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

基本信息

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

项目摘要

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.

中性粒细胞功能低下被认为是家畜免疫缺陷综合症的原因。由于中性粒细胞的功能与其产生活性氧的能力密切相关，因此采用电子自旋共振光谱（ESR）和自旋捕获法（ST）相结合的方法来检测和鉴定中性粒细胞中的活性氧。根据所获得的结果，判断该EST-ST方法是否是作为诊断技术的适当方法。中性粒细胞是从牛、马和人的血液中收集的。这些嗜中性粒细胞受到佛波醇肉豆蔻酸酯乙酸酯的刺激。通过ESR-ST方法检测由该刺激产生的活性氧。 2,2,5,5-二甲基吡咯啉N-氧化物(DMPO)用作自旋捕获试剂。化学发光光谱法也被用作检测活性氧产生的另一种方法。当通过ESR-ST方法检查人中性粒细胞产生的活性氧时，证实该细胞产生的活性氧仅为超氧化物（O_2^-）。当将此方法应用于马和牛的中性粒细胞时，观察到马中性粒细胞产生的（O_2^-）量与人类中性粒细胞相同，但牛比人类中性粒细胞少二十分之一和马中性粒细胞。这意味着，虽然 ESR-ST 方法是鉴定所产生的活性氧种类并测量人和马中性粒细胞中活性氧含量的极好方法，但除了 ESR-ST 之外，还使用光学方法来观察化学发光。对于牛的中性粒细胞，必须采用ST法来定量测定(O_2^-)的量。