不同CpG-DNA序列对异育银鲫(Carassius auratus gibelio)离体培养免疫细胞活性影响的研究  

Investigation of the Effect of CpG-DNA on the Activity of Immunological Cells in vitro of Carassius auratus gibelio

作　　者：曹丽萍[1] 丁炜东[1] 张柳[1,2] 殷国俊[1] 

机构地区：[1]中国水产科学研究院淡水渔业研究中心,农业部水生动物遗传育种和养殖生物学重点开放实验室,江苏无锡214081 [2]华中农业大学水产学院,湖北武汉430070

出　　处：《云南农业大学学报（自然科学版）》2010年第5期629-635,共7页Journal of Yunnan Agricultural University:Natural Science

基　　金：中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院淡水渔业研究中心)项目(2007JBFA06,2007JBFC05);科技部国际科技合作项目(2009DFA32620);江苏省国际科技合作计划项目(BZ2008008);无锡市科技计划(国际科技合作)项目(CZE00906)

摘　　要：采用Percoll密度离心等技术,对异育银鲫的头肾巨噬细胞和外周血白细胞进行离体培养,经人工合成的CpG-DNA序列作用后,测定其对头肾巨噬细胞的氧、氮呼吸爆发活性和对外周血白细胞增殖的影响,从CpG-DNA核心CG两端的侧翼序列(种属)、核心CG的数量和位置等方面观察CpG-DNA基序对异育银鲫免疫细胞活性的影响。结果表明:1670较1826和2006具有更显著的增强巨噬细胞的氧呼吸爆发活性和白细胞的增殖作用(P<0.05);1670-N与同浓度1670比较具有更强的提高白细胞的增殖能力;1670-D比同浓度的1670更能显著地促进白细胞增殖和巨噬细胞的氮呼吸爆发活性(P<0.05);1670-R对巨噬细胞氧、氮爆发活性及外周血白细胞的增殖无显著促进作用。结果证明了CpG-DNA可以通过促进巨噬细胞的氮氧爆发活性和增强外周血白细胞的增殖来提高机体的非特异性免疫功能,同时这种作用受到核心CG两端的侧翼序列、核心序列的位置数量及核心CG顺序的影响。The head kidney macrophages and peripheral blood granulocytes isolated from Carassius auratus gibelio were purified by means of Percoll gradient density centrifugation and were exposed to artificial sequences of CpG-DNA in vitro.To evaluate the immunostimulating effects of CpG-DNAs and compare their immune activities with the control group,proliferation of the peripheral blood leukocytes,and burst of macrophage were investigated.Results showed that superoxide anion in macrophages and proliferation of peripheral blood leukocytes by the induction of 1670 were more markedly stimulated than 1826 and 2006（P0.05）.1670-N could induce superoxide anion in macrophages markedly much more,compared with the same concentrations of 1670.The detections of respiratory burst of macrophage and the peripheral blood leukocytes after incubation by 1670-D showed it had more conspicuous effect on immunocompetence than the same concentrations of 1670（P0.05）.But the effects were not obvious by 1670-R.It is suggested that the location and amount of the CpG-DNA had important effect on its immunocompetence.CpG-DNA could induce superoxide anion and had conspicuous effects on nitrogen the effects were influenced by the CG sequences,the two zygomorphic sequences of CG,the positions and the amounts of the core sequence.

关 键 词：CPG-DNA 巨噬细胞 氧爆发活性 氮爆发活性 外周血白细胞增殖 

分 类 号：S942.5[农业科学—水产养殖]