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Neutrophil Extracellular Traps protein composition is specific for patients with Lupus nephritis and includes methyl-oxidized αenolase (methionine sulfoxide 93)

机译:中性粒细胞胞外陷阱蛋白的成分特定于狼疮肾炎患者包括甲基氧化的α-烯醇化酶(甲硫氨酸亚砜93)

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摘要

NETs constitute a network of DNA and proteins released by neutrophils in response to infectious and immunologic triggers. NET proteins are recognized as autoantigens in ANCA vasculitis; limited knowledge is available in other autoimmune pathologies. The composition of NETs produced ex vivo by resting and Phorbol-myristate acetate (PMA) stimulated neutrophils was analyzed by high-throughput Fusion Orbitrap technology in 16 patients with Systemic Lupus Erythematosus/Lupus nephritis (9 SLE/7 LN) and in 11 controls. Seven-hundred proteins were characterized and specific fingerprints discriminated LN from SLE. We focused on methyl-oxidized αenolase (methionine sulfoxide 93) that was markedly increased in NETs from LN and was localized in NET filaments in tight connection and outlying DNA. The isotype of anti-αenolase antibodies was IgG2 in LN and IgG4 in other autoimmune glomerulonephritis (Membranous Nephropathy, MN); serum anti-αenolase IgG2 were higher in LN than in SLE and absent in MN. The same IgG2 antibodies recognized 5 epitopes of the protein one containing methionine sulphoxide 93. In conclusion, specific NET protein fingerprints characterize different subsets of SLE; methyl-oxidized αenolase is over-expressed in LN. Circulating anti-αenolase IgG2 recognize the oxidized epitope and are high in serum of LN patients. Post-translational modified NET proteins contribute to autoimmunity in patients with LN.
机译:NET构成了由中性粒细胞释放的DNA和蛋白质网络,以响应感染和免疫触发。 NET蛋白被认为是ANCA血管炎的自身抗原。其他自身免疫性病理学知识有限。通过高通量融合Orbitrap技术分析了16例系统性红斑狼疮/狼疮性肾炎(9 SLE / 7 LN)患者和11例对照中静息和醋酸肉豆蔻酸乙酸酯(PMA)刺激的中性粒细胞离体产生的NET的组成。对700种蛋白质进行了表征,并用特异性指纹图谱从SLE中区分了LN。我们集中研究了甲基氧化的α-烯醇酶(甲硫氨酸亚砜93),该酶在LN的NETs中显着增加,并定位在NET细丝中紧密连接且位于外围的DNA中。抗α-烯醇化酶抗体的同种型是LN中的IgG2和其他自身免疫性肾小球肾炎(膜性肾病,MN)中的IgG4。 LN患者的血清抗α-烯醇酶IgG2高于SLE患者,而MN患者则不存在。相同的IgG2抗体可识别含有甲硫氨酸亚砜93的蛋白质的5个表位。总之,特定的NET蛋白指纹图谱可表征SLE的不同亚群。甲基氧化的α-烯醇化酶在LN中过表达。循环的抗α-烯醇化酶IgG2识别氧化表位,并且在LN患者的血清中含量很高。翻译后修饰的NET蛋白有助于LN患者的自身免疫。

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