Purification of maturation-induced factor of megakaryoblastic cell line

巨核细胞系成熟诱导因子的纯化

• 批准号: 06671105
• 负责人: ISHIDA Yoji
• 金额: $ 1.41万
• 依托单位: IWATE MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671105/
• 关键词: megakaryoblastic leukemia; maturation-induced factor

A humoral factor which induces the maturation of megakaryoblastic cell line was purified from the sources including the supernatant of leukemic cell lines, the urine obtained from the patients with aplastic anemia and rabbit thrombocytopenic plasma. Biological effects such as (1) Achetylcholinesterase (AchE) expression of murine megakaryoblastic cell line L8057, (2) Glycoprotein IIb/IIIa expression of human megakaryoblastic cell line CMK (3) The raito to induce CMK cells to S-phase (4) AchE expression of murine bone marrow cells were studied using this materials. The activity was not found in the supernatant of leukemic cell lines, but in the urine obtained from the patients with aplastic anemia and rabbit thrombocytopenic plasma. Therefore I purified the factor from the urine obtained from the patients with aplastic anemia or rabbit thrombocytopenic plasma based on the AchE expression of murine bone marrow cells. The factor has not been purified because of small amount of volume. The protein was purified from rabbit thrombocytopenic plasma using 6 steps including ammonium sulfate precipitation, WGL sepharose, Blue sepharose, Ni^<2+> sepharose and gel filtration. Interestingly, this protein stimulated proplatelet formation from mature megakaryocytes.

从白血病细胞培养上清、再生障碍性贫血患者尿液和兔血小板减少血浆中分离纯化了一种诱导巨核细胞成熟的体液因子。用该材料研究了（1）小鼠巨核细胞系L 8057乙酰胆碱酯酶（AchE）表达，（2）人巨核细胞系CMK糖蛋白IIb/IIIa表达，（3）CMK细胞S期诱导率，（4）小鼠骨髓细胞AchE表达等生物学效应。白血病细胞培养上清中未发现此活性，但在再生障碍性贫血患者尿液和兔血小板减少血浆中发现此活性。因此，根据小鼠骨髓细胞AchE的表达，我从再生障碍性贫血患者的尿液或兔血小板减少的血浆中纯化了该因子。该因子由于体积小，未进行纯化。采用硫酸铵沉淀、WGL琼脂糖凝胶、Blue琼脂糖凝胶、Ni^2+琼脂糖凝胶和凝胶过滤6步纯化兔血小板减少血浆中的蛋白。有趣的是，这种蛋白质刺激成熟巨核细胞的前血小板形成。

项目成果

Takaaki Beppu: "Detection of S phase cells with an antibody to proliferating cell nuclear antigen (PCNA)." J. Histotechnology. 17. 325-328 (1994)

Takaaki Beppu：“用增殖细胞核抗原 (PCNA) 抗体检测 S 期细胞。”

Sasaki Kohsuke: "Estimation of S-phase fraction in tumor tissue sections by immunohistochemical staining of PCNA" J.Histochem Cytochem. 42. 957-960 (1994)

Sasaki Kohsuke：“通过 PCNA 免疫组织化学染色估计肿瘤组织切片中的 S 期分数”J.Histochem Cytochem。

Yoji Ishida: "Thrombopoietin" Clinical Immunology. 27. 240-249 (1995)

石田洋二：“血小板生成素”临床免疫学。

Yoji Ishida: "Thrombopoietin" Hematology/Oncology. 31. 381-391 (1995)

Yoji Ishida：“血小板生成素”血液学/肿瘤学。

Beppu Takaaki: "Identification of S-phase cells using PC10 antibody to proliferocting cell nuclear antigen(PCNA)in flow cytometric analysis" J.Histochem Cytochem. 42. 1177-1182 (1994)

Beppu Takaaki：“在流式细胞术分析中使用抗增殖细胞核抗原 (PCNA) 的 PC10 抗体鉴定 S 期细胞”J.Histochem Cytochem。