1998 Fiscal Year Final Research Report Summary
Expression and function of fusion regulatory protein-1/CD98 in monocytes
| Project/Area Number |
08671652
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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 |
Orthopaedic surgery
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| Research Institution | Mie University |
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Principal Investigator |
SUDO Akihiro Mie University, Hospital, Lecturer., 医学部附属病院, 講師 (60196904)
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| Project Period (FY) |
1996 – 1998
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| Keywords | Osteoclast / Monocyte / Fusion Regulatory Protein (FRP) -1 / CD98 / Monoclonal Antibody / c-src |
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| Research Abstract |
We have developed a new and simple system of human osteoclast formation by fusing peripheral blood monoocytes with anti-Fusion Regulatory Protein-1(FRP-1) mAb. When human blood monocytes were cultured in the presence of anti-FRP-1/CD98 mobs, po1ykaryocy5es began to appear at approximately 15 h and increased in size with time until l 3 to 4 days of incubation with anti-FRP-1 mAb. These fused cells showed positive staining in tartrate-resistant acid phosphatase (TRAP), possessed numerous calcitonin receptors(CTRs) and were capable of bone resorption. These results strongly suggests that anti-FRP-1antibody-induced multinucleated cells are osteoclasts. mRFJA expression of markers related to osteoclasts was analyzed during differentiation of osteoclasts from nonocytes. The mRNAs other than c-src nRNA were expressed in freshly isolated monocytes or monocytes incubated with control antibody or anti-FRP-1 mAbfor 14 days. Out of these mRNAs, cataepsin-K, CA II, nu -ATPase, N/NR, TRAP, OPN and c
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-fms mRNAs were expressed at higher levels in the osteoclost-like cells than in monocytes with control antibody. On the other hand, golectin-3 mRNA was expressed at lower levels in the osteoclast-like cells, and there was rosignificant difference in c-fos mRNA expression between the two.
c-src mRNA could not be detected in monocytes freshly isolated or incubated with control antibody. Intriguingly, expression of c-src mRNA was induced in monocytes by anti-FRP-1 nib and a detectable level of c-src mRNA expression was observed as early as 3 hours after anti-FRP-1 mAbs treotment, indicating that c-src is selectively induced by anti-FRP-1 nib treatment. The osteoclast-like cells and IleLa cellsas control cells were stained with anti-FRP-1 mobs and anti-c-src antibody followed by fluorescent-tagged secondary antibodies and subjected to a confocal laser microscopy. c-src and FRP-1were found to be mainly present around nucleus and were faintly stained in cytoplasm. In this study, FRP-1 and c-src were found to colocalize in osteoclasts, intimcrting that c-src is related toFRP-1-neediated signal trnasduction. Less
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