1999 年度実績報告書

抗がん剤の骨髄抑制に対する可溶性G-CSF受容体の予防効果

研究課題

研究課題/領域番号	11140255
研究機関	九州大学
研究代表者	仁保喜之九州大学, 大学院・医学系研究科, 教授 (60091287)
研究分担者	浅野嘉延九州大学, 医学部, 助手 (60271110) 権藤久司九州大学, 医学部, 助手 (10253428) 岡村孝九州大学, 医学部, 講師 (30136436) 下田和哉九州大学, 医学部, 助手 (90311844)
キーワード	可溶性G-CSF受容体 / G-CSF / 造血前駆細胞 / 抗がん剤 / 骨髄抑制 / CHO細胞
研究概要	抗がん剤治療において骨髄抑制の副作用は必発であり、白血球減少の程度が抗がん剤の投与可能量を規定する。この抗がん剤による骨髄抑制は旺盛な増殖期にある造血前駆細胞が抗がん剤により障害を受けやすいことに起因する。そこで、可溶性G-CSF受容体を用いて内因性G-CSFの作用を阻害し、造血前駆細胞が白血球へ分化増殖する過程を抑制し、一時的に静止期に停めることで抗がん剤による白血球減少を予防する方法に思い至った。この予防法の確立に向けた基礎的検討として、これまでに我々は、CD34陽性細胞(未熟な造血前駆細胞)の約30%にG-CSF受容体の発現を認め、この細胞集団は高率に好中球へと分化すること、可溶性G-CSF受容体がG-CSFの活性を競合的に阻害することなどを明らかにしてきた。そこで、本年度は可溶性G-CSF受容体の大量作成を試みるとともに、可溶性G-CSF受容体の安全性を確認するため生理的な動態を検索した。最初に、cDNAの構造から推定されるG-CSF受容体細胞外部分のペプチドを合成し、それを抗原として家兎で抗体を作成した。この抗体を用いてウエスタンブロットを行なったところ、正常ヒト血清中に80-kDaと85-kDaの2種類の陽性バンドを検出した。この可溶性G-CSF受容体がG-CSFとの結合能を有することも証明した。続いて、G-CSF投与前後の血清と再生不良性貧血の患者の結成を用いて、可溶性G-CSF受容体の血清レベルが好中球に相関することを示した。さらに、G-CSF受容体cDNAの膜貫通部を挟むプライマーを作成しRT-PCRを行ったところ、骨髄細胞が成熟好中球へ分化するに従い可溶性G-CSF受容体の遺伝子発現が高率になることが判明した。これらの結果は、可溶性G-CSF受容体が生体内で好中球造血の調節機構に深く関与している可能性を示唆している。一方、CHO細胞に可溶性G-CSF受容体のcDNAを導入し、培養上清中にG-CSFの作用を阻害する活性を認めた。現在、純化精製による可溶性G-CSF受容体の大量作成を試みている。今後は作成した可溶性G-CSF受容体を用いて、マウスの抗がん剤による骨髄抑制の予防効果を検討する予定である。この結果、抗がん剤による骨髄抑制の予防法が確立できれば、より安全で有効な抗がん剤治療が可能になると期待される。

研究成果

(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Niho Y.: "Molecular Biology of Hematological Disorders"Internal Medicine. 38. 77-78 (1999)
[文献書誌] Iwasaki H.: "Production of soluble granulocyte colony-stimulating factor receptors from myelomonocytic cells"J Immunol. 163. 6907-6911 (1999)
[文献書誌] Mizuo S.: "Interferon-γ prevents apopotosis Epstein-Barr virus-infected natural killer cell leukemia in an autocrine fashion"Blood. 93. 3494-3504 (1999)
[文献書誌] Asano Y.: "Effect of interleukin 10 on the hematopoietic progenitor cells from patients with aplastic anemia"Stem Cells. 17. 147-151 (1999)
[文献書誌] Uchida N.: "Differential gene expression of human telomerase-associated protein hTERT and TEP1 in human hematopoietic cells"Leuk Res. 23. 1127-1132 (1999)
[文献書誌] Okamura K.: "Identification of a novel bovine serum protein which is involved in human T-cell leukemia virus typeI(HTLV-I)induced syncytium formation."Arch Virol. 144. 1-18 (1999)
[文献書誌] Nakashima H.: "The combination of polymorphisms within interderon-γ receptor1(IFN-γ) and receptor2(IFN-γR2) associated with risk of systemic lupus erythematosus(SLE)"FEBS letters. 453. 187-190 (1999)
[文献書誌] Akahoshi M.: "Th1/Th2 balance of the peripheral helper T(Th)cells in systemic lupus erythematosus"Arthritis Rheumatism. 42. 1644-1648 (1999)
[文献書誌] Gao PS.: "Nonpathogenic common variants of IFNγR1 and IFNγR2 in association with total serum IgE levels"Biochem Biophys Res Commun. 263. 425-429 (1999)
[文献書誌] Nakashima H.: "Two polymorphisms within interleukin-3(hIL3)gene detected by mismatch PCR/RFLP"Gene and Immunity. 1. 156-158 (1999)
[文献書誌] Nakashima H.: "Polymorphisms within the II-10 receptor cDNA gene(IL-10R) in Japanese patients with systemic lupus erythematosus(SLE)"Rheumatology. 38. 1142-1144 (1999)
[文献書誌] Nemoto Y.: "Differential effects of interleukin-4 and interleukin-10 on nitric oxide production by murine macrophages"Inflammation Res.. 48. 643-650 (1999)
[文献書誌] Masumoto N.: "V-src induces cisplatin resistance by increasing the repair of cisplatin-DNA interstrand crosslinks in human gallbladder adenocarcinoma cells"Int. J. Cancer. 80. 731-737 (1999)
[文献書誌] Nagata C.: "Hair dye use and occupational exposure to organic solvents as risk factors for myelodysplastic syndrome"Leukemia Res.. 23. 57-62 (1999)
[文献書誌] Okuma K.: "Host range of human T-cell leukemia virus type I analyzed by a cell fusion-dependent reporter gene activation assay"Virology. 254. 235-244 (1999)
[文献書誌] Tanaka Y.: "Association of the interferon-γ receptor variant (Vall4Met)with systemic lupus erythematosus"Immunogenetics. 49. 266-271 (1999)
[文献書誌] Takasaki S.: "Elevated serum soluble CD8 level in aoutoimmune hepatities and the effect of corticsteroid therapy"Hepatology Res.. 15. 52-63 (1999)
[文献書誌] Horiuhi T.: "An NcoI polymorphism in the human complement component 7(C7)gene"J. Hum. Genet.. 44. 270-271 (1999)
[文献書誌] Nagafuchi S.: "Establishment of an embrvonic stem(ES)cell line derived frome a non-obese deiabetic (NOD)mouse : in vivo differentiation into lymphocytes and potential for germ line transmission"FEBS Letters. 455. 101-104 (1999)
[文献書誌] Yamaoka K.: "Selective DNA-binding activity of interleukin-10-stimulated STAT molecules in human monocytes"J. Interferon and Cytokine Res.. 19. 679-685 (1999)
[文献書誌] Higuchi M.: "Establishment and characterization of a Fas-Resistant T cell line"Acta Haematol.. 102. 22-30 (1999)
[文献書誌] Togawa A.: "Progressive impairment of kidneys and reproductive organs in mice lacking Pho GDI α"Oncogene. 18. 5373-5380 (1999)
[文献書誌] Nakamura T.: "A clonicopathological study in young patients with gastric carcinoma"J. Surgical Oncology. 71. 214-219 (1999)
[文献書誌] Arinobu Y.: "Antagonistic effects of an alternative splice variant of human IL-4, II-4δ2, on IL-4 activities in human monocytes and B cells"Cell. Immunol.. 191. 161-167 (1999)

1999 年度 実績報告書

抗がん剤の骨髄抑制に対する可溶性G-CSF受容体の予防効果

研究代表者

仁保 喜之 九州大学, 大学院・医学系研究科, 教授 (60091287)

研究成果

[文献書誌] Niho Y.: "Molecular Biology of Hematological Disorders"Internal Medicine. 38. 77-78 (1999)

[文献書誌] Iwasaki H.: "Production of soluble granulocyte colony-stimulating factor receptors from myelomonocytic cells"J Immunol. 163. 6907-6911 (1999)

[文献書誌] Mizuo S.: "Interferon-γ prevents apopotosis Epstein-Barr virus-infected natural killer cell leukemia in an autocrine fashion"Blood. 93. 3494-3504 (1999)

[文献書誌] Asano Y.: "Effect of interleukin 10 on the hematopoietic progenitor cells from patients with aplastic anemia"Stem Cells. 17. 147-151 (1999)

[文献書誌] Uchida N.: "Differential gene expression of human telomerase-associated protein hTERT and TEP1 in human hematopoietic cells"Leuk Res. 23. 1127-1132 (1999)

[文献書誌] Okamura K.: "Identification of a novel bovine serum protein which is involved in human T-cell leukemia virus typeI(HTLV-I)induced syncytium formation."Arch Virol. 144. 1-18 (1999)

[文献書誌] Nakashima H.: "The combination of polymorphisms within interderon-γ receptor1(IFN-γ) and receptor2(IFN-γR2) associated with risk of systemic lupus erythematosus(SLE)"FEBS letters. 453. 187-190 (1999)

[文献書誌] Akahoshi M.: "Th1/Th2 balance of the peripheral helper T(Th)cells in systemic lupus erythematosus"Arthritis Rheumatism. 42. 1644-1648 (1999)

[文献書誌] Gao PS.: "Nonpathogenic common variants of IFNγR1 and IFNγR2 in association with total serum IgE levels"Biochem Biophys Res Commun. 263. 425-429 (1999)

[文献書誌] Nakashima H.: "Two polymorphisms within interleukin-3(hIL3)gene detected by mismatch PCR/RFLP"Gene and Immunity. 1. 156-158 (1999)

[文献書誌] Nakashima H.: "Polymorphisms within the II-10 receptor cDNA gene(IL-10R) in Japanese patients with systemic lupus erythematosus(SLE)"Rheumatology. 38. 1142-1144 (1999)

[文献書誌] Nemoto Y.: "Differential effects of interleukin-4 and interleukin-10 on nitric oxide production by murine macrophages"Inflammation Res.. 48. 643-650 (1999)

[文献書誌] Masumoto N.: "V-src induces cisplatin resistance by increasing the repair of cisplatin-DNA interstrand crosslinks in human gallbladder adenocarcinoma cells"Int. J. Cancer. 80. 731-737 (1999)

[文献書誌] Nagata C.: "Hair dye use and occupational exposure to organic solvents as risk factors for myelodysplastic syndrome"Leukemia Res.. 23. 57-62 (1999)

[文献書誌] Okuma K.: "Host range of human T-cell leukemia virus type I analyzed by a cell fusion-dependent reporter gene activation assay"Virology. 254. 235-244 (1999)

[文献書誌] Tanaka Y.: "Association of the interferon-γ receptor variant (Vall4Met)with systemic lupus erythematosus"Immunogenetics. 49. 266-271 (1999)

[文献書誌] Takasaki S.: "Elevated serum soluble CD8 level in aoutoimmune hepatities and the effect of corticsteroid therapy"Hepatology Res.. 15. 52-63 (1999)

[文献書誌] Horiuhi T.: "An NcoI polymorphism in the human complement component 7(C7)gene"J. Hum. Genet.. 44. 270-271 (1999)

[文献書誌] Nagafuchi S.: "Establishment of an embrvonic stem(ES)cell line derived frome a non-obese deiabetic (NOD)mouse : in vivo differentiation into lymphocytes and potential for germ line transmission"FEBS Letters. 455. 101-104 (1999)

[文献書誌] Yamaoka K.: "Selective DNA-binding activity of interleukin-10-stimulated STAT molecules in human monocytes"J. Interferon and Cytokine Res.. 19. 679-685 (1999)

[文献書誌] Higuchi M.: "Establishment and characterization of a Fas-Resistant T cell line"Acta Haematol.. 102. 22-30 (1999)

[文献書誌] Togawa A.: "Progressive impairment of kidneys and reproductive organs in mice lacking Pho GDI α"Oncogene. 18. 5373-5380 (1999)

[文献書誌] Nakamura T.: "A clonicopathological study in young patients with gastric carcinoma"J. Surgical Oncology. 71. 214-219 (1999)

[文献書誌] Arinobu Y.: "Antagonistic effects of an alternative splice variant of human IL-4, II-4δ2, on IL-4 activities in human monocytes and B cells"Cell. Immunol.. 191. 161-167 (1999)

1999 年度実績報告書

仁保喜之九州大学, 大学院・医学系研究科, 教授 (60091287)