CpG-DNA对异育银鲫(Carassius auratus gibelio)免疫细胞活性的诱导作用

探索含CpG基序的未甲基化寡脱氧核苷酸(ODNs)对鱼类的免疫调节,将人工合成的鱼免疫刺激剂寡脱氧核苷酸序列1670(含一个CpG基序)、1670-D(含两个CpG基序)加入到离体培养的异育银鲫的头肾巨噬细胞和外周血白细胞中,采用MTT法测定其对鲤外周血白细胞增殖的影响,NBT还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响。结果显示,1670可显著(浓度为20.0μg·mL-1)或极显著(浓度为0.1、1.0、10.0μg·mL-1)提高异育银鲫巨噬细胞的氧呼吸爆发活性,但对氮呼吸爆发活性和外周血白细胞的增殖无显著作用;1670-D可显著(浓度为0.1μg·mL-1)或极显著(浓度为1.0、10.0、20.0μg·mL-1)增强异育银鲫巨噬细胞的氧呼吸爆发活性和外周血白细胞的增殖,且可极显著(浓度为1.0、10.0、20.0μg·mL-1)提高异育银鲫巨噬细胞的氮呼吸爆发活性。表明特定序列、特定作用剂量的CpG-DNA可增强体外培养的异育银鲫巨噬细胞和外周血白细胞的非特异性免疫应答。

Effect of CpG-DNA on the Activation of Immunological Cells in the Carassius auratus gibelio

To investigate the immunomodulating effects of oligodeoxynucleotides (ODNs) containing unmethylated CpG in fish,two synthetic ODNs,1670 containing one CpG basal sequence and 1670-D containing two CpG basal sequences,were added to the macrophages and the peripheral blood leukocytes isolated from the head kidney and blood of Carassius auratus gibelio respectively. The proliferation of the peripheral blood granulocytes,respiratory burst of macrophage were analyzed by MTT,NBT reduction,Griess reaction. Results showed that 1670 at 20 μg · mL-1 (P < 0.05) and 0.1,1.0,10 μg · mL-1(P < 0.01) could significantly stimulate oxygen burst activity of macrophages,but it had no effects on the nitrogen burst activity of macrophage and the proliferation of the peripheral blood granulocytes. 1670-D at 0.1 μg · mL-1 (P < 0.05) and 1.0,10.0,20.0 μg · mL-1 (P < 0.01) could significantly stimulate oxygen burst activity of macrophages and the proliferation of the peripheral blood granulocytes; at 1.0,10.0 and 20.0 μg · mL-1,it could also very significantly stimulate the nitrogen burst activity of macrophage. The results showed that,at the suitable dose,the specific ODNs could stimulate the non-specific immune responses of the macrophages and the peripheral blood leukocytes in fish. The indifferential immunity of the head kidney macrophages and peripheral blood granulocytes could be advanced with the specifical doses of the specifical sequences.