2003 Fiscal Year Final Research Report Summary
PRODUCTION OF SPECIFIC MONOCLONAL ANTIBODY FOR CALPAIN CLEAVAGE SITES AND CLINICAL APPLICATION
| Project/Area Number |
14370460
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | GIFU UNIVERSITY |
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Principal Investigator |
SHIMIZU Katsuji GIFU UNIVERSITY, FACULTY OF MEDICINE, PROFESSOR, 医学部, 教授 (90170969)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Yoshiki GIFU UNIVERSITY, FACULTY OF MEDICINE, ASSISTANT, 医学部, 助手 (10313884)
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| Project Period (FY) |
2002 – 2003
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| Keywords | CALPAIN / CARTILAGE / PROTEOGLYCAN / ARTHRITIS |
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| Research Abstract |
Our previous reports revealed that calcium-dependent neutral cystein proteinase (calpain) is released from synovial and cartilage tissue to synovial fluid in osteoarthritis (OA) and rheumatoid arthritis (RA), and has strong proteoglycanase activity in vitro. These findings suggest that calpain plays an important role in destruction of the extra-cellular matrix (ECM) of cartilage in OA and RA suffers. Result of analyzing calpain cleavage sites on proteoglycan, we detected that calpin has many specific cleavage sites on proteoglycan core protein within articular cartilage ECM and in order of the cleavage sites sequences are different from another kinds of proteoglycanases sites.
Now we succeeded in production of monoclonal antibody which recognized only specific cleavage site on proteoglycan core protein by calpain, and demonstrated directly that the calain plays a role in destruction of the extra-cellular matrix (ECM) of cartilage.
Furthermore, I used the monoclonal antibody which specifically recognized degradation product by calpain to examine influence of inflammatory cytokine in bovine cartilaginous tissue culture system.
As a result of the pilot study, the quantity of calpain and the quantity of fragment by calpain cleavage were released from intracellular to the extracellular by adding inflammatory cytokine increased.
We developed one of the monoclonal antibody which specifically recognized degradation product of calpain and proved that calpain participated in articular cartilage destruction in arthritis.
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