The mechanisms of atheroma formation in vessel wall using a unique monoclonal antibody(ASH1a/256C) recognizing. atherosclerotic lesions.

使用独特的单克隆抗体（ASH1a/256C）识别血管壁粥样斑块形成的机制。

• 批准号: 14572068
• 负责人: MORI Masahiro
• 金额: $ 1.92万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572068/
• 关键词: atherosclerosis; foam cell; cholesterol; apoptosis; lipid droplets; modified LDL; monoclonal antibody; non-invasive diagnosis; マクロファージ; Cholesterol; Phosphatidylcholine; 非浸襲的診断

In order to investigate application for low invasive diagnosis of cardiovascular disease, novel monoclonal antibodies (ASH1a/256C) recognizing fatty streaks of atheroma in vivo and in vitro were prepared. The ASH1a/256C antigen purified from WHHL rabbit (a model animal for familial hyper The expression of PC-cholesterol complex correlates foam cell rupture and accumulation of lipid in extracellular space in intima) atheroma was determined as "PC-cholesterol complex" specifically localized in extracellular space in thickened intima.Foam cells can produce this antigen as "FC (free cholesterol) enriched lipid droplets", only when macrophages incubated with Acetylated LDL, WHHL rabbit serum or Human hyperlipidemic serum. VLDL/IDL could also induce foam cell formation, however, no FC-rich lipid droplets were observed. The foam cells containing FC-rich lipid droplets were cultured continuously until 2 weeks ; finally, the foam cells ruptured and lipid droplets remained in extracellular matrix.When the macrophage incubated with cholesterol oleate liquid crystals (not contain PC), macrophages could also produce PC-cholesterol complex (FC-rich lipid droplets) and change to foam cells morphologically, Since U18666A inhibited cellular cholesterol transport, when the foam cell incubated continuously with this material, foam cells failed to rupture themselves. These observation suggest that hydrolysis of CE (cholesteryl ester) and cellular cholesterol transport are important process for the formation of FC-rich lipid dropletsThe expression of PC-cholesterol complex correlates foam cell rupture and accumulation of lipid in extracellular space in intima

为了调查对心血管疾病的低浸润性诊断的应用，制备了新型的单克隆抗体（ASH1A/256C），以识别体内和体外的动脉粥样硬化的脂肪条纹。 The ASH1a/256C antigen purified from WHHL rabbit (a model animal for familial hyper The expression of PC-cholesterol complex correlates foam cell rupture and accumulation of lipid in extracellular space in intima) atheroma was determined as "PC-cholesterol complex" specifically localized in extracellular space in thickened intima.Foam cells can produce this antigen as "FC (free cholesterol)富集的脂质液滴”，只有当巨噬细胞与乙酰化的LDL，WHHL兔血清或人类高脂血清孵育时。 VLDL/IDL也可能诱导泡沫细胞的形成，但是，未观察到富含FC的脂质液滴。连续培养含有富含FC的脂质液滴的泡沫细胞直至2周。最后，泡沫细胞破裂，脂质液滴保留在细胞外基质中。当与胆固醇的巨噬细胞孵育时，胆固醇液晶（不包含PC），巨噬细胞还会产生PC-胆固醇复合物（富含FC的脂质液滴），并在泡沫中转移到泡沫状细胞上，自从使用U18666666a时，泡沫细胞与这种材料不断孵育，无法自行破裂。这些观察结果表明，CE（胆固醇酯）和细胞胆固醇转运的水解是形成FC富脂质液滴的重要过程

项目成果

Itabe, H., Mori M., Fujimoto, Y., Higashi, Y., Takano, T.: "Minimally modified LDL is an oxidized LDL enriched with oxidized phosphatidylcholine"J.Biochem(Tokyo). 134. 459-465 (2003)

Itabe, H.、Mori M.、Fujimoto, Y.、Higashi, Y.、Takano, T.：“最小修饰的 LDL 是富含氧化磷脂酰胆碱的氧化 LDL”J.Biochem（东京）。

Fujumoto, Y., Itabe, H., Mori, M., Higashi, Y., Kojima, S.Takano, T.: "Identification of the major lipid, droplet proteins of the human hepatocyte cell line HmH7"Biochemica. Biophysica. Acta. 1644. 47-59 (2003)

Fujumoto, Y.、Itabe, H.、Mori, M.、Higashi, Y.、Kojima, S.Takano, T.：“人肝细胞系 HmH7 的主要脂质、液滴蛋白的鉴定”Biochemica。

Higashi, Y., Itabe, H., Fujimoto, Y., Takano T.: "Distribution of Microsomal triglyceride transfer protein within sub-endoplasmic reticulum regions in human hepatoma cells"Biochemica. Biophysica. Acta. 1581. 127-136 (2002)

Higashi, Y.、Itabe, H.、Fujimoto, Y.、Takano T.：“微粒体甘油三酯转移蛋白在人肝癌细胞亚内质网区域内的分布”Biochemica。

Itabe, H., Mori, M., Fujimoto, Y., Higashi, Y., Takano, T.: "Minimally modified LDL is an oxidized LDL enriched with oxidized phosphatidyicholine"J. Biochem. (Tokyo). 134. 459-465 (2003)

Itabe, H.、Mori, M.、Fujimoto, Y.、Higashi, Y.、Takano, T.：“最小修饰的 LDL 是一种富含氧化磷脂酰胆碱的氧化 LDL”。

Higashi, Y., Itabe, H., Fukase, H., Mori, M., Fujimoto, Y., Sato, R., Imanaka, T, Takano, T.: "Distribution of microsomal triglyceride transfer protein within sub-endoplasmic reticulum regions in human hepatoma cells"Biochemica et Biophysica Acta. 1581. 1

Higashi, Y.、Itabe, H.、Fukase, H.、Mori, M.、Fujimoto, Y.、Sato, R.、Imanaka, T、Takano, T.：“微粒体甘油三酯转移蛋白在亚内质中的分布