Department of Immunogenetics, Medical University of Lodz, Lodz, Poland.
Department of Periodontology and Oral Mucosal Diseases, Medical University of Lodz, Lodz, Poland.
Front Immunol. 2022 Jun 10;13:906311. doi: 10.3389/fimmu.2022.906311. eCollection 2022.
Neutrophils are a heterogenous population capable of both antimicrobial functions and suppressor ones, however, no specific pattern of transcription factors controlling this plasticity has been identified. We observed rapid changes in the neutrophil status after stimulation with LPS, pre-activating concentration of TNF-α, or IL-10. Chromatin immunoprecipitation sequencing (ChIP-Seq) analysis of histone H3K4me3 allowed us to identify various transcriptional start sites (TSSs) associated with plasticity and heterogeneity of human neutrophils. Gene Ontology analysis demonstrated great variation within target genes responsible for neutrophil activation, cytokine production, apoptosis, histone remodelling as well as NF-κB transcription factor pathways. These data allowed us to assign specific target genes positioned by H3K4me3-marked histone with a different pattern of gene expression related to NF-κB pathways, apoptosis, and a specific profile of cytokines/chemokines/growth factors realised by neutrophils stimulated by LPS, IL-10, or TNF-α. We discovered IL-10-induced apoptotic neutrophils being transcriptionally active cells capable of switching the profile of cytokines/chemokines/growth factors desired in resolving inflammation non-canonical NF-κB pathway with simultaneous inhibition of canonical NF-κB pathway. As apoptotic/suppressive neutrophils induced by IL-10 positioning genes within H3K4me3-marked histone were transcriptionally active, newly described DNA binding sites can be considered as potential targets for immunotherapy. H3K4me3 histone ChIP-Seq analysis reveals molecular drivers critical for switching neutrophils from their pro- to anti-inflammatory properties.
中性粒细胞是一个具有异质性的群体，既能发挥抗菌功能，也能发挥抑制功能，但目前尚未确定控制这种可塑性的特定转录因子模式。我们观察到，在受到 LPS、TNF-α 或 IL-10 预激活浓度刺激后，中性粒细胞状态迅速发生变化。组蛋白 H3K4me3 的染色质免疫沉淀测序（ChIP-Seq）分析使我们能够识别与人类中性粒细胞的可塑性和异质性相关的各种转录起始位点（TSS）。基因本体论分析表明，负责中性粒细胞激活、细胞因子产生、凋亡、组蛋白重塑以及 NF-κB 转录因子途径的靶基因存在很大差异。这些数据使我们能够为特定的靶基因分配位置，这些基因由 H3K4me3 标记的组蛋白所标记，与 NF-κB 途径、凋亡以及由 LPS、IL-10 或 TNF-α 刺激的中性粒细胞实现的特定细胞因子/趋化因子/生长因子特征相关的表达模式不同。我们发现，IL-10 诱导的凋亡中性粒细胞是具有转录活性的细胞，能够在炎症消退过程中切换所需的细胞因子/趋化因子/生长因子特征，而非经典 NF-κB 途径同时抑制经典 NF-κB 途径。由于 IL-10 诱导的凋亡/抑制性中性粒细胞在 H3K4me3 标记的组蛋白内定位基因具有转录活性，新描述的 DNA 结合位点可被视为免疫治疗的潜在靶点。H3K4me3 组蛋白 ChIP-Seq 分析揭示了将中性粒细胞从促炎状态切换为抗炎状态的关键分子驱动因素。
