| Project/Area Number |
07457230
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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 |
Hematology
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
OHNO Ryuzo Hamamatsu University School of Medicine, Professor, 医学部, 教授 (70093002)
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| Co-Investigator(Kenkyū-buntansha) |
YANAGI Mitsuaki Hamamatsu University School of Medicine, Senior Resident, 医学部附属病院, 医員
TOBITA Tadasu Hamamatsu University School of Medicine, Senior Resident, 医学部附属病院, 医員
TAKESHITA Akihiro Hamamatsu University School of Medicine, Research Associate, 医学部附属病院, 助手 (00242769)
OHNISHI Kazunori Hamamatsu University School of Medicine, Assistant Professor, 医学部附属病院, 講師 (80252170)
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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 |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1995: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | MDS / erythropoietin receptor / thrombopoietin receptor / CD34 / CD38 / glycophorin A / glycoprotein IIb / IIIa / IIIa / エリスロポイエチンレセプター / Glycophorin-A |
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| Research Abstract |
In hematological diseases, abnormal expression of cytokine receptors and disorder of postreceptor signal transduction have been reported. However, little is known regarding the expression levels of cytokine receptors on cells in clinical samples because of the low level of expression and the difficulty in isolation of objective cells. In this study, we analyzed expression levels of erythropoietin (EPO) receptors and thrombopoietin (TPO) receptors on various phenotypic cells using a newly-devised non-isotopic ligand assay and evaluated its disorder in myelodysplastic syndromes (MDS). Some cell lines or bone marrow cells were incubated with biotinylated EPO or TPO followed by staining with streptavidine-RED670 conjugates and the fluorescence intensity was measured by a glow cytometer. The expression level of EPO receptors on cells was evaluated with the fluorescence intensity and the D balue calculated by the Kolmogorov-Smirnov statistics. By the simultaneous staining with anti-CD34, CD38, glycophorin A or glycoprotein IIb/IIIa, the expression levels of EPO receptors and TPO receptors on each phonotypic cells were analyzed. The specificity of this assay was confirmed by an experiment of cell lines EPO receptors were detected quantitatively in CD34 and/or glycophorin A positive fractions and TPO receptors were detected quantitatively in CD34 and/or glycoprotein IIb/IIIa positive fractions. The expression levels of both EPO receptors and TPO receptors on cells from MDS patients were decreased. In conclusion, the expression of EPO receptors on erythroid cells and TPO receptors on megakaryocytes were able to be detected quantitatively by non-isotopic ligand assay. This assay was specific and sensitive, and was performed easoly. The expression levels of both EPO receptors and TPO receptors on cells from MDS patients were decreased, and thus gene therapy introducting these receptor genes into MDS cells may be warranted.
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