Nikoskelainen Sami, Lehtinen Janne, Lilius Esa-Matti
Department of Biochemistry and Food Chemistry, University of Turku, 20014 Turku, Finland.
Dev Comp Immunol. 2002 Nov;26(9):797-804. doi: 10.1016/s0145-305x(02)00032-0.
The total bacteriolytic activity comprising of the classical, alternative and possible lectine pathways as well as the bacteriolytic activity of the alternative pathway (AP) of rainbow trout (Oncorhynchus mykiss) complement was assessed in temperatures ranging from 0 to 35 degrees C against a recombinant strain Escherichia coli containing two reporter genes gfp and lucFF. At 35 degrees C there was no difference between the total (TC) activity and the activity of the AP, but at 10 degrees C the TC was notably higher than the AP. Total activity peaked at 30 degrees C and gradually grew smaller towards 0 degrees C. The activity of the AP was similarly temperature-dependent, but CB50 value was found to be beyond measurable range at temperatures below 10 degrees C. When compared to human serum complement, the peak human TC activity at 37 degrees C was four times higher than the TC of rainbow trout at 30 degrees C. Human TC activity was 10.1-fold lower at 25 degrees C when compared to the activity at 37 degrees C. At 37 degrees C the human AP bacteriolytic activity was 4.5-fold less effective than human TC, but at 25 degrees C there was no difference between human TC and AP. In contrast to previous reports where AP activity of fish was assayed as hemolytic activity our study showed that the bacteriolytic activity of AP was lower than that of TC and very low at temperatures below 10 degrees C suggesting that the earlier proposed particular importance of AP in fish should be reconsidered.
在0至35摄氏度的温度范围内，针对含有两个报告基因gfp和lucFF的重组大肠杆菌菌株，评估了虹鳟（Oncorhynchus mykiss）补体经典途径、替代途径和可能的凝集素途径的总溶菌活性以及替代途径（AP）的溶菌活性。在35摄氏度时，总（TC）活性与AP活性之间没有差异，但在10摄氏度时，TC明显高于AP。总活性在30摄氏度时达到峰值，并朝着0摄氏度逐渐降低。AP的活性同样依赖于温度，但在低于10摄氏度的温度下，CB50值超出了可测量范围。与人类血清补体相比，人类TC在37摄氏度时的峰值活性比虹鳟在30摄氏度时的TC高四倍。与37摄氏度时的活性相比，人类TC在25摄氏度时的活性低10.1倍。在37摄氏度时，人类AP的溶菌活性比人类TC低4.5倍，但在25摄氏度时，人类TC和AP之间没有差异。与之前将鱼类AP活性测定为溶血活性的报道不同，我们的研究表明，AP的溶菌活性低于TC，并且在低于10摄氏度的温度下非常低，这表明之前提出的AP在鱼类中特别重要的观点应该重新考虑。
