Possibility of Electron Spin Resonance Spectroscopy as a Diagnostic Technique in Immunodeficiency Syndrome of Domestic Animals

电子自旋共振光谱作为家畜免疫缺陷综合症诊断技术的可能性

• 批准号: 01560333
• 负责人: KUWABARA Mikinori
• 金额: $ 0.96万
• 依托单位: Hokkaido 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: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01560333/
• 关键词: Immunodeficiency Syndrome; Granulocyte Hypofunction Syndrome; neutrophil; Superoxide; Electron Spin Resonance Spectroscopy; Spin Trapping Method; NADPH Oxidase; Phorbol 12-Myristate 13-Acetate; ホルボ-ルミリステ-トアセテ-ト; Immunodeficiency Syndrome; Granulocyte Hypofunction Syndrome; neutrophil; Superoxide; Electron Spin Resonance Spectroscopy; Spin Trapping Method; NADPH Oxidase; Phorbol 12-Myristate 13-Acetate; ホルボ-ルミリステ-トアセテ-ト

Hypofunction of neutrophil cells is thought to be a cause of immunodeficiency syndrome of domestic animals. Since a function of neutrophil cells has been proved to be closely related to their ability to generate reactive oxygens, a method combining electron spin resonance spectroscopy (ESR) and spin trapping method (ST) was employed to detect and identify the reactive oxygens from neutrophil cells. On the basis of the results obtained, this EST-ST method was judged to be or not to be an appropriate method as a diagnostic technique. Neutrophil cells was collected from cow, horse and human bloods. These neutrophils were stimulated by phorbol myristate acetate. The reactive oxygens which were generated by this stimulation were detected by the ESR-ST method. 2, 2, 5, 5-Dimethylpyrroline N-oxide (DMPO) was used as a spin trapping reagent. Chemiluminescence spectroscopy was also employed as another method to detect the generation of reactive oxygens.When the generation of reactive oxygens from human neutrophil cells was examined by the ESR-ST method, it was confirmed that the reactive oxygen which was generated from this cell was only superoxide (O_2^-). When this method was applied to the neutrophils of horse and cow, it was observed that the amount of (O_2^-) generated from horse neutrophils was the same as that of human netrophils, but that of cow was a twentieth less than those of human and horse neutrophils. This means that although the ESR-ST method is an excellent method to identify species of the generated reactive oxygens and to measure the amount of them in human and horse neutrophils, but the use of an optical method to observe chemilumineescence in addition to the ESR-ST method was necessary to quantitatively measure the amount of (O_2^-)in the case of neutrophils from cow.

中性粒细胞的功能不全被认为是家畜免疫缺陷综合征的原因。由于中性粒细胞的功能已被证明与它们产生活性氧的能力密切相关，因此采用了一种结合电子自旋谐振光谱（ESR）和自旋陷阱方法（ST）的方法来检测和鉴定来自中性粒细胞的反应性氧。根据获得的结果，这种EST-ST方法被认为是作为诊断技术的适当方法。从牛，马和人类血液中收集中性粒细胞。这些嗜中性粒细胞是通过佛罗其肉豆蔻酸酯乙酸盐刺激的。通过ESR-ST方法检测到该刺激产生的反应性氧。 2、2、5、5-二甲基吡咯氨酸N-氧化物（DMPO）用作自旋诱捕试剂。化学发光光谱也被用作检测反应性氧的产生的另一种方法。当通过ESR-ST方法研究了人类嗜中性粒细胞的活性氧的产生时，可以证实，从该细胞中产生的活性氧仅是超氧化物（O_2^ - ）。当将这种方法应用于马和牛的中性粒细胞时，观察到从马中性粒细胞产生的（O_2^ - ）的量与人类网络粒细胞的含量相同，但牛的含量比人类和中性粒细胞的牛少于20。这意味着，尽管ESR-ST方法是鉴定产生的活性氧的物种并测量人类和马中性粒细胞中它们的数量的一种极好方法，但是除了使用eSR-ST方法观察化学方法外，还必须使用光学方法来定量地测量（O_2^ - ）的量（O_2^ - ）。