| Project/Area Number |
12670500
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Nagasaki University Hospital |
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Principal Investigator |
MAKIYAMA Kazuya Nagasaki University Hospital Associate Professor, 医学部・附属病院, 助教授 (70112381)
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| Co-Investigator(Kenkyū-buntansha) |
TSUZIMURA Noriyuki Sakamoto Bio Co., Ltd. Chief Researcher, 主任研究員
ISOMOTO Hazime Nagasaki University Hospital Lecturer, 医学部・附属病院, 助手 (90322304)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | ulcerative colitis / eosinophil cationic protein: ECP / polypeptides / polyclonal antibody / eosinophil cationic protein |
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| Research Abstract |
To investigate the role of activated eosinophils in the inflammatory process of ulcerative colitis, we prepared antibodies against polypeptides of a human eosinophil granule protein, eosinophil cationic protein (ECP), in attempt to purify ECP.
1. The following four ECP polypeptides were selected based on the human ECP amino acid sequence reported by Barker et al.: ECP1, CRYADRPGRRFVVA; ECP2, CDNRDPRDSPRYPVV; ECP3, CTIAMRAINNYRWRC; and ECP4, TRAQWFAIQHISLNPPR.
2. Rabbits were immunized with ECP polypeptide antigens, and anti-human ECP antibodies that reacted with synthetic peptides even the rabbit antisera were diluted to 1/10,000 were obtained. 3. None of the acquired anti-human ECP antibodies reacted as anti-ECP antiserum in an ECP assay system using Pharmacia ECP RIA kit. 4. Anti-human ECP2 antibody and anti-human ECP3 antibody reacted with eosinophils and monocytes in immunohistochemical staining.
The above findings suggested that although it is necessary to confirm whether the acquired anti-ECP antibodies react with human ECP, at least the ECP2 and ECP3 amino acid sequence regions have immunological reactivity. The anti-inflammatory effect of the anti-human ECP antibodies should be investigated using an experimental animal model.
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