ELISA ASSAY OF CYTOKERATIN IN CEREBROSPINAL FLUID OF PATIENTS WITH INTRACRANIAL TUMORS

Research Project

Project/Area Number	63480327
Research Category	Grant-in-Aid for General Scientific Research (B)
Allocation Type	Single-year Grants
Research Field	Cerebral neurosurgery
Research Institution	NIIGATA UNIVERSITY
Principal Investigator	ONDA Kiyoshi NIIGATA UNIVERSITY MEDICAL HOSPITAL, ASSISTANT, 医学部附属病院, 助手 (90201409)
Co-Investigator(Kenkyū-buntansha)	TANAKA Ryuichi NIIGATA UNIVERSITY BRAIN RESEARCH INSTITUTE, PROFESSOR, 脳研究所, 教授 (30018816)
Project Period (FY)	1988 – 1989
Project Status	Completed (Fiscal Year 1989)
Budget Amount *help	¥2,100,000 (Direct Cost: ¥2,100,000) Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1988: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywords	Keratin / Monoclonal antibody / ELISA / Tumor marker / モノクロナル抗体 / サイトケラチン / モノクロナール抗体
Research Abstract	A monoclonal antibody (25G) against keratin intermediate filaments was prepared by fusing myelona cells with splenic lymphocytes from mice immunized with a cytoskeleton of established cell line from large cell lung carcinoma. The antibody was tested on ethanol and formalin fixed paraffin embedded human tissues respectively. This antibody showed wide spectrum of reactivity either normal or neoplastic epithelial tissues. No reaction was found in neuronal, glial and nesenchynal tissues and its tumors. Using this antibody, we developed avidin-biotin detection system for ELISA assay, by which, above 50ng/ml of keratin protein could be detected. ELISA assays of keratin protein in the cerebrospinal fluid (CSF) was carried out in patients with metastatic brain tumors, primary brain tumors and healthy controls. Elevated levels of CSF keratin protein were noted in 5 (29%) out of 17 metastatic brain tumors (2 squamous cell carcinoma, 2 adenocarcinona, 1 not verified). On the other hand, no elevated levels of CSF keratin protein were observed in all primary brain tumors (10 gliona, 5 malignant lymphoma, 2 nedduloblastona, 2 meningiona, 1 acoustic neurinona) and healthy controls. These findings indicated that determination of keratin proteins in CSF is valuable for differential diagnosis between primary and metastatic brain tumors.