NET Proteomic Profiling Reveals New Pathways Potentially Implicated in Dendritic Cell-Mediated Inflammation in DADA2 Patients.

IF 5.7 2区医学 Q1 IMMUNOLOGY

Journal of Clinical Immunology Pub Date : 2025-06-16 DOI:10.1007/s10875-025-01888-w

Sara Signa, Martina Bartolucci, Martina Bonacini, Arinna Bertoni, Genny Del Zotto, Anna Corcione, Andrea Petretto, Silvia Della Bella, Roberta Bertelli, Dario Di Silvestre, Andrea Lomagno, Pierluigi Mauri, Roberta Caorsi, Maurizio Bruschi, Simone Balin, Paola Bocca, Stefano Volpi, Maria Grazia Catanoso, Alessia Cafaro, Gino Tripodi, Lorenzo Pellottieri, Domenico Mavilio, Antonella Insalaco, Stefania Croci, Carlo Salvarani, Marco Gattorno, Francesca Schena

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引用次数: 0

Abstract

Purpose: Adenosine deaminase 2 Deficiency (DADA2) is an autoinflammatory disease characterized by systemic vasculopathy, strokes and mild immunodeficiency. Recently NETosis has been implicated in the pathogenesis of Deficiency of Adenosine Deaminase 2. To deep investigate the possible effects of NETs on the immune system we characterized proteomic profile of NETs from DADA2 as compared to HD and Polyarteritis Nodosa (PAN) patients. To determine if NETs contain possibly immunogenic antigens we study functional aspects on Dendritic Cells after in vitro stimulation with NETs.

Methods: Twenty-three DADA2 patients were enrolled. We analyzed NETosis by Imaging Flow Citometry. We evaluated NETs remnants and DNAse in the plasma samples by ELISA assay whereas DNAse activity by DNA digestion. We used quantitative proteomics approach and network analysis to identify NET proteins and pathways in 6 DADA2, 7 PAN and 7 HD. We analyzed circulating and monocyte-derived dendritic cells by flow cytometry.

Results: Neutrophils from DADA2 patients show a significant increased suicidal NETosis. DNAse enzymes were not normal in the level or activity. By proteomic analysis we identified 1356 proteins among which a hundred of proteins were significantly up or down-modulated in DADA2 NETs as compared to normal and disease control NETs in resting condition and after stimulation with PMA, Adenosine and TNFα. DADA2 NETs are significantly more efficient than normal NETs in stimulating patients' monocyte-derived dendritic cells.

Conclusion: We identified different pathways significantly modulated in DADA2 NETs versus PAN/HD NETs. This peculiar protein profile could contribute in activating inflammatory pathways in Dendritic cells in DADA2.

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NET蛋白质组学分析揭示了DADA2患者树突状细胞介导炎症的新途径。

目的：腺苷脱氨酶2缺乏症（DADA2）是一种以全身血管病变、中风和轻度免疫缺陷为特征的自身炎症性疾病。最近NETosis被认为与腺苷脱氨酶2缺乏的发病机制有关。为了深入研究NETs对免疫系统的可能影响，我们将DADA2患者的NETs与HD和结节性多动脉炎（PAN）患者的NETs进行了蛋白质组学分析。为了确定NETs是否含有可能的免疫原性抗原，我们研究了NETs体外刺激后树突状细胞的功能方面。方法：纳入23例DADA2患者。我们用血流成像法分析NETosis。我们用酶联免疫吸附法（ELISA）测定血浆样品中的NETs残体和DNA酶活性，用DNA酶切法测定DNA酶活性。我们使用定量蛋白质组学方法和网络分析鉴定了6个DADA2、7个PAN和7个HD的NET蛋白及其通路。我们用流式细胞术分析循环树突状细胞和单核细胞来源的树突状细胞。结果：DADA2患者的中性粒细胞显示自杀性NETosis显著增加。脱氧核糖核酸酶的水平和活性均不正常。通过蛋白质组学分析，我们鉴定出1356个蛋白，其中100个蛋白在静息状态和PMA、腺苷和TNFα刺激后，在DADA2 NETs中与正常和疾病对照的NETs相比显著上调或下调。DADA2 NETs在刺激患者单核细胞来源的树突状细胞方面明显比正常NETs更有效。结论：我们确定了DADA2 NETs与PAN/HD NETs中显著调节的不同通路。这种特殊的蛋白质谱可能有助于激活DADA2树突状细胞的炎症途径。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Clinical Immunology 医学-免疫学

CiteScore

12.20

自引率

9.90%

发文量

218

审稿时长

2 months

期刊介绍： The Journal of Clinical Immunology publishes impactful papers in the realm of human immunology, delving into the diagnosis, pathogenesis, prognosis, or treatment of human diseases. The journal places particular emphasis on primary immunodeficiencies and related diseases, encompassing inborn errors of immunity in a broad sense, their underlying genotypes, and diverse phenotypes. These phenotypes include infection, malignancy, allergy, auto-inflammation, and autoimmunity. We welcome a broad spectrum of studies in this domain, spanning genetic discovery, clinical description, immunologic assessment, diagnostic approaches, prognosis evaluation, and treatment interventions. Case reports are considered if they are genuinely original and accompanied by a concise review of the relevant medical literature, illustrating how the novel case study advances the field. The instructions to authors provide detailed guidance on the four categories of papers accepted by the journal.