Research Project

Project Title:	NETs and terminal complement pathway as potential biomarkers for complement overactivation assessment in ANCA-associated vasculitis
Project Type:	Translational research
Disease group(s):	Immune glomerulopathies
Project Summary:	Clinical, in vitro, and animal model-derived evidence has demonstrated a critical involvement of the alternative complement pathway (aCP) in the pathogenesis of ANCA-associated vasculitis (AAV). In this regard, neutrophil extracellular traps (NETs) have been suggested to be a key element between ANCA-induced neutrophil activation and aCP (1). However, the role of the terminal complement pathway (tCP) is less well studied (2). Methods: A prospective, observational, multicenter study analyzing first episodes and relapses of patients with AAV, with a minimum follow-up of 6 months, is on going . Blood samples are collected at diagnosis (AAV-t1) and at remission (AAV-t2). Control population consistes of age and sex-matched individuals. Complement activation is assessed by analyzing the complement membrane attack complex (C5b-9) deposition on cultured endothelial cells (HMEC-1), by indirect immunofluorescence, after exposing them to activated plasma (patient’s citrated plasma mixed with healthy subjects’ sera pool, 1:1). C5b-9 deposits induced by patient’s activated plasma will be calculated as percentage of labeled area with respect to the total area analyzed. Results from patient and control samples will be expressed as fold increase (mean±SEM) vs. those obtained with the pool of activated plasma from healthy subjects. Plasmatic levels of aCP and tCP soluble factors, such as sFBb and sC5b-9 (respectively), are also measured (mean±SEM). Circulating NETs are indirectly measured by quantifying circulant dsDNA plasmatic concentration (mean±SEM).
Lead principal investigator(s):	Elena Guillen, Barcelona
Co-investigator(s):	Miquel Blasco, Barcelona Luis Quintana, Barcelona
Project Period:	01/2021 - 12/2024
Sponsors:	Regional funding agency

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