Early neutrophil activation and NETs release in the pristane-induced lupus mice model

PLoS One. 2025 Jan 3;20(1):e0306943. doi: 10.1371/journal.pone.0306943. eCollection 2025.

Abstract

Background: NETosis is recognized as an important source of autoantigens. Therefore, we hypothesized whether the pristane-induced lupus mice model shows early activation of neutrophils, the presence of low-density granulocytes (LDGs), and neutrophil extracellular traps (NETs) release, which could contribute to the development of a lupus phenotype.

Methods: Twelve female wild-type Balb/c mice were intraperitoneally injected with pristane (n = 6; pristane group) or saline (n = 6; control group). Five days after the injection, blood, peritoneal lavage, bone marrow, and spleen samples were collected for flow cytometry analyses of activated neutrophils (Ly6G+CD11b+), LDGs (CD15+CD14low), and NETs release (Sytox Green+).

Results: The pristane-induced mice group had a significantly increased number of blood activated neutrophils and LDGs as well as NETs released by these cells compared to the saline-injected control group and the basal values determined 12 days before the injection. The pristane group also had a significantly increased number of activated neutrophils, LDGs, and NETs released compared to the control group for the peritoneal lavage and bone marrow, except total cell count in spleen.

Conclusions: We demonstrated early changes in the innate immune response such as an increased number of activated neutrophils and LDGs and mainly increased NETosis in the pristane-induced mice model which may be considered as the primary event triggering lupus development.

MeSH terms

  • Animals
  • Disease Models, Animal*
  • Extracellular Traps* / immunology
  • Extracellular Traps* / metabolism
  • Female
  • Granulocytes / immunology
  • Granulocytes / metabolism
  • Lupus Erythematosus, Systemic* / chemically induced
  • Lupus Erythematosus, Systemic* / immunology
  • Lupus Erythematosus, Systemic* / pathology
  • Mice
  • Mice, Inbred BALB C*
  • Neutrophil Activation*
  • Neutrophils* / immunology
  • Spleen / immunology
  • Terpenes*

Substances

  • pristane
  • Terpenes

Grants and funding

This work was supported financial from the São Paulo Research Foundation – FAPESP [2017/02335-0]. The FAPESP is a public institution that promotes academic research of the government of the state of São Paulo, Brazil. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.