Desmidt M, Van Nerom A, Haesebrouck F, Ducatelle R, Ysebaert M T
Department of Pathology, Bacteriology and Poultry Diseases, Faculty of Veterinary Sciences, University of Gent, Belgium.
Vet Immunol Immunopathol. 1996 Oct;53(3-4):303-11. doi: 10.1016/S0165-2427(96)05620-6.
Luminol- and lucigenin-dependent chemiluminescence (CL) was used to compare activation of the respiratory burst of chicken peripheral blood monocytes and heterophils after stimulation with various agents. Monocytes and heterophils were obtained from the blood of three specific-pathogen-free chickens at 14 months of age and purified by a two-step discontinuous Percoll gradient. All cells responded to phorbol 12-myristate 13-acetate (PMA), zymosan A and calcium ionophore A23187 producing CL. The time course of luminol- and lucigenin-dependent CL was similar for both monocytes and heterophils after stimulation with PMA or zymosan A. Heterophils at lower cell number than monocytes responded with similar or higher peak maximum (PM) values. At the concentrations of stimuli used, the order of mean PM values was: zymosan A > PMA > A23187. Addition of 4 x 10(-6) M N-formyl-1-L-methionyl-L-leucyl-L-phenylalanine (fMLP) showed weak but significant CL activity at 1 x 10(6) monocytes per tube with luminol and at 5 x 10(6) monocytes per tube with lucigenin. No significant response to fMLP was observed with heterophils. The results indicate that the respiratory burst of chicken monocytes and heterophils can be measured by CL.
使用鲁米诺和光泽精依赖性化学发光(CL)来比较用各种试剂刺激后鸡外周血单核细胞和异嗜性粒细胞呼吸爆发的激活情况。单核细胞和异嗜性粒细胞取自3只14月龄无特定病原体鸡的血液,并通过两步不连续 Percoll 梯度进行纯化。所有细胞对佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)、酵母聚糖A和钙离子载体A23187均有反应,产生CL。在用PMA或酵母聚糖A刺激后,单核细胞和异嗜性粒细胞的鲁米诺和光泽精依赖性CL的时间进程相似。细胞数量低于单核细胞的异嗜性粒细胞产生的峰值最大值(PM)相似或更高。在所使用的刺激物浓度下,平均PM值的顺序为:酵母聚糖A > PMA > A23187。添加4×10⁻⁶ M N - 甲酰 - 1 - L - 甲硫氨酰 - L - 亮氨酰 - L - 苯丙氨酸(fMLP)时,每管1×10⁶个单核细胞在用鲁米诺时显示出微弱但显著的CL活性,每管5×10⁶个单核细胞在用光泽精时显示出微弱但显著的CL活性。异嗜性粒细胞对fMLP未观察到显著反应。结果表明,鸡单核细胞和异嗜性粒细胞的呼吸爆发可以通过CL来测量。