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Stimulus-dependent chromatin dynamics, citrullination, calcium signalling and ROS production during NET formation.
de Bont, Cynthia M; Koopman, Werner J H; Boelens, Wilbert C; Pruijn, Ger J M.
Afiliação
  • de Bont CM; Department of Biomolecular Chemistry, Institute of Molecules and Materials (IMM), Radboud University, Nijmegen, the Netherlands.
  • Koopman WJH; Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Nijmegen, the Netherlands.
  • Boelens WC; Department of Biomolecular Chemistry, Institute of Molecules and Materials (IMM), Radboud University, Nijmegen, the Netherlands.
  • Pruijn GJM; Department of Biomolecular Chemistry, Institute of Molecules and Materials (IMM), Radboud University, Nijmegen, the Netherlands. Electronic address: G.Pruijn@ncmls.ru.nl.
Biochim Biophys Acta Mol Cell Res ; 1865(11 Pt A): 1621-1629, 2018 11.
Article em En | MEDLINE | ID: mdl-30327203
Neutrophils can release their chromatin to form neutrophil extracellular traps (NETs), a process known as NETosis. Although NET formation can be induced by various stimuli, recent evidence suggests that these stimuli do so via different mechanisms. Here, we have analysed NET formation induced by lipopolysaccharide (LPS), phorbol 12­myristate 13­acetate (PMA) and the calcium (Ca2+) ionophore A23187. Our results show distinct peroxidase and neutrophil elastase activities in both culture supernatant and NETs. Especially stimulation with A23187 led to pronounced peroxidase and elastase release and yielded high peroxidase activity on the resulting NETs. In contrast to LPS and PMA, A23187 did not induce morphological changes of the nuclei. Histone H3 citrullination was more extensively observed upon induction by A23187 and particularly in LPS- and PMA-induced NETs the detection of citrullinated H3 was dependent on the inhibition of neutrophil proteases, which suggests that NET-associated citrullinated histones are readily cleaved by these proteases. With live cell imaging techniques, differences in the rate of plasma membrane permeabilization were observed, not only for the different inducers, but also among individual neutrophils. LPS and PMA, but not A23187, induced early calcium oscillations and the cytosolic calcium concentrations gradually increased upon LPS and PMA stimulation until the plasma membrane ruptured. The levels of reactive oxygen species rose rapidly after PMA stimulation and much later in neutrophils exposed to LPS and A23187. Taken together, the observed molecular and dynamic differences indicate that NET formation induced by LPS, PMA and A23187 proceeds via different pathways.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cálcio / Espécies Reativas de Oxigênio / Sinalização do Cálcio / Armadilhas Extracelulares / Neutrófilos Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cálcio / Espécies Reativas de Oxigênio / Sinalização do Cálcio / Armadilhas Extracelulares / Neutrófilos Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article