Huttenhuis Heidi B T, Ribeiro Ana S P, Bowden Timothy J, Van Bavel Casandra, Taverne-Thiele Anja J, Rombout Jan H W M
Cell Biology and Immunology Group, Wageningen Institute of Animal Sciences, Wageningen University, Marijkeweg 40, 6709 PG Wageningen, The Netherlands.
Fish Shellfish Immunol. 2006 Sep;21(3):261-71. doi: 10.1016/j.fsi.2005.12.002. Epub 2006 Feb 14.
The effect of a 2-week period of oral immuno-stimulation from the age of 2 or 6 weeks post-fertilisation (wpf; before and after reaching the ability to produce antibodies) onwards was investigated on various immune functions of the common carp, Cyprinus carpio. The immuno-stimulants Aeromonas salmonicida lipopolysaccharide, Yeast DNA (containing unmethylated CpG motifs) or high-M alginate (an extract of algae containing poly-mannuronic acid) were used. The effect of this treatment was studied on the kinetics of B cells in head kidney and peripheral blood leucocytes using flow cytometry, on the total plasma IgM level using ELISA, on cytokine and inducible nitric oxide synthase (iNOS) expression in the intestine, and acute phase protein expression in the liver, using real time quantitative PCR, and on exposure to Vibrio anguillarum. Oral administration of immuno-stimulants from 6 wpf resulted in decreased WCI12(+) (B) cell percentages in PBL (only after administration of LPS) and head kidney (all test groups), and a decreased total IgM level in plasma, suggesting that suppressive effects are strongly indicative of oral or juvenile tolerance. After administration from 2 wpf, the effects on WCI12(+) (B) cell percentages were less pronounced: the group fed with Yeast DNA showed higher percentages compared to the control group at 6 wpf, but lower percentages at 8 wpf. No changes were observed in the cytokine or iNOS expression levels in the intestine or acute phase protein expression in the liver. A challenge with V. anguillarum resulted in an initially higher cumulative mortality in the group fed with LPS, but lower mortality in the groups fed with Yeast DNA or high-M alginate compared to the control group, providing a provisional warning especially for the use of pathogen-derived immuno-stimulants, such as A. salmonicida LPS, in larval and juvenile fish.
研究了从受精后2周或6周龄（wpf；在达到产生抗体能力之前和之后）开始进行为期2周的口服免疫刺激对鲤鱼（Cyprinus carpio）各种免疫功能的影响。使用了免疫刺激剂杀鲑气单胞菌脂多糖、酵母DNA（含有未甲基化的CpG基序）或高M海藻酸盐（一种含有聚甘露糖醛酸的藻类提取物）。使用流式细胞术研究了这种处理对头肾和外周血白细胞中B细胞动力学的影响，使用酶联免疫吸附测定法研究了血浆总IgM水平，使用实时定量聚合酶链反应研究了肠道中细胞因子和诱导型一氧化氮合酶（iNOS）的表达以及肝脏中急性期蛋白的表达，并研究了对鳗弧菌感染的反应。从6 wpf开始口服免疫刺激剂导致外周血淋巴细胞（PBL，仅在给予脂多糖后）和头肾（所有试验组）中WCI12(+)（B）细胞百分比降低，以及血浆中总IgM水平降低，这表明抑制作用强烈表明存在口服或幼年耐受性。从2 wpf开始给药后，对WCI12(+)（B）细胞百分比的影响不太明显：喂食酵母DNA的组在6 wpf时比对照组百分比更高，但在8 wpf时更低。在肠道中细胞因子或iNOS表达水平以及肝脏中急性期蛋白表达方面未观察到变化。用鳗弧菌进行攻击导致喂食脂多糖的组最初累积死亡率较高，但与对照组相比，喂食酵母DNA或高M海藻酸盐的组死亡率较低，这尤其为在幼鱼中使用病原体衍生的免疫刺激剂（如杀鲑气单胞菌脂多糖）提供了一个临时警示。
