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2015 ; 181
(3
): 518-27
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Neutrophil extracellular traps can activate alternative complement pathways
#MMPMID25963026
Wang H
; Wang C
; Zhao MH
; Chen M
Clin Exp Immunol
2015[Sep]; 181
(3
): 518-27
PMID25963026
show ga
The interaction between neutrophils and activation of alternative complement
pathway plays a pivotal role in the pathogenesis of anti-neutrophil cytoplasmic
antibody (ANCA)-associated vasculitis (AAV). ANCAs activate primed neutrophils to
release neutrophil extracellular traps (NETs), which have recently gathered
increasing attention in the development of AAV. The relationship between NETs and
alternative complement pathway has not been elucidated. The current study aimed
to investigate the relationship between NETs and alternative complement pathway.
Detection of components of alternative complement pathway on NETs in vitro was
assessed by immunostain and confocal microscopy. Complement deposition on NETs
were detected after incubation with magnesium salt ethyleneglycol tetraacetic
acid (Mg-EGTA)-treated human serum. After incubation of serum with supernatants
enriched in ANCA-induced NETs, levels of complement components in supernatants
were measured by enzyme-linked immunosorbent assay (ELISA). Complement factor B
(Bb) and properdin deposited on NETs in vitro. The deposition of C3b and C5b-9 on
NETs incubated with heat-inactivated normal human serum (Hi-NHS) or EGTA-treated
Hi-NHS (Mg-EGTA-Hi-NHS) were significantly less than that on NETs incubated with
NHS or EGTA-treated NHS (Mg-EGTA-NHS). NETs induced by ANCA could activate the
alternative complement cascade in the serum. In the presence of EGTA, C3a, C5a
and SC5b-9 concentration decreased from 800·42?±?244·81 ng/ml, 7·68?±?1·50 ng/ml,
382·15?±?159·75 ng/ml in the supernatants enriched in ANCA induced NETs to
479·07?±?156·2 ng/ml, 4·86?±?1·26 ng/ml, 212·65?±?44·40 ng/ml in the supernatants
of DNase I-degraded NETs (P?
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