CpG寡脱氧核苷酸免疫刺激活性的种属特异性  

Heterogeneity in the Response of Different Animals to Specific CpG Oligodeoxynucleotides

作　　者：李光富[1] 朱鸿飞[2] 马艳琴[1] 朱建中[1] 邵国清[3] 卢春[1] 张兆松[1] 

机构地区：[1]南京医科大学医学分子生物学研究所,南京210029 [2]南京农业大学生物工程研究所,南京210095 [3]江苏省农业科学院兽医研究所,南京210095

出　　处：《农业生物技术学报》2004年第1期66-70,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展(863)计划(No.2001AA243071);国家自然科学资金项目(No.30271166)

摘　　要：研究了CpG寡脱氧核苷酸(CpGODN)对猪和鼠免疫刺激活性的差异。根据CpGODN的研究结果,筛选并化学合成了对人、猪和鼠分别有最佳免疫刺激活性的CpGODN2006、D19、1826和不含CpG基序的阴性对照ODND48。用3H-TdR掺入法测定CpGODN刺激淋巴细胞增殖的效果;用双抗体夹心ELISA测定刺激细胞上清的IL-2和IFN-γ水平;用SPSS软件对数值进行统计分析。结果表明,D19能强烈地诱发猪外周血单核细胞(PBMC)的增殖,且D19刺激猪PBMC细胞上清中IL-Ⅱ和IFN-γ水平明显升高,但不能有效地刺激鼠脾细胞的增殖;1826能强烈地诱发鼠脾淋巴细胞的增殖,但不能有效地刺激猪PBMC的增殖;2006能同样有效地诱发猪PBMC和鼠的脾淋巴细胞增殖,刺激猪PBMC增加IL-2的分泌。所以,CpGODN免疫刺激活性既具有一定的种属特异性,也存在一定的广谱性,D19更趋向于诱导机体T辅助细胞1(Th1)型免疫应答。The differences of CpG oligodeoxynucleotides(ODN) immunostimulation activity between different animal species were studied. Three kinds of CpG ODN 2 006, D19 and 1826 identified optimally activity in human, swine and mice were se lected and synthesized respectively, and so was the control ODN D48 which was no CpG-containing oligodeoxynucleotides. Swine peripheral blood mononuclear cells (PBMC) were prepared by density gradient centrifugation, mouse spleen lymphocy tes by resolving erythrocyte with NH4Cl; For in vitro proliferation assays, in corporation of 3H-thymidine was determined using standard protocols and stimulat ion index was calculated; and IFN-?and IL-2 production in activated cells were measured with swIFN-? and IL-2 kit which were a solid phase sandwich enzyme lin ked-immuno-sorbent assay(ELISA). Results indicated that D19 could strongly stimu late pig PBMC proliferation, and enhanced IFN-?production; but the mice sple en lymphocyte did not respond to it. In reverse, mice spleen lymphocytes stimnul ated with 1826 showed the obvious proliferation, but pig PBMCs did not respond t o it. But 2006 could effectively stimulate proliferation of both pigs PBMCs and mice spleen lymphocytes. So, some specific CpG ODNs could selectively stimulate animal species, and some possessed common stimulation effect; And D19 could ind uce predominantly the response of Th1 in vivo.

关 键 词：免疫 CPG基序 CPG寡脱氧核苷酸 免疫刺激活性 猪 鼠 

分 类 号：S852.4[农业科学—基础兽医学]