| Project/Area Number |
10672170
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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 | The University of Tokyo |
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Principal Investigator |
YONEYAMA Akiko University of Tokyo, Department of Medicine, Lecturer, 医学部・附属病院, 講師 (50175684)
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| Co-Investigator(Kenkyū-buntansha) |
AOKI Katsunori University of Tokyo, Department of Medicine, Assistant, 医学部・附属病院, 助手 (40291322)
NAKAHARA Kazuhiko University of Tokyo, Department of Medicine, Professor, 医学部・附属病院, 教授 (70101095)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | flow cytometry / cytokine / 細胞内酵素 / 細胞内サイトカイン |
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| Research Abstract |
Using three-color flow cytometry, intracellular cytokines and surface antigens of T cells were determined simultaneously. Peripheral blood was collected into tubes containing EDTA and was washed twice in RPNfl 1640 supplemented with 20% PCS. It was cultured with Ionomycin (1μg/ml final concentration), PMA (25ng/ml final concentration) and secretion inhibitor Brefeldin A (10μg/ml final concentration) at 37℃ for 4 hours. After permeabilization, IFN-γ and IL-4 accumulated in lymphocytes were stained by FITC or PE- labeled monoclonal antibodies. Cell surface antigens were stained by PerCP-labelled monoclonal antibodies. Since red blood cells of activated samples were difficult to be lysed by usual lysing procedures, lysing procedures were repeated twice. Thus peripheral blood samples collected into tubes containing EDTA could be analyzed for percentages of Th1, Th2, Tc1 and Tc2 lymphocytes, and the results were in good correlation with those obtained for heparinized blood.
Peripheral blood lymphocytes from patients with myelodysplastic syndrome (MDS) and infectious mononucleosis (IM) were analyzed. Peripheral blood lymphocytes from patients with MDS did not show reduced production of cytokines. Most of the CD8^+ lymphocytes which show marked expansion during IM produced IFN-γ and the percentages of Th1 lymphocytes also increased.
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