Generation and Role of Active oxygens in Cells

细胞中活性氧的产生和作用

• 批准号: 05044180
• 负责人: NAKAMURA Michio
• 金额: $ 2.56万
• 依托单位: Institute of Tropical Medicine, Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05044180/
• 关键词: Superoxide-Generating System; Phagocytes; B Lymphocytes; Chronic granulomatoous disease; Cytochrome b558; 活性酸素産生

1. For characterizing superoxide-generating system of phagocytes, we analyzed the cells of the patients with chronic granulomatous disease (CGD), because their genetic backgrounds have apparently well understood. The system is composed of 4 components, namely gp91-phox, p22-phox, p47-phox, and p67-phox. The former two make a cytochrome b558 lying on the membrane of phagocytes. In a patient, a common genetic phenotype has been noted to prevail over all kinds of his phagocytes. We, however, have discovered a 62-year male patient whose granulocyte fraction definitely contain a small population with normal amount of surface cytochrome b558. The population has been identified as eosinophils. Neither gp91-phox nor phorbol ester-stimulated chemiluminescence (CL) response are observed in his neutrophils and monocytes. His eosinophils definitely contain gp91-phox and exhibit the normal extent of CL response. Molecular analyzes have shown that coding sequences of his gp91-phox mRNA and its gene, and the poly (A) signal of the gene are normal. We have discovered a point mutation in the 5'-flanking region of his gp91-phox gene and are now extensively characterizing the function of that mutated region.2. For characterizing superoxide-generating system of non-phagocytic cells, we used B lymphocytes, fibroblasts, mesangial cells, and others. The system of B lymphocytes is essential' the same as that of phagocytes. The expression of gp91-phox in B lymphocytes should be the same as that of neutrophils and monocytes but not of eosinophils. The amount of cytochrome b558 in B lymphocytes is quite low comparing to that of granulocytes, which corresponds to low CL response of the lymphocytes. Fibroblasts may have a different cytochrome because the cells of the patient with CGD normally generate superoxide anions. The superoxide-generating system of non-phagocytic cells should be hereafter reexamined by using molecular probes prepared for phagocytic cells.

1.为了表征吞噬细胞的超氧化物生成系统，我们分析了慢性肉芽肿病（CGD）患者的细胞，因为他们的遗传背景显然已经很好地了解。该系统由4个组分组成，即gp 91-phox、p22-phox、p47-phox和p67-phox。前两者使细胞色素b558位于吞噬细胞膜上。在一个病人，一个共同的遗传表型已被注意到占主导地位的各种他的吞噬细胞。然而，我们发现了一个62岁的男性患者，其粒细胞部分肯定含有少量细胞色素b558的表面量正常。该群体已被确定为嗜酸性粒细胞。在他的中性粒细胞和单核细胞中既没有观察到gp 91-phox也没有观察到佛波酯刺激的化学发光（CL）反应。他的嗜酸性粒细胞肯定含有gp 91-phox，并表现出正常程度的CL反应。分子生物学分析表明，hisgp 91-phox mRNA及其基因的编码序列和该基因的poly（A）信号正常。我们已经在他的gp 91-phox基因的5 '侧翼区域发现了一个点突变，现在正在广泛地表征该突变区域的功能。为了表征非吞噬细胞的超氧化物生成系统，我们使用了B淋巴细胞、成纤维细胞、系膜细胞等。B淋巴细胞的系统与吞噬细胞的系统是基本相同的。gp 91-phox在B淋巴细胞中的表达应与中性粒细胞和单核细胞相同，而不是嗜酸性粒细胞。与粒细胞相比，B淋巴细胞中细胞色素b558的量非常低，这对应于淋巴细胞的低CL反应。成纤维细胞可能具有不同的细胞色素，因为CGD患者的细胞通常产生超氧阴离子。此后应使用为吞噬细胞制备的分子探针重新检查非吞噬细胞的超氧化物生成系统。

项目成果

Kumatori, Atsushi: "A patient with "variant' chronic granulomatous disease with normal eosinophils" Jap J Bacteriol. 50 (1). 235-235 (1995)

Kumatori, Atsushi：“一名患有‘变异’慢性肉芽肿病且嗜酸性粒细胞正常的患者”Jap J Bacteriol. 50 (1). 235-235 (1995)

J.H.W.Leusen,M.de Boer,B.G.J.M.Bolscher,P.H.Hilarius,R.S.Weening,H.D.Ochs,D.Roos,A.J.J.Verhoeven: "A pointmutation in gp91-phox of cytochrome b558 of the human NADPH oxidase leading to defective translocation of the cytosolic proteins p47-phox and p67-pho

J.H.W.Leusen、M.de Boer、B.G.J.M.Bolscher、P.H.Hilarius、R.S.Weening、H.D.Ochs、D.Roos、A.J.J.Verhoeven：“人 NADPH 氧化酶细胞色素 b558 的 gp91-phox 中的点突变导致胞浆蛋白缺陷易位

Nakamura, Michio: Nankodo (Tokyo). "The Role of Phagocytes" in Medical Bacteriology (Ed.Yoshikawa, S), Inpress (1995)

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Kosaka,Mitsuo: ""The role of heat shock proteins and interleukin-1 interaction in suppression of fever"in Temperature Regulation(Ed.Milton,AS)" Birkhauser Verlag(Basel), 5 (1994)

Kosaka，Mitsuo：“温度调节中热休克蛋白和白细胞介素 1 相互作用在抑制发烧中的作用”（Ed.Milton，AS）” Birkhauser Verlag（巴塞尔），5（1994）

熊取,厚志: "新奇な易感染性慢性肉芽腫症" 日本細菌学会雑誌. 50. 235-235 (1995)

Kumatori，Atsushi：“新型易感慢性肉芽肿病”日本细菌学会杂志 50. 235-235 (1995)。