Cappelletti Chiara, Eriksson Anna, Brorson Ina Skaara, Leikfoss Ingvild S, Kråbøl Oda, Høgestøl Einar August, Vitelli Valeria, Mjaavatten Olav, Harbo Hanne F, Berven Frode, Bos Steffan D, Berge Tone
Department of Mechanical, Electronics and Chemical Engineering, Faculty of Technology, Art and Design, OsloMet-Oslo Metropolitan University, Postboks 4, St. Olavs Plass, 0130, Oslo, Norway.
Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway.
Clin Proteomics. 2022 Jul 5;19(1):23. doi: 10.1186/s12014-022-09361-1.
Multiple sclerosis (MS) is an autoimmune, neurodegenerative disorder with a strong genetic component that acts in a complex interaction with environmental factors for disease development. CD4 T cells are pivotal players in MS pathogenesis, where peripherally activated T cells migrate to the central nervous system leading to demyelination and axonal degeneration. Through a proteomic approach, we aim at identifying dysregulated pathways in activated T cells from MS patients as compared to healthy controls.
CD4 T cells were purified from peripheral blood from MS patients and healthy controls by magnetic separation. Cells were left unstimulated or stimulated in vitro through the TCR and costimulatory CD28 receptor for 24 h prior to sampling. Electrospray liquid chromatography-tandem mass spectrometry was used to measure protein abundances.
Upon T cell activation the abundance of 1801 proteins was changed. Among these proteins, we observed an enrichment of proteins expressed by MS-susceptibility genes. When comparing protein abundances in T cell samples from healthy controls and MS patients, 18 and 33 proteins were differentially expressed in unstimulated and stimulated CD4 T cells, respectively. Moreover, 353 and 304 proteins were identified as proteins exclusively induced upon T cell activation in healthy controls and MS patients, respectively and dysregulation of the Nur77 pathway was observed only in samples from MS patients.
Our study highlights the importance of CD4 T cell activation for MS, as proteins that change in abundance upon T cell activation are enriched for proteins encoded by MS susceptibility genes. The results provide evidence for proteomic disturbances in T cell activation in MS, and pinpoint to dysregulation of the Nur77 pathway, a biological pathway known to limit aberrant effector T cell responses.
多发性硬化症（MS）是一种自身免疫性神经退行性疾病，具有很强的遗传成分，在疾病发展过程中与环境因素发生复杂的相互作用。CD4 T细胞是MS发病机制中的关键参与者，外周激活的T细胞迁移到中枢神经系统，导致脱髓鞘和轴突退化。通过蛋白质组学方法，我们旨在确定与健康对照相比，MS患者活化T细胞中失调的信号通路。
通过磁珠分选从MS患者和健康对照的外周血中纯化CD4 T细胞。在取样前，细胞不进行刺激或通过TCR和共刺激CD28受体在体外刺激24小时。采用电喷雾液相色谱-串联质谱法测量蛋白质丰度。
T细胞活化后，1801种蛋白质的丰度发生了变化。在这些蛋白质中，我们观察到MS易感基因表达的蛋白质富集。比较健康对照和MS患者T细胞样本中的蛋白质丰度，分别有18种和33种蛋白质在未刺激和刺激的CD4 T细胞中差异表达。此外，分别在健康对照和MS患者中鉴定出353种和304种仅在T细胞活化后诱导的蛋白质，并且仅在MS患者的样本中观察到Nur77信号通路的失调。
我们的研究强调了CD4 T细胞活化对MS的重要性，因为T细胞活化后丰度发生变化的蛋白质富含MS易感基因编码的蛋白质。结果为MS中T细胞活化的蛋白质组学紊乱提供了证据，并指出Nur77信号通路失调，这是一种已知可限制异常效应T细胞反应的生物学途径。
