Lind Anne-Li, Wu Di, Freyhult Eva, Bodolea Constantin, Ekegren Titti, Larsson Anders, Gustafsson Mats G, Katila Lenka, Bergquist Jonas, Gordh Torsten, Landegren Ulf, Kamali-Moghaddam Masood
Department of Surgical Sciences, Anaesthesiology and Intensive Care and Uppsala Berzelii Technology Center for Neurodiagnostics, Uppsala University, Uppsala, Sweden.
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
PLoS One. 2016 Feb 25;11(2):e0149821. doi: 10.1371/journal.pone.0149821. eCollection 2016.
The objective of this study was to develop and apply a novel multiplex panel of solid-phase proximity ligation assays (SP-PLA) requiring only 20 μL of samples, as a tool for discovering protein biomarkers for neurological disease and treatment thereof in cerebrospinal fluid (CSF). We applied the SP-PLA to samples from two sets of patients with poorly understood nervous system pathologies amyotrophic lateral sclerosis (ALS) and neuropathic pain, where patients were treated with spinal cord stimulation (SCS). Forty-seven inflammatory and neurotrophic proteins were measured in samples from 20 ALS patients and 15 neuropathic pain patients, and compared to normal concentrations in CSF from control individuals. Nineteen of the 47 proteins were detectable in more than 95% of the 72 controls. None of the 21 proteins detectable in CSF from neuropathic pain patients were significantly altered by SCS. The levels of the three proteins, follistatin, interleukin-1 alpha, and kallikrein-5 were all significantly reduced in the ALS group compared to age-matched controls. These results demonstrate the utility of purpose designed multiplex SP-PLA panels in CSF biomarker research for understanding neuropathological and neurotherapeutic mechanisms. The protein changes found in the CSF of ALS patients may be of diagnostic interest.
本研究的目的是开发并应用一种新型的多重固相邻近连接分析法（SP-PLA），该方法仅需20微升样本，作为发现脑脊液（CSF）中神经疾病蛋白质生物标志物及其治疗方法的工具。我们将SP-PLA应用于两组患有神经系统疾病（肌萎缩侧索硬化症（ALS）和神经性疼痛）且接受脊髓刺激（SCS）治疗的患者样本。对20例ALS患者和15例神经性疼痛患者的样本进行了47种炎症和神经营养蛋白的检测，并与对照个体脑脊液中的正常浓度进行比较。47种蛋白中的19种在72名对照者的95%以上可检测到。在神经性疼痛患者脑脊液中可检测到的21种蛋白中，没有一种因SCS而发生显著变化。与年龄匹配的对照组相比，ALS组中卵泡抑素、白细胞介素-1α和激肽释放酶-5这三种蛋白的水平均显著降低。这些结果证明了专门设计的多重SP-PLA分析板在脑脊液生物标志物研究中用于理解神经病理和神经治疗机制的实用性。在ALS患者脑脊液中发现的蛋白质变化可能具有诊断意义。
