Reduced Neutrophil Extracellular Trap Formation During Ischemia Reperfusion Injury in C3 KO Mice: C3 Requirement for NETs Release.

Xiaoting Wu,Danyu You,Liyu Lin,Liyan Yang,Jiong Cui,Jianxin Wan,Yi Chen,Changsheng Xu,Guili Lian

doi:10.3389/fimmu.2022.781273

Abstract

Complement C3 plays a prominent role in inflammatory processes, and its increase exacerbates ischemia reperfusion injury (IRI)-induced acute kidney injury (AKI). Infiltrated neutrophils can be stimulated to form neutrophil extracellular traps (NETs), leading to renal injury. However, the relationship between the increase of C3 and the release of NETs in AKI was not clear. Here we found that IRI in the mouse kidney leads to increased neutrophils infiltration and NET formation. Furthermore, neutrophils depletion by anti-Ly6G IgG (1A8) did not reduce C3 activation but reduced kidney injury and inflammation, indicating a link between neutrophils infiltration and renal tissue damage. Pretreatment with 1A8 suppressed ischemia-induced NET formation, proving that extracellular traps (ETs) in renal tissue were mainly derived from neutrophils. Renal ischemia injury also leads to increased expression of C3. Moreover, C3 KO mice (C3 KO) with IRI exhibited attenuated kidney damage and decreased neutrophils and NETs. In vitro, C3a primed neutrophils to form NETs, reflected by amorphous extracellular DNA structures that colocalized with CitH3 and MPO. These data reveal that C3 deficiency can ameliorate AKI by reducing the infiltration of neutrophils and the formation of NETs. Targeting C3 activation may be a new therapeutic strategy for alleviating the necroinflammation of NETs in AKI.

Highlights

Ischemia–reperfusion injury (IRI) is a major cause of acute kidney injury (AKI), which correlates with high morbidity, mortality, and seriously threatens human health [1]
The formation of extracellular traps (ETs) in the kidney after IRI has been demonstrated to be crucial in the pathogenesis of AKI, the origin of ETs and the relationship between C3 and ETs have not been investigated to date
Our results suggest that following renal IRI, there was an increase in infiltration of neutrophils, formation of neutrophil extracellular traps (NETs), and expression of C3 within the kidney, which correlated with renal injury

Summary

Introduction

Ischemia–reperfusion injury (IRI) is a major cause of acute kidney injury (AKI), which correlates with high morbidity, mortality, and seriously threatens human health [1]. C3 Aggravate Acute Kidney Injury phagocytosis, neutrophils have been confirmed to release neutrophil extracellular traps (NETs). Peptidylarginine deiminase 4 (PAD4) is required for generation of NETs, by promoting histone citrullination and release of decondensed chromatin and granule content [4, 5]. NETs were initially described as a part of the antimicrobial defense and were recently demonstrated to be involved in the pathogenesis of sterile inflammation, such as renal IRI [12]. Neutrophil proteins such as MPO or Ly6g co-expression with citrullinated histone H3 (CitH3) is considered as evidence of NET formation [13]

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Results

Conclusion