| Project/Area Number |
07557154
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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 | 試験 |
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| Research Field |
Biological pharmacy
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| Research Institution | The 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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| Co-Investigator(Kenkyū-buntansha) |
YAMAOKA Takashi Sumitomo Pharmaceuticals, Research Center, Principal Investigator, 総合研究所, 主任研究員
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1996: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1995: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | Cell transplantation / Cellular trafficking / MMGL / Lectin / Experimental lung metastasis / Adoptive immunotherapy |
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| Research Abstract |
Cell transplantation is as important as organ transplantation as a subject of medical research. An important problem is how to target transplanted cells into appropriate organs or tissues. Recent progress revealed importance of specific recognition of carbohydrate ligands by endogenous lectins in physiological targeting of cells. In this study, we investigated an artificial targeting of cells into tumor sites using MMGL,which is a transmembrane calcium-type lectin found on tumoricidal macrophages, as a carbohydrate recognition device. We also tried to develop novel methods to analyze cellular targeting.
1. Interleukin 2-dependent mouse T cell line, CTLL-2 cells, were transfacted with an expression vector pCEP-4 containing full length cDNA insert encoding MMGL.A stable transfectant cell line, CTL-ML,was obtained. Cell surface expression of MMGL was demonstrated by mAb LOM-14 against MMGL that we developed.
2. The CTL-ML cells labeled with a fluorescent dye DiI were intravenously injected
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into mice with established lung metastases produced by mouse ovarian tumor OV2944-HM-1cells. Using sequential frozen sections of lungs, fluorescent cell numbers in the metastatic nodules were compared with those in the normal lung tissues under a fluorescence microscope. CTL-ML cells were found to selectively accumulate in the tumor tissue as compared with mock transfectant CTL-CEP cells.
3. For data analysis, image analyzer and confocal microscope were tested. Especially confocal microscope was useful for detection of labeled cells and measurement of sample area.
4. MMGL has an cytoplasmic motif which is involved in endocytic pathway. To display MMGL on cell surface more efficiently, we eliminated the motif to reduce down regulation by endocytosis. A stable transfectant CTL-YM,which has altered cytoplasmic motif AENL instead of YENL,was produced. Cell surface expression of MMGL was confirmed. We have not seen significant differences in the cellular accumulation into lung metastatic nodules between CTL-ML and CTL-YM at present. Less
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