Studies on non-specific defence mechanism to bacterial infections in fishes.

研究了通过替代途径激活的鳗鱼补体的生物活性。将正常鳗鱼血清与酵母菌混合，酵母菌被认为是该途径的激活剂，或选定细菌的甲酸盐灭活细胞，并在25摄氏度下孵育60分钟。用离心法从酵母多糖或细菌细胞中分离出血清。首先，用改进的Boyden小室方法测定处理后血清的趋化活性。鳗鱼中性粒细胞向处理血清迁移的距离与向热灭活血清迁移的距离显著不同。其次，用处理后的血清给鳗鱼腹腔注射，以增强其诱导白细胞的活性。鳗鱼外周血中中性粒细胞数量迅速增加，在注射后6-9小时达到高峰。除了我们早期工作中展示的杀菌活性外，鳗鱼补体的这些趋化和白细胞诱导活性在TH…中也很重要更多针对细菌感染的非特异性防御。从鳗鱼外周血中分离出中性粒细胞。将1毫升未稀释的血液小心地装载到密度为1.073、1.083和1.089的不连续密度的菲科尔-偏三甲酸盐溶液中，并以1500rpm的速度将试管离心30min。在采血前一天，从注射了细菌死亡细胞的鳗鱼血液中获得了含有较重中性粒细胞的一部分。向鳗鱼注射处理过的SERE、酪蛋白或生理盐水(对照组)，并不能获得“重”的中性粒细胞。用化学发光法(CL)测定鳗鱼外周血中中性粒细胞的吞噬功能。中性粒细胞暴露于调理细菌后，CL反应迅速增加。对未调理细菌的CL反应明显低于调理细菌。CL反应量与正常血清作为调理素的浓度成正比。鲤鱼中性粒细胞吞噬的CL反应与鳗鱼相似，但强度低得多。结果表明，化学发光分析法不仅可以用来研究吞噬细胞的大小和动力学，还可以用来评价血清的调理活性。较少

Biological activities of eel complement activated through the alternative pathway were investigated. The normal eel sera were mixed with zymozan, which is known as an activator for the pathway, or formarin-killed cells of selected bacteria and incubated at 25 C for 60 min. The sera were separated from zymosan or the bacterial cells by centrifugation. First, chemotactic activity of the treated sera were determined by a modified Boyden chamber method. The distance of eel neutrophil migration toward the treated sera were significantly different from that toward the heat-inactivated sera. Secondly, leukocytosis-inducing activity was exzamined by intraperitoneally injecting eel with the treated sera. A rapid increase in the number of peripheral neutrophils was observed in the eel and reached a peak 6 - 9 hour after injection. In addition to bactericidal activity demonstrated in our earlier work these chemotactic and leukocytosis-inducing activities of eel complement appeared important in th … More e non-specific defence against bacterial infections.Neutrophils were seperated from the peripheral blood of eel. One ml of undiluted blood were carefully loaded onto a discontinuous density Ficoll-metrizoate ( 1.073, 1.083 and 1.089 in density) solution and the tube was centrifuged for 30 min at 1500 rpm. A fraction containing heavier neutrophils was obtained from the blood of eel injected with bacterial dead cells one day befor blood collection. The 'heavy' neutrophils were not obtained from eel injected with the treated sere, casein or saline (control). They were more active in phagacytosis than the ordinal 'light'ones.Phagocytosis of eel neutrophils isolated from peripheral blood was measured by chemiluminescence (CL) assay. When neutrophils were exposed opsonized bacteria, CL respones increased rapidly. CL response to the non-opsonized bacteria was much lower than that to the opsonized ones. The amount of CL respones became larger in proportion to the concentration of normal serum as an opsonin. CL response in phagocytosis of carp neutrophils was similar to that of eel in pattern, but much lower in intensity. It was found that the CL assay could be applied not only to investigate the magnitude and kinetics of phagaocyte but also to assess the serum opsonic activity. Less