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.
The purpose of this Phase I application is to develop an in vitro test for identifying neuronal injury associated with traumatic brain damage.
