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鲁米诺和光泽精增强火鸡单核细胞的化学发光

Lucigenin- and luminol-enhanced chemiluminescence in turkey monocytes.

作者信息

Van Nerom A, Desmidt M, Ducatelle R, Haesebrouck F

机构信息

Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, University of Ghent, Belgium.

出版信息

J Biolumin Chemilumin. 1997 Jul-Aug;12(4):207-14. doi: 10.1002/(SICI)1099-1271(199707/08)12:4<207::AID-BIO446>3.0.CO;2-2.

Abstract

Monocytes from 10 week-old specific pathogen-free turkeys were isolated from peripheral blood by density centrifugation and assayed for their oxidative activity by means of a luminometer. Chemiluminescence (CL) properties after stimulation with different soluble and particulate stimuli were compared in lucigenin- and luminol-enhanced assays. A distinct response could be measured with 12-phorbol 13-myristate acetate (PMA) and Zymosan A, but only a weak signal was obtained with calcium ionophore A23187. No oxidative activity could be induced with N-formyl-methionyl-phenylalanine. Peak maxima for both lucigenin- and luminol-enhanced CL were ranked: PMA > Zymosan A > calcium ionophore. The velocity of the lucigenin- and luminol-enhanced responses induced by calcium ionophore were of similar magnitude, but the lucigenin-enhanced responses of Zymosan A and PMA-stimulated monocytes were respectively about 5 and 10 times higher than those obtained in luminol-enhanced assays. No peroxidase activity could be detected in the purified turkey monocytes. As luminol-enhanced CL primarily results from the peroxidase activity, this lack of myeloperoxidase may explain the observed lower responses to the different stimuli, in the presence of a luminol. In contrast, lucigenin-enhanced CL is not related to peroxidase activity, but is a selective probe of oxidase activity. Irrespective of the myeloperoxidase deficiency, different soluble and particulate stimuli induced a significant and reproducible CL response in turkey monocytes, in the presence of both chemiluminigenic probes, lucigenin and luminol. The possibility of measuring the phagocyte oxygenation activity of turkey monocytes represents a useful tool for the study of monocyte mediated host defence in the turkey.

摘要

通过密度离心法从10周龄无特定病原体火鸡的外周血中分离出单核细胞,并用发光计检测其氧化活性。在光泽精和鲁米诺增强的试验中,比较了用不同可溶性和颗粒性刺激物刺激后的化学发光(CL)特性。用12 - 佛波醇13 - 肉豆蔻酸酯乙酸酯(PMA)和酵母聚糖A可检测到明显的反应,但用钙离子载体A23187仅获得微弱信号。N - 甲酰 - 甲硫氨酰 - 苯丙氨酸不能诱导氧化活性。光泽精和鲁米诺增强的CL的峰值最大值排序为:PMA>酵母聚糖A>钙离子载体。钙离子载体诱导的光泽精和鲁米诺增强反应的速度大小相似,但酵母聚糖A和PMA刺激的单核细胞的光泽精增强反应分别比鲁米诺增强试验中获得的反应高约5倍和10倍。在纯化的火鸡单核细胞中未检测到过氧化物酶活性。由于鲁米诺增强的CL主要源于过氧化物酶活性,这种髓过氧化物酶的缺乏可能解释了在存在鲁米诺的情况下对不同刺激观察到的较低反应。相比之下,光泽精增强的CL与过氧化物酶活性无关,而是氧化酶活性的选择性探针。无论髓过氧化物酶缺乏情况如何,在存在化学发光探针光泽精和鲁米诺的情况下,不同可溶性和颗粒性刺激物在火鸡单核细胞中诱导了显著且可重复的CL反应。测量火鸡单核细胞吞噬细胞氧化活性的可能性为研究火鸡单核细胞介导的宿主防御提供了一个有用的工具。

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