Carrasco Solange, Liphaus Bernadete L, Peixoto Tatiana Vasconcelos, Lima Thais Martins, Ariga Sueli Kunimi Kubo, Jesus Queiroz Zelita Aparecida, de Matos Lobo Thays, Catanozi Sergio, Rodrigues Letícia Gomes, Filho Antônio Santos, Teodoro Walcy Rosolia, Velosa Ana Paula Pereira, Levy Débora, Soriano Francisco Garcia, Goldenstein-Schainberg Cláudia
Laboratório de Imunologia Celular (LIM-17), Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil.
Laboratório de Pediatria Clínica (LIM-36), Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil.
PLoS One. 2025 Jan 3;20(1):e0306943. doi: 10.1371/journal.pone.0306943. eCollection 2025.
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.
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+).
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.
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.
中性粒细胞胞外诱捕网形成被认为是自身抗原的一个重要来源。因此，我们推测， pristane诱导的狼疮小鼠模型是否会出现中性粒细胞的早期激活、低密度粒细胞（LDG）的存在以及中性粒细胞胞外诱捕网（NET）的释放，而这些可能导致狼疮表型的发展。
给12只雌性野生型Balb/c小鼠腹腔注射pristane（n = 6；pristane组）或生理盐水（n = 6；对照组）。注射后5天，收集血液、腹腔灌洗液、骨髓和脾脏样本，用于对活化中性粒细胞（Ly6G+CD11b+）、LDG（CD15+CD14low）和NET释放（Sytox Green+）进行流式细胞术分析。
与注射生理盐水的对照组以及注射前12天测定的基础值相比，pristane诱导的小鼠组血液中活化中性粒细胞和LDG的数量以及这些细胞释放的NET显著增加。除脾脏中的总细胞数外，pristane组腹腔灌洗液和骨髓中活化中性粒细胞、LDG的数量以及释放的NET也比对照组显著增加。
我们证明了在pristane诱导的小鼠模型中先天性免疫反应的早期变化，如活化中性粒细胞和LDG数量增加，主要是NETosis增加，这可能被视为引发狼疮发展的主要事件。
