Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration
Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration
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DOI：
10.1016/j.expneurol.2008.06.004
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发表时间：
2008-10-01
影响因子：
5.3
通讯作者：
Schlossmacher, Michael G.
中科院分区：
文献类型：
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作者：
Mollenhauer, Brit;Cullen, Valerie;Schlossmacher, Michael G.
由于α-突触核蛋白（α S）在大脑中的积累是帕金森病（PD）和相关疾病的标志，我们研究了其在人类脑脊液（CSF）中的发生。在从神经健康供体收集的CSF的亲和富集和胰蛋白酶消化之后，我们通过质谱法鉴定了几种α S衍生肽。我们的浓度相当于CSF蛋白质组的< 0.001%。然后，我们建立，验证和优化了一种酶联免疫吸附试验（ELISA），以测量未浓缩CSF中的总α S水平。在一项100名活体供者的横断面研究中，我们检查了临床诊断为晚期PD、路易体痴呆（DLB）、阿尔茨海默病（AD）和一组非神经退行性疾病对照（NCO）的受试者的无细胞CSF样本。在这四组中，CSF α S浓度范围为0.8 - 16.2 pg/μ l。患有原发性突触核蛋白病的供体（PD，DLB：n=57）的平均CSF aS值低于其他两组（AD，NCO：n=35; p=0.025）。相比之下，存活的克雅氏病患者显示CSF α S水平显著升高（n=8;平均值为300 pg/mu l; p < 0.001）。我们的结果明确证实了成人CSF中存在aS。在采用新型ELISA的第一项可行性研究中，我们发现与突触核蛋白病、PD和DLB相关的帕金森综合征受试者的CSF α S浓度相对较低。在明确的朊病毒病病例中，我们记录到由于细胞快速死亡导致的总CSF α S显著升高。我们的研究结果可能有助于未来在神经退行性疾病中的生物标志物探索，并促进靶点验证研究。(c)2008年爱思唯尔公司All rights reserved.
Because accumulation of alpha-synuclein (alpha S) in the brain is a hallmark of Parkinson disease (PD) and related disorders, we examined its Occurrence in human cerebrospinal fluid (CSF). Following affinity enrichment and trypsin digestion of CSF collected from a neurologically healthy donor, we identified several alpha S-derived peptides by mass spectrometry. The concentration of us amounted to < 0.001% of the CSF proteome. We then built, validated and optimized a sandwich-type, enzyme-linked immunoadsorbent assay (ELISA) to measure total alpha S levels in unconcentrated CSF. In a cross-sectional study of 100 living donors, we examined cell-free CSF samples from subjects clinically diagnosed with advanced PD, dementia with Lewy bodies (DLB), Alzheimer disease (AD), and a group of non-neurodegenerative disease controls (NCO). In these four groups the CSF alpha S concentrations ranged from 0.8 to 16.2 pg/mu l. Mean CSF aS values were lower in donors with a primary synucleinopathy (PD, DLB: n=57) than in the other two groups (AD, NCO: n=35; p=0.025). By contrast, living Creutzfeldt-Jakob disease patients showed markedly elevated CSF alpha S levels (n=8; mean, 300 pg/mu l; p < 0.001). Our results unequivocally confirm the presence of aS in adult human CSF. In a first feasibility study employing a novel ELISA, we found relatively low CSF alpha S concentrations in subjects with parkinsonism linked to synucleinopathy, PD and DLB. In definite prion disease cases, we recorded a marked rise in total CSF alpha S resulting from rapid cell death. Our results will likely aid future biomarker explorations in neurodegenerative conditions and facilitate target validation studies. (c) 2008 Elsevier Inc. All rights reserved.