephrotic syndrome

Objective: There is evidence of an important role of immune system changes in the triggering and maintenance of idiopathic nephrotic syndrome (INS). The aim of this study was to investigate the expression of cytokines in lymphocyte populations of patients with INS in comparison to healthy individuals, according to proteinuria. Methods: This cross‐sectional study included 44 patients with INS and eight healthy children, matched for age and sex (controls). Patients were subdivided according to proteinuria: persistent proteinuria or partial remission (PP ≥ 300 mg/24 h, n = 17) and low proteinuria or complete remission (LP < 300 mg/24 h, n = 27). Ex vivo analysis of peripheral blood leukocytes by flow cytometry was performed using surface markers for T‐lymphocytes, TCD4, TCD8, natural killer (NK) cells, NKT, and B‐lymphocytes. Frequencies of intracellular cytokines were analyzed in these cells. Results: The frequencies of B‐lymphocytes, NK cells, and NKT cells were lower in INS than in controls, whereas INS patients had a higher frequency of CD4+tumor necrosis factor (TNF)‐α+ cells than controls. Cytotoxic‐T‐lymphocytes expressing IFN‐γ were lower in INS than in controls. Patients with PP showed higher frequencies of CD4‐T‐lymphocytes expressing IFN‐γ and TNF‐α than controls. CD8‐lymphocytes expressing TNF‐α were increased in PP group when compared with LP and controls, while CD8+interferon (IFN)‐γ+ cells were lower than in LP and in controls. Conclusion: Regardless the level of proteinuria, INS patients had increased expression of TNF‐α in CD4‐lymphocytes and reduced expression of IFN‐γ in CD8‐lymphocytes. Persistence of proteinuria was associated with higher levels of inflammatory markers. Resumo: Objetivo: Há comprovação do importante papel das alterações no sistema imunológico no desencadeamento e na manutenção da síndrome nefrótica idiopática (SNI). O objetivo deste estudo foi investigar a expressão das citocinas em populações de linfócitos de pacientes com SNI em comparação a indivíduos saudáveis e de acordo com a proteinúria. Métodos: Este estudo transversal incluiu 44 pacientes com SNI e oito crianças saudáveis, pareados por idade e sexo (controles). Os pacientes foram subdivididos de acordo com a proteinúria: proteinúria persistente ou remissão parcial (PP ≥ 300 mg/24 h, n = 17) e proteinúria baixa ou remissão completa (PB < 300 mg/24 h, n = 27). A análise ex vivo de leucócitos no sangue periférico por citometria de fluxo foi feita utilizando marcadores de superfície para linfócitos T, TCD4, TCD8, células natural killer (NK), linfócitos NKT e B. As frequências das citocinas intracelulares foram analisadas nessas células. Resultados: A frequência dos linfócitos B, células NK e células NKT foi menor em pacientes com SNI do que nos controles, ao passo que os pacientes com SNI apresentaram maior frequência de células CD4+fator de necrose tumoral (TNF)‐α+ do que nos controles. Os linfócitos T citotóxicos que expressam interferon (IFN)‐γ foram menores nos pacientes com SNI do que nos controles. Os pacientes com PP mostraram maiores frequências de linfócitos T CD4 que expressam IFN‐γ e TNF‐α que os controles. Os linfócitos CD8 que expressam TNF‐α apresentaram aumento no grupo com PP, em comparação aos com PB e os controles, apesar de as células CD8+IFN‐γ+ serem mais baixas nos pacientes com PB e nos controles. Conclusão: Com relação ao nível de proteinúria, os pacientes com SNI apresentaram aumento na expressão de TNF‐α nos linfócitos CD4 e expressão reduzida de IFN‐γ nos linfócitos CD8. A persistência da proteinúria foi associada a maiores níveis de marcadores inflamatórios. Keywords: Idiopathic nephrotic syndrome, Cytokines, Inflammation, Proteinuria, Immune response, Palavras‐chave: Síndrome nefrótica idiopática, Citocinas, Inflamação, Proteinúria, Resposta imun

T-lymphocyte-expressing inflammatory cytokines underlie persistence of proteinuria in children with idiopathic nephrotic syndrome

Objective: There is evidence of an important role of immune system changes in the triggering and maintenance of idiopathic nephrotic syndrome (INS). The aim of this study was to investigate the expression of cytokines in lymphocyte populations of patients with INS in comparison to healthy individuals, according to proteinuria. Methods: This cross-sectional study included 44 patients with INS and eight healthy children, matched for age and sex (controls). Patients were subdivided according to proteinuria: persistent proteinuria or partial remission (PP ≥ 300 mg/24 h, n = 17) and low proteinuria or complete remission (LP < 300 mg/24 h, n = 27). Ex vivo analysis of peripheral blood leukocytes by flow cytometry was performed using surface markers for T-lymphocytes, TCD4, TCD8, natural killer (NK) cells, NKT, and B-lymphocytes. Frequencies of intracellular cytokines were analyzed in these cells. Results: The frequencies of B-lymphocytes, NK cells, and NKT cells were lower in INS than in controls, whereas INS patients had a higher frequency of CD4+tumor necrosis factor (TNF)-α+ cells than controls. Cytotoxic-T-lymphocytes expressing IFN-γ were lower in INS than in controls. Patients with PP showed higher frequencies of CD4-T-lymphocytes expressing IFN-γ and TNF-α than controls. CD8-lymphocytes expressing TNF-α were increased in PP group when compared with LP and controls, while CD8+interferon (IFN)-γ+ cells were lower than in LP and in controls. Conclusion: Regardless the level of proteinuria, INS patients had increased expression of TNF-α in CD4-lymphocytes and reduced expression of IFN-γ in CD8-lymphocytes. Persistence of proteinuria was associated with higher levels of inflammatory markers. Resumo: Objetivo: Há comprovação do importante papel das alterações no sistema imunológico no desencadeamento e manutenção da síndrome nefrótica idiopática (SNI). O objetivo deste estudo foi investigar a expressão das citocinas em populações de linfócitos de pacientes com SNI em comparação a indivíduos saudáveis e de acordo com a proteinúria. Métodos: Este estudo transversal incluiu 44 pacientes com SNI e oito crianças saudáveis, pareados por idade e sexo (controles). Os pacientes foram subdivididos de acordo com a proteinúria: proteinúria persistente ou remissão parcial (PP ≥ 300 mg/24 h, n = 17) e proteinúria baixa ou remissão completa (PB < 300 mg/24 h, n = 27). A análise ex vivo de leucócitos no sangue periférico por citometria de fluxo foi feita utilizando marcadores de superfície para linfócitos T, TCD4, TCD8, células natural killer (NK), linfócitos NKT e B. As frequências das citocinas intracelulares foram analisadas nessas células. Resultados: A frequência dos linfócitos B, células NK e células NKT foi menor em pacientes com SNI do que nos controles, ao passo que os pacientes com SNI apresentaram maior frequência de células CD4+fator de necrose tumoral (TNF)-α+ do que nos controles. Os linfócitos T citotóxicos que expressam interferon (IFN)-γ foram menores nos pacientes com SNI do que nos controles. Os pacientes com PP mostraram maiores frequências de linfócitos T CD4 que expressam IFN-γ e TNF-α que os controles. Os linfócitos CD8 que expressam TNF-α apresentaram aumento no grupo com PP, em comparação aos com PB e os controles, apesar de as células CD8+IFN-γ+ serem mais baixas nos pacientes com PB e nos controles. Conclusão: Com relação ao nível de proteinúria, os pacientes com SNI apresentaram aumento na expressão de TNF-α nos linfócitos CD4 e expressão reduzida de IFN-γ nos linfócitos CD8. A persistência da proteinúria foi associada a maiores níveis de marcadores inflamatórios. Keywords: Idiopathic nephrotic syndrome, Cytokines, Inflammation, Proteinuria, Immune response, Palavras-chave: Síndrome nefrótica idiopática, Citocinas, Inflamação, Proteinúria, Resposta imun