| Project/Area Number |
05671813
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | UNIVERSITY OF TOKYO |
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Principal Investigator |
IMAI Yasuyuki Univ.of Tokyo, Fac.Pharmaceu.Sci., Research Associate, 薬学部, 助手 (80160034)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Macrophages / MMGL / Lectin / Monoclonal antibody / Cancer metastasis / Trafficking / モノクロナル抗体 / リンパ節 |
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| Research Abstract |
Aim of this project is to elucidate biological roles of mouse macrophage C-type lectin specific for galactose and N-acetylgalactosamine (MMGL) and to study a possible contribution of this molecule on cellular trafficking of normal and malignant cells. In order to establish topographical information on localization of MMGL-positive macrophages, we produced rat monoclonal antibodies (mAbs) specific for MMGL.We developed a novel carbohydrate binding assay based on an ELISA format to facilitate mAb screening. We systematically screened for sites of expression of MMGL using these mAbs by means of immunoblot and immunohistological analyzes, and found selective localization of MMGL-positive macrophages in connective tissue. We also detected MMGL-positive macrophages infiltrating in the metastatic nodules produced by metastatic mouse ovarian tumor cells in lung. Furthermore, we also demonstrated that MMGL is responsible for the recognition of truncated O-linked sugar chains displayd on tumor cells. In order to study possible contribution of MMGL to cellular trafficking as a cellular receptor, we transfected cDNA encoding MMGL to CTLL-2 mouse T cell line and asked whether properties of cellular trafficking could be changed. The expression of MMGL molecules was confirmed by immunoblot and flow-cytometry using mAbs. We found preferential localization of the MMGL-transfected cells within the metastatic noduled of mouse ovarian tumor cells upon injection of these cells labeled with fluorescent probes.
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