Biomedical Proteomics Research Group, Medical University Centre, Geneva, Switzerland.
Mol Cell Proteomics. 2010 Dec;9(12):2783-95. doi: 10.1074/mcp.M110.001008. Epub 2010 Aug 19.
Human African trypanosomiasis, or sleeping sickness, is a parasitic disease endemic in sub-Saharan Africa, transmitted to humans through the bite of a tsetse fly. The first or hemolymphatic stage of the disease is associated with presence of parasites in the bloodstream, lymphatic system, and body tissues. If patients are left untreated, parasites cross the blood-brain barrier and invade the cerebrospinal fluid and the brain parenchyma, giving rise to the second or meningoencephalitic stage. Stage determination is a crucial step in guiding the choice of treatment, as drugs used for S2 are potentially dangerous. Current staging methods, based on counting white blood cells and demonstrating trypanosomes in cerebrospinal fluid, lack specificity and/or sensitivity. In the present study, we used several proteomic strategies to discover new markers with potential for staging human African trypanosomiasis. Cerebrospinal fluid (CSF) samples were collected from patients infected with Trypanosoma brucei gambiense in the Democratic Republic of Congo. The stage was determined following the guidelines of the national control program. The proteome of the samples was analyzed by two-dimensional gel electrophoresis (n = 9), and by sixplex tandem mass tag (TMT) isobaric labeling (n = 6) quantitative mass spectrometry. Overall, 73 proteins were overexpressed in patients presenting the second stage of the disease. Two of these, osteopontin and β-2-microglobulin, were confirmed to be potential markers for staging human African trypanosomiasis (HAT) by Western blot and ELISA. The two proteins significantly discriminated between S1 and S2 patients with high sensitivity (68% and 78%, respectively) for 100% specificity, and a combination of both improved the sensitivity to 91%. The levels of osteopontin and β-2-microglobulin in CSF of S2 patients (μg/ml range), as well as the fold increased concentration in S2 compared with S1 (3.8 and 5.5 respectively) make the two markers good candidates for the development of a test for staging HAT patients.
非洲人类锥虫病，又称昏睡病，是一种寄生虫病，流行于撒哈拉以南非洲地区，通过采采蝇叮咬传播给人类。疾病的第一或血淋巴期与寄生虫在血液、淋巴系统和身体组织中的存在有关。如果患者未经治疗，寄生虫会穿过血脑屏障并侵入脑脊液和脑实质，从而引发第二或脑膜脑炎期。分期确定是指导治疗选择的关键步骤，因为用于 S2 的药物有潜在危险。目前的分期方法基于白细胞计数和脑脊液中锥虫的检测，缺乏特异性和/或敏感性。在本研究中，我们使用了几种蛋白质组学策略来发现具有分期非洲人类锥虫病潜力的新标志物。从刚果民主共和国感染布氏冈比亚锥虫的患者中采集脑脊液 (CSF) 样本。根据国家控制计划的指南确定分期。通过二维凝胶电泳 (n = 9) 和六重串联质量标签 (TMT) 等离 子标记定量质谱分析 (n = 6) 分析样品的蛋白质组。总体而言，在患有疾病第二阶段的患者中，有 73 种蛋白质表达上调。其中两种，骨桥蛋白和β-2-微球蛋白，通过 Western blot 和 ELISA 被确认为分期非洲人类锥虫病 (HAT) 的潜在标志物。这两种蛋白质对 S1 和 S2 患者的区分具有高灵敏度 (分别为 68%和 78%) 和 100%的特异性，并且两者的组合可将灵敏度提高到 91%。S2 患者脑脊液中骨桥蛋白和β-2-微球蛋白的水平 (μg/ml 范围) 以及与 S1 相比 S2 增加的倍数 (分别为 3.8 和 5.5) 使这两种标志物成为分期 HAT 患者检测方法的良好候选者。
