张柳,曹丽萍,丁炜东,熊传喜,殷国俊

• 1:华中农业大学水产学院
• 2:中国水产科学研究院淡水渔业研究中心
• 3:农业部水生动物遗传育种和养殖生物学重点开放实验室

摘要(Abstract)：

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。

关键词(KeyWords)： CpG-DNA;异育银鲫;巨噬细胞;外周血白细胞;免疫活性

Abstract:

Keywords:

基金项目(Foundation): 中国水产科学研究院淡水渔业研究中心中央级公益性科研院所基本科研业务费专项资金项目(No.6115009,6115038);; 江苏省国际科技合作计划(BZ2008008)

作者(Author): 张柳,曹丽萍,丁炜东,熊传喜,殷国俊

参考文献(References)：

• [1]Tokunaga T,Yamamoto H,Shimada,et al.Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG.I.Isolation,physicochemical characterization and antitumor activity[J].J Natl Cancer Inst,1984,72(4):955-962.
• [2]Cao H,Wang Y.Quantification of oxidative single-base and intrastrand cross-link lesions in unmenthylated and CpG-meethylated DNA induced by Fenton-type reagents[J].Nucleic Acids Res,2007,35(14):4833-4844.
• [3]Yamamoto S,Yamamoto T,Tokunaga T,et al.DNA from bacteria,but not from vertebtates,induces interferons,activitys natural killer cells and inhibits tumor growth[J].Microbiol Immunol,1992,36(9):983-997.
• [4]司兴奎,陈建红,张济培,等.CpG寡脱氧核苷酸的免疫刺激特性及应用研究进展[J].动物医学进展,2008,29(7):71-75.
• [5]Meng Z,Shao J Z,Xiang L X.CpG oligodeoxynucleotides activate grass carp(Ctenopharyngodon idellus)macrophages[J].Dev Comp Immunol,2003,27(4):313-321.
• [6]Tassakka A C M A R,Sakai M.Current research on the immunostimulatory effects of CpG oligodeoxynucleotides in fish[J].Aquaculture,2005,246(1-4):23-36.
• [7]Asmi Citra,Malina A R Tassakkaa,et al.Current research on the immunostimulatory effects of CpG oligodeoxynucleotides in fish[J].Aquaculture,2005(246):25-36.
• [8]丁航,侯敢,周克元.香菇多糖对巨噬细胞一氧化氮和一氧化氮合酶活性的影响[J].广东药学,2003,13(4):32-33.
• [9]Rosa D S,Bastos K R,Bargieri D Y,et al.Role of interferon-gamma during CpG oligodeoxynucleotide-adjuvanted immunization with recombinant proteins[J].Vaccine,2007,25(32):6007-6017.
• [10]田兴山,张玲华,邝哲师,等.不同剂量CpG序列对仔猪的免疫增强效果[J].华南农业大学学报(自然科学版),2004,25(1):96-98.
• [11]Dorn A,Keppenberger S.Clincal application of CpG-,non-CpCr-,and antisense oligodeoxynucleotides as immunomodulators[J].Curt Opin Mol Ther,2008,10(1):10-20.
• [13]Hartmann G,Krieg A M,Waldschmidt T J,et al.Mechanism and function of a newly identified CpG DNA motify in human primary B cells[J].J Immunol,2000,164:944-952.
• [12]Wu A,Oh S,Gharagozlou S,et al.In vivo vaccination with tumor cell lysate plus CpG oligodeoxynucleotides eradicates murine glioblastoma[J].J Immunother,2007,30(8):789-797.
• [14]Bayne C J.Pronephric leucocytes of Cyprinus carpio:isolation,separation and characterization Veterinary immunology and immu-nopathology,1986,12:141-151.
• [15]Yi A K,Klinman D M,Martin T L,et al.Rapid immune activation by CpG motifs in bacterial DNA.Systemic induction of IL-6transcription through an antioxidant-sensitive pathway[J].J Immunol,1996,157:5394-5402.