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Influência do soro de doentes com Lúpus Eritematoso Sistémico na activação in vitro de monócitos e células dendríticas
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Authors
Abstract(s)
Introdução: O Lúpus Eritematoso Sistémico (LES) é uma doença auto-imune
caracterizada por uma desregulação do sistema imune, que leva a uma produção
aumentada de anticorpos e formação de imuno-complexos, resultando na
produção de citocinas pró-inflamatórias e dano celular de diferentes órgãos.
Objectivos: Este trabalho teve como objectivos a avaliação do efeito
estimulatório provocado pelo soro de doentes com LES em monócitos e células
dendríticas (DCs) do sangue periférico de controlos saudáveis e a quantificação
de TNF-α no soro de doentes com LES e de um grupo controlo.
Material e Métodos: Foram incluídos neste estudo 21 doentes com LES,
agrupados de acordo com o índice de actividade da doença, em doentes com
doença activa (LESA) - 11 e inactiva (LESI) - 11, bem como 10 indivíduos
saudáveis que constituíram o grupo controlo. A expressão de TNF-α e IL-6 em
monócitos, mDCs e DCs CD16+CD14-/low obtidas do sangue periférico de
indivíduos saudáveis foi avaliada por citometria de fluxo, após estimulação in vitro
com LPS e na presença de soro de doentes com LES e de um grupo controlo. O
estudo da expressão génica de TNF-α foi realizado após separação das
diferentes populações celulares em estudo por qRT-PCR. A concentração sérica
de TNF-α foi determinada em 64 doentes com LES e 27 controlos, quantificada
através da MILLIPLEX MAP Human TH17 Magnetic Bead Panel Premixed.
Resultados: Observou-se que o soro de doentes com LES, particularmente
aqueles com um maior título de ac. anti-dsDNA, provocava um aumento da
expressão de TNF-α, nos monócitos e principalmente nas DCs CD16+CD14-/low.
Também se observou um aumento da expressão de mRNA de TNF-α nesta
última população celular, bem como um aumento da concentração sérica de
TNF-α nos doentes com LES.
Conclusão: Este estudo enfatiza o papel do ambiente inflamatório a nível
periférico nos doentes com LES e a sua importância na activação das células
circulantes.
Introduction: Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by dysregulation of the immune system which leads to an increased production of antibodies and formation of immune complexes, resulting in production of pro-inflammatory cytokines and cell damage in different organs. Aims: The aim of this study was evaluate the stimulatory effect induced by sera from SLE patients in monocytes and dendritic cells (DCs) from healthy individuals and quantify of soluble TNF- α in sera from SLE patients and in a control group. Material and Methods: In this study were enrolled 21 SLE patients, classified according to the index of disease activity in patients with active disease (ASLE) - 11, and patients with inactive disease (ISLE) – 11, as well as 10 healthy individuals as a group control. The expression of TNF-α and IL-6 in monocytes, mDCs and CD16+CD14-/low DCs obtained from peripheral blood of healthy individuals was analysed, after in vitro stimulation with LPS in the presence of SLE patients serum, by flow cytometry. The TNF-α gene expression was performed after cell sorting by qRT-PCR. Soluble TNF-α was determinated in 64 SLE patients and 27 healthy control by MILLIPLEX MAP Human TH17 Magnetic Bead Panel Premixed. Results: It was observed a higher TNF-α expression in monocytes but particulary in CD16+CD14-/low DCs, when stimulated in the presence of SLE patients serum, particularly in those with high levels of anti-dsDNA. We also observed an increase mRNA expression of TNF-α in CD16+CD14-/low DCs, as well as a higher soluble TNF-α in the sera from SLE patients. Conclusion: This study emphasizes the role of the inflammatory environment at the peripheral level in SLE patients and its importance in the activation of circulating cells.
Introduction: Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by dysregulation of the immune system which leads to an increased production of antibodies and formation of immune complexes, resulting in production of pro-inflammatory cytokines and cell damage in different organs. Aims: The aim of this study was evaluate the stimulatory effect induced by sera from SLE patients in monocytes and dendritic cells (DCs) from healthy individuals and quantify of soluble TNF- α in sera from SLE patients and in a control group. Material and Methods: In this study were enrolled 21 SLE patients, classified according to the index of disease activity in patients with active disease (ASLE) - 11, and patients with inactive disease (ISLE) – 11, as well as 10 healthy individuals as a group control. The expression of TNF-α and IL-6 in monocytes, mDCs and CD16+CD14-/low DCs obtained from peripheral blood of healthy individuals was analysed, after in vitro stimulation with LPS in the presence of SLE patients serum, by flow cytometry. The TNF-α gene expression was performed after cell sorting by qRT-PCR. Soluble TNF-α was determinated in 64 SLE patients and 27 healthy control by MILLIPLEX MAP Human TH17 Magnetic Bead Panel Premixed. Results: It was observed a higher TNF-α expression in monocytes but particulary in CD16+CD14-/low DCs, when stimulated in the presence of SLE patients serum, particularly in those with high levels of anti-dsDNA. We also observed an increase mRNA expression of TNF-α in CD16+CD14-/low DCs, as well as a higher soluble TNF-α in the sera from SLE patients. Conclusion: This study emphasizes the role of the inflammatory environment at the peripheral level in SLE patients and its importance in the activation of circulating cells.
Description
Keywords
Lupus Eritematoso Sistémico Soro Monócitos Células dendríticas TNF-α IL-6 Systemic Lupus Erythematosus Serum Monocytes Dendritic cells