Análise das subpopulações linfocitárias no sedimento urinário de pacientes com nefrite lúpica
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Data
2016-04-19
Autores
Mesquita Junior, Danilo [UNIFESP]
Orientadores
Souza, Alexandre Wagner Silva de [UNIFESP]
Tipo
Tese de doutorado
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Introdução: lúpus eritematoso sistêmico (LES) é uma doença inflamatória crônica de origem autoimune, que se caracteriza pelo envolvimento de diversos órgãos e sistemas. Alterações do sistema imunológico são frequentes no LES, com envolvimento de subpopulações linfocitárias efetoras e regulatórias. A nefrite lúpica (NL) tem impacto significativo na morbidade e mortalidade de pacientes com LES. Marcadores urinários são candidatos para uma avaliação acurada da NL, pois podem representar alteração local imediata do status renal. Objetivos: avaliar a frequência de subpopulações de linfócitos T (LT) CD4+ na urina e no sangue periférico de pacientes com NL e analisar associações com parâmetros clínicos e imunológicos. Métodos: a citometria de fluxo foi utilizada para avaliar a frequência de subpopulações de LT CD4+ na urina e em células mononucleares do sangue periférico (CMSP) de 17 pacientes com NL ativa e em 20 pacientes com nefrite controle (NC), enquanto em 10 controles sadios (CS) foram avaliadas apenas a frequência de subpopulações de LT CD4+ em CMSP. Subpopulações de LT CD4+ no CMSP foram reavaliadas em seis meses de terapia de indução para a NL. Os subtipos de LT em tecido renal foram detectados por imunohistoquímica. Resultados: pacientes com NL apresentam frequência diminuída de LT CD4+ no sangue periférico (SP) quando comparado a NC e CS (p = 0,0001), houve menor frequência de células T regulatórias no SP em relação à NC (p = 0,014). Porém, não houve diferença quanto às subpopulações de LT CD4+ na urina quando comparamos NL e NC. Aos seis meses de tratamento, os pacientes com NL mostraram elevação significativa na frequência de células Th17 no SP (p = 0,008). Formas não proliferativas de NL apresentam frequência aumentada de células Th17 da urina em relação às formas proliferativas (p = 0,041). A frequência de células que expressam Tbet, ROR-? e GATA3 no glomérulo de pacientes com NL é significativamente maior do que de células FOXP3+. Não houve associação entre expressão glomerular desses marcadores e parâmetros clínicos ou laboratoriais na NL. Todos os pacientes com NL apresentaram expressão de CD3 e Tbet no interstício renal, enquanto a expressão de FOXP3 e ROR-? só foi observada em 30% e 10% dos pacientes, respectivamente. Houve correlação negativa entre células Th1 no SP e células Th1 urinárias (Rho=-0,531; p = 0,028), entre células Th1 no SP e as células Tbet+ no interstício renal (Rho=-0,782; p = 0,004). Conclusões: células Th1 tem papel importante na patogênese da NL, migrando para o tecido renal e para urina. Células Th17 se associam a curso menos grave da NL e são induzidas pela terapia. Diferenças na distribuição de células CD4+ entre NL e NC foram observadas apenas no SP e não na urina. Esses achados auxiliam a compreensão do processo fisiopatológico da NL.
Introduction: systemic erythematosus lupus (SLE) is a chronic inflammatory disease characterized by the involvement of multiple organs and systems. Immunology alterations are frequently observed in SLE, especially involving effector and regulatory lymphocytes subsets. Lupus nephritis (LN) has a significant impact on morbidity and mortality of SLE patients. Urinary markers are a potential tool for accurate assessment of LN, since they may reflect immediate local alterations in renal status. Objectives: to evaluate the frequency of urinary and peripheral blood (PB) CD4+ T cells subsets in patients with LN and analyze associations with clinical and immunological parameters. Methods: flow cytometry was used to assess the frequency of urinary and PB CD4+ T cells subsets in 17 SLE patients with active LN and in 20 patients with other forms of glomerulonephritis, named as nephritis control (NC). In 10 healthy controls (HC), the frequency of CD4+ T cells subtypes was evaluated only in PB. CD4+ T cells subsets in PB were reevaluated at six months of induction therapy in LN patients. T cell subsets were detected in renal tissue by immunohistochemistry. Results: the frequency of CD4+ T cells was lower in LN patients compared with NC and HC (p = 0,0001), whereas regulatory T cells in PB of LN patients was lower than in NC patients (p = 0.014). However, no difference could be found when we compared urinary CD4+ T cells subsets between LN and NC. At six months of treatment, LN patients showed an increased frequency of Th17 cells in PB (p = 0.008). Nonproliferative forms of LN show increased frequency of Th17 cells in urine compared with proliferative forms (p = 0.041). The expression of Tbet, GATA3 and RORγ in glomeruli is significantly higher than FOXP3. No association was found between the expression of T cells subsets markers in glomeruli and clinical or laboratory parameters in LN. In renal interstice, CD3+ and Tbet+ cells are present in all LN patients who underwent renal biopsy while FOXP3 and RORγ were observed in 30% and 10% of LN patients, respectively. An inverse correlation was found between Th1 in PB and urinary Th1 cells (Rho=0.531; p = 0.028) and Th1 cells from PB are inversely correlated with renal tissue Tbet expression (Rho=0.782; p = 0.004). Conclusion: Th1 cells have an important role in LN pathogenesis, migrating from PB to renal tissue and urine. Th17 cells are associated with less serious LN course and are increased in PB by therapy. Differences in the CD4+ cells distribution comparing LN and NC were observed only in PB but not in urine. These findings provides a better understanding of the pathophysiological process of LN.
Introduction: systemic erythematosus lupus (SLE) is a chronic inflammatory disease characterized by the involvement of multiple organs and systems. Immunology alterations are frequently observed in SLE, especially involving effector and regulatory lymphocytes subsets. Lupus nephritis (LN) has a significant impact on morbidity and mortality of SLE patients. Urinary markers are a potential tool for accurate assessment of LN, since they may reflect immediate local alterations in renal status. Objectives: to evaluate the frequency of urinary and peripheral blood (PB) CD4+ T cells subsets in patients with LN and analyze associations with clinical and immunological parameters. Methods: flow cytometry was used to assess the frequency of urinary and PB CD4+ T cells subsets in 17 SLE patients with active LN and in 20 patients with other forms of glomerulonephritis, named as nephritis control (NC). In 10 healthy controls (HC), the frequency of CD4+ T cells subtypes was evaluated only in PB. CD4+ T cells subsets in PB were reevaluated at six months of induction therapy in LN patients. T cell subsets were detected in renal tissue by immunohistochemistry. Results: the frequency of CD4+ T cells was lower in LN patients compared with NC and HC (p = 0,0001), whereas regulatory T cells in PB of LN patients was lower than in NC patients (p = 0.014). However, no difference could be found when we compared urinary CD4+ T cells subsets between LN and NC. At six months of treatment, LN patients showed an increased frequency of Th17 cells in PB (p = 0.008). Nonproliferative forms of LN show increased frequency of Th17 cells in urine compared with proliferative forms (p = 0.041). The expression of Tbet, GATA3 and RORγ in glomeruli is significantly higher than FOXP3. No association was found between the expression of T cells subsets markers in glomeruli and clinical or laboratory parameters in LN. In renal interstice, CD3+ and Tbet+ cells are present in all LN patients who underwent renal biopsy while FOXP3 and RORγ were observed in 30% and 10% of LN patients, respectively. An inverse correlation was found between Th1 in PB and urinary Th1 cells (Rho=0.531; p = 0.028) and Th1 cells from PB are inversely correlated with renal tissue Tbet expression (Rho=0.782; p = 0.004). Conclusion: Th1 cells have an important role in LN pathogenesis, migrating from PB to renal tissue and urine. Th17 cells are associated with less serious LN course and are increased in PB by therapy. Differences in the CD4+ cells distribution comparing LN and NC were observed only in PB but not in urine. These findings provides a better understanding of the pathophysiological process of LN.
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Citação
MESQUITA JUNIOR, Danilo. Análise das subpopulações linfocitárias no sedimento urinário de pacientes com nefrite lúpica. 2016. 106 f. Tese (Doutorado em Ciências da Saúde Aplicadas à Reumatologia) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, 2016.