| Project/Area Number |
17590489
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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 |
Laboratory medicine
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
TAKESHITA Kaori Hamamatsu University School of Medicine, University Hospital, Senior Resident (30397393)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHITA Akihiro Hamamatsu University School of Medicine, University Hospital, Associate Professor (00242769)
OHNISHI Kazunori Hamamatsu University School of Medicine, University Hospital, Professor (80252170)
MAEKAWA Masato Hamamatsu University School of Medicine, School of Medicine, Professor (20190291)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,710,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | cytokine / receptor / hormone / thyrotropin / flow cytometry / carbohydrate chain / mpl / 造血因子 / フローサイトメトリー |
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| Research Abstract |
Recently we have devised a non-isotopic ligand binding assay of G-CSF receptor, erythropoietin receptor and mpl with the flow cytometry using biotinylated cytokines and fluorescent dye labeled streptavidin. Also, we have enabled to analyze a very small amount of receptors quantitatively according to the cell surface antigen by multicolor flow cytometry even in the heterogeneous cell population. In this study, we examined the mpl expression level of the cell surface on acute leukemia cells, and also examined the possibility of applying the non-isotopic ligand binding assay with the flow cytometry to the hormone receptor assay.
We analyzed the number of surface mpl on leukemic cells obtained from patients of acute leukemia. Although mpl was expected to be expressed in the cases of FAB classification M7 from its characteristics, it was expressed on the leukemic cells broadly from MO to M7. It was also expressed on ALL cells. When _3H-tymidine incorporation were performed, those leukemic ce
… More
lls had proliferation potency in some cases. However, there was considerably variation of each specimen and was not able to identify a cause of the variation. The correlation with the surface antigen expression was not found, too. For development application to hormone receptor analysis, we examined the possibility of analysis of the thyrotropin (TSH) receptor expression by using recombinant human TSH (rhTSH) and cell lines which were expressing TSH receptors. We tried biotinylation of the rhTSH like the above-mentioned cytokines, but enough biotinylation was not attained. So improvement of biotinylated efficiency was thought to be.necessary. Also, we tried to label the fluorescent dye to the rhTSH directly, but the amplification of the fluorescence intensity by the specific binding to a TSH receptor of the fluorescent labeling TSH was insufficient. Therefore, in regard to a quantitative analysis of TSH receptor by the flow cytometer, it should be considered about the more effective intensification of biotinylation or fluorescent labeling of the rhTSH, and it was thought that the improvement of the detectability was indispensable. Less
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