In vitro expansion of human and mouse mast cells by the ligand for c-ki

c-ki 配体体外扩增人和小鼠肥大细胞

• 批准号: 03454262
• 负责人: NAKAHATA Tatsutoshi
• 金额: $ 3.78万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454262/
• 关键词: mast cells; SCF; IL-3; mouse; human; hemopoietic stem cell; CTMC; MMC; stem cell factor (SCF); cーkit リガンド; ILー3; ILー4; 肥満細胞コロニ-; 大量培養

Mast cell are derived from multipotential hematopoietic stem cells and classified into at least two phenotypically distinct subpopulations; connective tissue-type mast cells (CTMC) and mucosal mast cells (MMC). When normal mouse bone marrow cells were cultured with IL-3, a large number of mast cells ,which were tissue culture equivalents of the MMC subclass, were developed. Mature CTMC purified from mouse peritoneal calls could proliferate in the presence of both IL-3 and IL-4. Stem cell factor (SCF), the ligand for the c-kit tyrosine kinase receptor, demonstrated to act synergistically with a variety of cytokines including IL-3, IL-6, and G-CSF on the development of early hematopoietic progenitors. Although SCF alone failed to induce mast cells from bone marrow lineage negative cells, it stimulated mast cell production in combination with IL-3. Either SCF, IL-3 alone could not induce mast cell colonies from purified mature CTMC, but SCF revealed significant cooperative actions with IL-3, IL-4 or GM-CSF on the proliferation of mouse CTMC in vitro. The anti-c-kit receptor antibody (ACK2) completely blocked the activity of SCF but not of IL-3 on the growth of CTMC. When we cultured human CD34^+ cells purified from cord blood or bone marrow cells in the presence of SCF, human mast cells first detected on 14 days of incubation and increased dramatically after 50 days of culture. More than 90% of the cells in culture with SCF,SCF+IL-6 or SCF+IL-11 were mast cells after 60 days of culture of cord blood CD34^+ cells. IL-3 and GM-CSF significantly inhibited SCF-dependent mast cell proliferation.

肥大细胞源自多能造血干细胞，并分为至少两个表型不同的亚群；结缔组织型肥大细胞（CTMC）和粘膜肥大细胞（MMC）。当正常小鼠骨髓细胞与 IL-3 一起培养时，会产生大量肥大细胞，这些肥大细胞相当于 MMC 亚类的组织培养物。从小鼠腹膜细胞中纯化的成熟 CTMC 在 IL-3 和 IL-4 存在的情况下可以增殖。干细胞因子 (SCF) 是 c-kit 酪氨酸激酶受体的配体，已被证明与多种细胞因子（包括 IL-3、IL-6 和 G-CSF）对早期造血祖细胞的发育具有协同作用。虽然单独使用 SCF 无法从骨髓谱系阴性细胞中诱导肥大细胞，但它与 IL-3 结合可刺激肥大细胞产生。单独使用SCF、IL-3均不能从纯化的成熟CTMC中诱导肥大细胞集落，但SCF与IL-3、IL-4或GM-CSF在体外对小鼠CTMC增殖具有显着的协同作用。抗c-kit受体抗体(ACK2)完全阻断SCF但不阻断IL-3对CTMC生长的活性。当我们在 SCF 存在下培养从脐带血或骨髓细胞纯化的人类 CD34^+ 细胞时，人类肥大细胞在培养 14 天时首次检测到，并在培养 50 天后急剧增加。脐带血CD34^+细胞培养60天后，SCF、SCF+IL-6或SCF+IL-11培养物中90%以上的细胞为肥大细胞。 IL-3 和 GM-CSF 显着抑制 SCF 依赖性肥大细胞增殖。

项目成果

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中村 友彦,松岡 高史,馬場 淳,田中 昌明,津野 隆久,小池 健一,中畑 龍俊,小宮山 淳: "未熟児貧血に対するリコンビナント・ヒト・エリスロポエチンの臨床効果." 日児誌. 95. 1105-1109 (1991)

Tomohiko Nakamura、Takashi Matsuoka、Jun Baba、Masaaki Tanaka、Takahisa Tsuno、Kenichi Koike、Tatsutoshi Nakahata、Jun Komiyama：“重组人促红细胞生成素对早产儿贫血的临床效果”95。1105-1109（1991）。

Seki T.,Nakahata T.,et al.: "A case of cell mediated immune-pancytopenia complicating primary Sjogren's syndrome." Am.J.Hematol.

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