动物特异性CpG ODN的设计、筛选及对疫苗的增效作用

发布时间：2018-01-05 23:12

本文关键词：动物特异性CpG ODN的设计、筛选及对疫苗的增效作用　出处：《吉林大学》2010年博士论文　论文类型：学位论文

【摘要】： CpG ODN是含有非甲基化胞嘧啶—鸟嘌呤二核苷酸的单链脱氧寡核苷酸,通过与TLR9结合,能刺激人及哺乳动物的天然免疫系统,诱导产生以Th1型为主的免疫应答。CpG ODN作为人用疫苗佐剂在临床实验中取得了可喜的成果,作为动物疫苗佐剂的研究较少,这是因为CpG ODN具有种属特异性,即对一种动物表现生物活性的CpG ODN,在另一种动物则未必表现出同样的或同样强度的生物学活性。实验证明以CpG为核心的六核苷酸即CpG基序是导致种属特异性的原因。为了获得对牛或犬有免疫刺激作用的CpG ODN,以作为动物疫苗新型佐剂,我们根据CpG基序设计了60多条含有各种CpG基序的CpG ODN,经过体外筛选,获得了对犬免疫细胞有刺激活性的YW07、YW08和YW09；将YW07与含有铝佐剂的狂犬灭活疫苗联用免疫小鼠及犬,YW07能显著提高抗体产生水平及持续时间,并偏向Th1型免疫应答,且完全保护小鼠抵抗狂犬病毒的攻击。同时,获得了对牛免疫细胞有刺激活性的RW12,将RW12与含有206油佐剂的口蹄疫重组蛋白A7疫苗联用免疫小鼠及牛,RW12能显著提高其在动物体内的抗体水平及持续时间,一次免疫84天后进行攻毒实验,A7+206油+RW12组的牛80%(4／5)得到保护,灭活疫苗组只有50%(2／4)的牛得到保护。综上所述,我们开发的牛、犬特异性CpG ODN有望成为动物疫苗的新型高效佐剂。
[Abstract]:CpG ODN is a single stranded oligonucleotides containing unmethylated cytosine guanine dinucleotide -, combined with TLR9, can stimulate the innate immune system of human and other mammals, to induce Th1 type immune response.CpG ODN as vaccine adjuvant yielded promising results in clinical trials, as adjuvants of animal vaccine less this is because, CpG ODN has the species specificity, namely a kind of animal performance of the biological activity of CpG ODN, in another animal may not exhibit the same biological activity or the same strength. Results showed that six nucleotides with CpG as the core CpG motif is the cause of species specificity.
In order to obtain bovine or dog has the immunostimulatory effects of CpG ODN, as a new animal vaccine adjuvant, we designed more than 60 containing CpG motif of CpG ODN according to the CpG motif, after screening in vitro, obtained the stimulating activity of YW07 on canine immune cells, YW08 and YW09; YW07 and containing aluminum adjuvant the inactivated rabies vaccine combined with immune mice and dogs, YW07 could significantly increase the antibody level and duration, and bias of Th1 immune response, and completely protects mice against rabies virus attacks.
At the same time, the bovine immune cells stimulates the activity of RW12, RW12 and containing 206 oil adjuvant FMD recombinant protein A7 vaccine immunized mice and cattle, RW12 can significantly improve the antibody level of animal immunization and duration, 84 days after the poison attack experiment, 80% bovine A7+206 oil the +RW12 group (4 / 5) to be protected, the inactivated vaccine group was only 50% (2 / 4) of the cattle are protected.
In summary, the bovine, dog specific CpG ODN, which we have developed, is expected to be a new and highly efficient adjuvant for animal vaccines.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R392.9

【引证文献】