CSF MAP-2 IN TRAUMATIC BRAIN INJURY

描述：（申请人摘要）本提案的目的是开发一种用于定量急性神经元变性的实验室测试。这种测试是需要几个临床适应症，包括创伤性脑损伤（TBI）以及分期患者的神经保护剂的药物试验。拟议的实验室测试将量化微管相关蛋白2（MAP-2）的水平。MAP-2是主要定位于神经元细胞体和树突中的神经元特异性蛋白。在正常情况下，MAP-2不从神经元释放。然而，我们的初步蛋白质印迹研究表明，神经元损伤后（TBI患者）脑脊液（CSF）MAP-2水平升高，而对照CSF中的MAP-2水平检测不到。拟议的研究将提高单克隆抗体（单克隆抗体），识别在人类CSF中发现的MAP-2的形式。将产生抗从CSN创伤脑纯化的MAP-2的单克隆抗体，并针对来自TBI患者的CSF和对照CSF筛选杂交瘤，以确保单克隆抗体识别CSF MAP-2。这些MAP-2单克隆抗体将用于开发CSF MAP-2 ELISA。开发的MAP-2 ELISA将用于筛选来自TBI患者（N=30）和对照（N=61）的CSF，所述对照包括患有脱髓鞘疾病、脑积水、背痛和偏头痛的患者。通过比较TBI患者住院期间的CSF MAP-2水平与格拉斯哥昏迷量表评分，确定患者临床改善与CSF MAP-2水平之间的关系。将确定初始CSF MAP-2水平预测出院时TBI患者的临床结果（格拉斯哥结果评分）的能力。拟定商业应用：该I期申请的目的是开发用于鉴定与创伤性脑损伤相关的神经元损伤的体外测试。

DESCRIPTION: (Applicant's abstract) The objective of this proposal is to develop a laboratory test for quantifying acute neuronal degeneration. Such a test is needed for several clinical indications including traumatic brain injury (TBI) as well as staging patients for drug trials of neuroprotective agents. The proposed laboratory test will quantify levels of microtubule-associated protein 2 (MAP-2). MAP-2 is a neuron-specific protein localized primarily in neuronal cell bodies and dendrites. Under normal conditions MAP-2 is not released from neurons. However, our preliminary Western blot studies indicate that after neuronal injury (TBI patients) cerebrospinal fluid (CSF) MAP-2 levels are elevated while MAP-2 levels in control CSF were not detectable. The proposed studies will raise monoclonal antibodies (Mabs) that recognize the form of MAP-2 found in human CSF. Mabs will be raised against MAP-2 purified form CSN trauma brain and hybridomas screened against CSF from TBI patients and control CSF to insure that Mabs recognize CSF MAP-2. These MAP-2 Mabs will be employed to develop a CSF MAP-2 ELISA. The developed MAP-2 ELISA will be used to screen CSF from TBI patients (N=30) and controls (N=61) including patients with a demyelinating disease, hydrocephalus, back pain and migraine. The relationship between patient clinical improvement and CSF MAP-2 levels will be determined by comparing CSF MAP-2 levels with the Glasgow Coma Scale scores of TBI patients during hospitalization. The ability of initial CSF MAP-2 levels to predict the clinical outcome (Glasgow Outcome Score) of TBI patients at discharge will be determined. PROPOSED COMMERCIAL APPLICATION: The purpose of this Phase I application is to develop an in vitro test for identifying neuronal injury associated with traumatic brain damage.