A proteomic repertoire of autoantigens identified from the classic autoantibody clinical test substrate HEp-2 cells.
A proteomic repertoire of autoantigens identified from the classic autoantibody clinical test substrate HEp-2 cells.
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DOI：
10.1186/s12014-020-09298-3
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发表时间：
2020
影响因子：
3.8
通讯作者：
Roehrl MH
中科院分区：
文献类型：
--
作者：
Wang JY;Zhang W;Rho JH;Roehrl MW;Roehrl MH
自身抗体是自身免疫性疾病的标志。用患者血清间接免疫荧光染色HEp-2细胞进行自身抗体筛选是目前临床实践中的标准方法。自身免疫性疾病的鉴别诊断是基于常见的核和细胞质染色模式。在这项研究中，我们尝试使用一种独特的蛋白质组学ds亲和富集策略从HEp-2细胞中鉴定尽可能多的自身抗原。培养并裂解HEp-2细胞。从细胞裂解液中提取总蛋白并用DS-Sepharose树脂进行分离。用盐梯度洗脱蛋白质，收集低至高亲和力的部分，并用质谱法进行测序。通过文献文本挖掘来验证每个蛋白的自身抗原性。对所有鉴定的蛋白进行蛋白相互作用网络和通路分析。本研究鉴定了107种具有低至高ds亲和力的蛋白。其中，78个已被证实为自身抗原，并有先前的报道作为自身抗体的靶点，而29个可能是潜在的自身抗原，但尚未得到证实。107个蛋白中82个位于细胞核，15个位于有丝分裂细胞周期，这可能与HEp-2试验中细胞核和有丝分裂染色模式的优势相对应。HEp-2染色中存在55种囊泡相关蛋白和12种核糖核蛋白颗粒蛋白，这可能是HEp-2染色中出现各种斑点的原因。还有32种蛋白质与细胞骨架相关。蛋白质网络分析表明，这些蛋白质之间的相互作用明显多于随机组合，其中排名前3位的网络分别是mRNA代谢过程调节、细胞凋亡和DNA构象改变。这项研究为未来的研究提供了已证实的和潜在的自身抗原的蛋白质组学库，并且这些发现与自身抗原的机制一致：自身分子如何与DS形成分子复合物以引发自身免疫。我们的数据有助于自身免疫的分子病因学，并可能加深我们对自身免疫性疾病的理解。
Autoantibodies are a hallmark of autoimmune diseases. Autoantibody screening by indirect immunofluorescence staining of HEp-2 cells with patient sera is a current standard in clinical practice. Differential diagnosis of autoimmune disorders is based on commonly recognizable nuclear and cytoplasmic staining patterns. In this study, we attempted to identify as many autoantigens as possible from HEp-2 cells using a unique proteomic DS-affinity enrichment strategy. HEp-2 cells were cultured and lysed. Total proteins were extracted from cell lysate and fractionated with DS-Sepharose resins. Proteins were eluted with salt gradients, and fractions with low to high affinity were collected and sequenced by mass spectrometry. Literature text mining was conducted to verify the autoantigenicity of each protein. Protein interaction network and pathway analyses were performed on all identified proteins. This study identified 107 proteins from fractions with low to high DS-affinity. Of these, 78 are verified autoantigens with previous reports as targets of autoantibodies, whereas 29 might be potential autoantigens yet to be verified. Among the 107 proteins, 82 can be located to nucleus and 15 to the mitotic cell cycle, which may correspond to the dominance of nuclear and mitotic staining patterns in HEp-2 test. There are 55 vesicle-associated proteins and 12 ribonucleoprotein granule proteins, which may contribute to the diverse speckled patterns in HEp-2 stains. There are also 32 proteins related to the cytoskeleton. Protein network analysis indicates that these proteins have significantly more interactions among themselves than would be expected of a random set, with the top 3 networks being mRNA metabolic process regulation, apoptosis, and DNA conformation change. This study provides a proteomic repertoire of confirmed and potential autoantigens for future studies, and the findings are consistent with a mechanism for autoantigenicity: how self-molecules may form molecular complexes with DS to elicit autoimmunity. Our data contribute to the molecular etiology of autoimmunity and may deepen our understanding of autoimmune diseases.
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影响因子：
6.8
作者：
Braunschweig D;Krakowiak P;Duncanson P;Boyce R;Hansen RL;Ashwood P;Hertz-Picciotto I;Pessah IN;Van de Water J
通讯作者：
Van de Water J
影响因子：
5.5
作者：
Frampton, G;Moriya, S;Murphy, JJ
通讯作者：
Murphy, JJ
影响因子：
6.1
作者：
Gajbhiye, R.;Sonawani, A.;Khole, V.
通讯作者：
Khole, V.
影响因子：
158.5
作者：
Arbuckle, MR;McClain, MT;Harley, JB
通讯作者：
Harley, JB
影响因子：
3.3
作者：
Cid, C.;Regidor, I.;Alcazar, A.
通讯作者：
Alcazar, A.