中性粒细胞胞外陷阱蛋白质组学分析揭示了可能与DADA2患者树突状细胞介导的炎症相关的新途径。
NET Proteomic Profiling Reveals New Pathways Potentially Implicated in Dendritic Cell-Mediated Inflammation in DADA2 Patients.
作者信息
Signa Sara, Bartolucci Martina, Bonacini Martina, Bertoni Arinna, Del Zotto Genny, Corcione Anna, Petretto Andrea, Della Bella Silvia, Bertelli Roberta, Di Silvestre Dario, Lomagno Andrea, Mauri Pierluigi, Caorsi Roberta, Bruschi Maurizio, Balin Simone, Bocca Paola, Volpi Stefano, Catanoso Maria Grazia, Cafaro Alessia, Tripodi Gino, Pellottieri Lorenzo, Mavilio Domenico, Insalaco Antonella, Croci Stefania, Salvarani Carlo, Gattorno Marco, Schena Francesca
机构信息
UOC Reumatologia E Malattie Autoinfiammatorie, IRCCS Istituto Giannina Gaslini, Genoa, Italy.
Core Facilities-Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, Genoa, Italy.
出版信息
J Clin Immunol. 2025 Jun 16;45(1):106. doi: 10.1007/s10875-025-01888-w.
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.
目的
腺苷脱氨酶2缺乏症（DADA2）是一种自身炎症性疾病，其特征为系统性血管病变、中风和轻度免疫缺陷。最近，中性粒细胞胞外陷阱（NETosis）与腺苷脱氨酶2缺乏症的发病机制有关。为深入研究NETs对免疫系统的可能影响，我们对DADA2患者的NETs蛋白质组学特征进行了表征，并与健康对照（HD）和结节性多动脉炎（PAN）患者进行了比较。为确定NETs是否含有可能具有免疫原性的抗原，我们研究了用NETs体外刺激后树突状细胞的功能方面。
方法
招募了23名DADA2患者。我们通过成像流式细胞术分析NETosis。我们通过ELISA测定法评估血浆样本中的NETs残余物和脱氧核糖核酸酶（DNAse），而通过DNA消化评估DNAse活性。我们使用定量蛋白质组学方法和网络分析来鉴定6名DADA2患者、7名PAN患者和7名HD患者的NET蛋白和信号通路。我们通过流式细胞术分析循环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出自杀性NETosis显著增加。DNAse酶的水平或活性不正常。通过蛋白质组学分析，我们鉴定出1356种蛋白质，其中与正常和疾病对照的静息状态及用佛波酯（PMA）、腺苷和肿瘤坏死因子α（TNFα）刺激后的NETs相比，有100种蛋白质在DADA2 NETs中显著上调或下调。DADA2 NETs在刺激患者单核细胞来源的树突状细胞方面比正常NETs显著更有效。
结论
我们确定了DADA2 NETs与PAN/HD NETs相比有显著调节的不同信号通路。这种独特的蛋白质谱可能有助于激活DADA2中树突状细胞的炎症信号通路。
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