| Project/Area Number |
14360180
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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 |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Osaka Prefecture University |
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Principal Investigator |
SUGIURA Kikuya Osaka Prefecture University, Graduated School of Life and Environmental Sciences, Associated Professor, 生命環境科学研究科, 助教授 (30171143)
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| Co-Investigator(Kenkyū-buntansha) |
INABA Toshio Osaka Prefecture University, Graduated School of Life and Environmental Sciences, Professor, 生命環境科学研究科, 教授 (00137241)
KUMAGAI Daijiro Osaka Prefecture University, Graduated School of Life and Environmental Sciences, Research Associate, 生命環境科学研究科, 助手 (70316016)
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| Project Period (FY) |
2002 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | Dog / Bone marrow transplantation / Marrow collection methods / Pre-transplantation conditioning / Hematopoietic stem cells / Isolation / Monoclonal antibody / Immunosuppressant / 造血前駆細胞 / Tリンパ球 / 移植片対宿主反応 / 免疫抑制 / 多能性造血コロニー形成細胞 / 小麦胚芽凝集素(WGA) / Rhodamine123 / 比重分画 / セルソーティング / 低比重細胞分画 / 造血コロニー形成細胞 / 骨髄破壊性前処置法 / 骨髄非破壊性前処置法 / ドナーリンパ球輸注 / マイクロサテライト / フローサイトメートリー / コンディショニング |
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| Research Abstract |
1.Establishment of methods for marrow collection, pre-treatment of recipients and promotion of engraftment in dog allogeneic bone marrow transplantation (BMT) :
1)Marrow collection method : Perfusion collection method and aspiration collection method were compared. Bone marrow (BM) cells collected by perfusion method heve more hematopoietic progenitor cells detected with canine CD34, and significantly less red blood cell count
2)Pre-transplantation conditioning of recipients : Engraftment of donor bone marrow cells was found by pre-treating recipients with some combinations of myeloablative and nonmyeloablative conditioning methods preceding BMT
3)Promotion of BMC engraftment : We did donor- lymphocyte- infusion (DLI) for promotion of BMC engraftment, and found that white blood cell counts significantly elevated after DLI.
2.Isolation and characterization of canine hematopoietic stem cells : We fractionated BMCs by cell density, affinity of wheat germ agglutinin (WGA), and stainability of Rhodamine123 (Rh), and found that most of multipotent hematopoietic progenitor cells (MHPCs) in BM were contained in the fraction of cells showing low density OLD), high affinity of WGA (WGA^<high>), and low stainability of Rh (Rh^<low>).
3.Establishment of monoclonal antibodies (mAbs) to recognize MHPCs : We immunized WGA^<high> LD BMCs of dog to mice, and obtained several clones of mAb which recognize surface molecule on MHPCs.
4.Establishment of mAb to canine T cells : We established a mAb against canine T cells, which suppresses immune responses in the mixed lymphocyte cultures and decrease T cell counts in peripheral blood after intravenous injection.
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