草鱼树突状细胞的分离鉴定及益生芽孢杆菌对其免疫功能的影响

为揭示草鱼(Ctenopharyngodon idellus)树突状细胞(Dendritic cells)的生物学特性及益生芽孢杆菌对其免疫功能的影响, 研究通过草鱼体外细胞培养技术分离获得草鱼DCs, 对其形态学特征、生物学功能及膜表面标记分子的表达进行了分析鉴定; RT-PCR检测了益生芽孢杆菌对草鱼DCs免疫相关细胞因子表达的影响。结果表明草鱼DCs具有典型的树突状形态, 可有效地激活T淋巴细胞的增殖并具有迁移能力。LPS刺激可促进其成熟过程, 显著提升膜表面标记分子CD80/86、CD83的表达。这表明草鱼DCs和哺乳动物DCs在形态和功能上具有高度相似性。RT-PCR结果显示, 在体外条件下使用紫外照射灭活的益生枯草芽孢杆菌对草鱼DCs进行刺激后, 抗炎性因子IL-4, IL-10的表达水平显著提升(P<0.05), 并在12h时达到峰值。这表明枯草芽孢杆菌可通过促进树突状细胞分泌抗炎性因子来影响其免疫功能。以上结果为进一步研究鱼类树突状细胞的生物学特性及益生芽孢杆菌对其免疫功能的影响提供了重要的依据。

IDENTIFICATION OF GRASS CARP DENDRITIC CELLS (CTENOPHARYNGODON IDELLUS) AND REGULATORY EFFECTS OF BACILLUS SUBTILIS ON THE IMMUNE FUNCTIONS

To investigate the biological characteristics of grass carp dendritic cells (Ctenopharyngodon idellus) and the regulatory mechanisms of dendritic cells by Bacillus subtilis, we isolated and gained grass carp dendritic cells to analyze morphological features, unique biological functions and authorized surface molecular markers. The expression of immune-related cytokines of dendritic cell to Bacillus subtilis stimulation was analyzed by RT-PCR. Our results showed that DCs have classical dendritic morphological features to initiate T cell proliferation and migration ability. LPS stimulation promoted the maturation progress of dendritic cells by increasing the expression of surface molecular markers CD83 and CD80/86, suggest that grass carp DCs were morphological and functional homology to mammalian dendritic cells. The expression of anti-inflammatory factors Il-4 and Il-10 increased significantly after the stimulation of UV-killed Bacillus subtilis (P<0.05), and reached to the highest level at 12h, revealing that Bacillus subtilis can promote the expression of anti-inflammatory factors to regulate the immune functions of dendritic cell. In conclusion, these results explored the biological characteristics of teleost fish dendritic cells and characterized the influence of probiotics Bacillus subtilis on the immune functions.