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

中性粒细胞功能低下被认为是家畜免疫缺陷综合征的原因之一。由于中性粒细胞的功能已被证明与其产生反应性氧的能力密切相关，因此采用电子自旋共振光谱(ESR)和自旋捕捉法(ST)相结合的方法来检测和鉴定中性粒细胞中的反应性氧。根据所获得的结果，判断该EST-ST方法作为一种诊断技术是否合适。从牛、马和人的血液中采集中性粒细胞。这些中性粒细胞被佛波醇肉豆蔻酸醋酸酯刺激。用ESR-ST法检测刺激产生的反应性氧。用2，2，5，5-二甲基吡咯烷N-氧化物(DMPO)作为自旋捕捉剂。化学发光光谱也是检测活性氧产生的另一种方法，当用ESR-ST法检测人中性粒细胞产生活性氧时，证实该细胞产生的活性氧只是超氧化物(O_2^-)。将这种方法应用于马和牛的中性粒细胞，观察到马的中性粒细胞产生的(O_2^-)量与人的中性粒细胞相同，但牛的中性粒细胞产生的(O_2^-)量比人和马的少二十分之一。这意味着，虽然ESR-ST方法是鉴定人和马中性粒细胞中产生的反应性氧的种类和数量的一种很好的方法，但对于奶牛中性粒细胞，除了ESR-ST方法外，还需要用光学方法观察化学发光来定量测量(O_2^-)的数量。

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.