Jørgensen J B, Robertsen B
Department of Marine Biochemistry, Norwegian College of Fishery Science, University of Tromsø.
Dev Comp Immunol. 1995 Jan-Feb;19(1):43-57. doi: 10.1016/0145-305x(94)00045-h.
Previous studies have shown that head kidney macrophages isolated from glucan injected rainbow trout (Oncorhychus mykiss) and Atlantic salmon (Salmo salar L.) have increased ability to kill Aeromonas salmonicida. The present work was aimed at investigating the in vitro effects of glucan on the respiratory burst and bactericidal potential of Atlantic salmon head kidney macrophages. Salmon macrophages were incubated for 1-7 days with various concentrations of yeast beta-glucan (MacroGard) and tested for respiratory burst activity by the reduction of nitroblue-tetrazolium (NBT) after exposure to phorbol myristate acetate (PMA) or opsonized zymosan. The macrophages showed a marked increase in respiratory burst activity 4 to 7 days after addition of glucan. Macrophages treated with 0.1-1 microgram mL-1 gave a maximum respiratory burst response, whereas 10 micrograms mL-1 had no effect and 50 micrograms mL-1 was inhibitory. The glucan also triggered respiratory burst activity directly, but this occurred only at relative high concentrations with a maximal effect at > or = 200 micrograms mL-1. The validity of using the NBT-assay as a measure of respiratory burst activity was confirmed by using inhibitors of O2- production (superoxide dismutase, trifluoperazine and diphenylene iodonium). Despite the stimulatory effect of glucan on the respiratory burst activity of salmon macrophages, these cells did not show increased bactericidal activity against the avirulent and virulent strain of A. salmonicida. Upregulation of burst activity alone is thus apparently not sufficient to enhance bactericidal activity against this pathogen by Atlantic salmon macrophages.
先前的研究表明,从注射葡聚糖的虹鳟鱼(Oncorhychus mykiss)和大西洋鲑鱼(Salmo salar L.)中分离出的头肾巨噬细胞杀灭杀鲑气单胞菌(Aeromonas salmonicida)的能力增强。本研究旨在调查葡聚糖对大西洋鲑鱼头肾巨噬细胞呼吸爆发和杀菌潜力的体外影响。将鲑鱼巨噬细胞与不同浓度的酵母β-葡聚糖(MacroGard)孵育1 - 7天,并在暴露于佛波酯(PMA)或调理酵母聚糖后,通过硝基蓝四氮唑(NBT)还原试验检测呼吸爆发活性。添加葡聚糖后4至7天,巨噬细胞的呼吸爆发活性显著增加。用0.1 - 1微克/毫升处理的巨噬细胞产生最大的呼吸爆发反应,而10微克/毫升无作用,50微克/毫升具有抑制作用。葡聚糖也直接引发呼吸爆发活性,但这仅在相对高浓度下发生,在≥200微克/毫升时具有最大效应。通过使用O2-产生抑制剂(超氧化物歧化酶、三氟拉嗪和二苯碘鎓)证实了使用NBT测定法作为呼吸爆发活性测量方法的有效性。尽管葡聚糖对鲑鱼巨噬细胞的呼吸爆发活性有刺激作用,但这些细胞对杀鲑气单胞菌的无毒和有毒菌株均未表现出增强的杀菌活性。因此,仅呼吸爆发活性的上调显然不足以增强大西洋鲑鱼巨噬细胞对该病原体的杀菌活性。