不同序列的CpG-ODN对猪体外免疫细胞免疫刺激作用的研究

为了筛选出对猪体有免疫刺激活性的CpG寡聚脱氧核苷酸（CpG-ODN），人工设计合成了9种未甲基化CpCr-ODN不同序列，分别作用于健康猪外周血淋巴细胞和脾脏免疫细胞，进行体外转化试验。用四甲基偶氮唑盐（MTT）比色法检测外周血淋巴细胞转化效果和脾细胞的增殖能力和代谢能力，通过测得的OD490值，计算出淋巴细胞增殖指数（SI），筛选CpG-ODN的免疫刺激作用。结果显示：9种不同序列的CpG-ODN对猪外周血淋巴细胞和脾脏免疫细胞均具有不同程度的刺激作用，其中CpG-ODN7对淋巴细胞增殖能力的增强效果最为明显（sI=3．5），CpG-ODN6对猪脾脏免疫细胞的刺激指数最大（SI=2．7）。为研制猪DNA疫苗有效的CpG-ODN免疫佐剂奠定了基础。

The Preliminary Study on Different CpG-ODN Sequences with Immunostimulatory in Vitro Immue Cells of Swine

In the study,nine different artificial unmethylated oligodexynucleotides(CpG-ODN) sequences to the swine peripheral blood lymphocytes and spleen cells respectively were made by artificial methods.The effect of the invitro lymphocyte transformation was detected by MTT assay.According to the tested OD,The stimulation index of CpG-ODN(SI) was screened out to study the immune effect of CpG-ODN.It is showed that the nine CpG-ODNs had different extent of immunostimulatory effects on the swine peripheral blood lymphocytes and spleen cells.After elaborate analysis,the conclusion was drawn that the CpG-ODN7 had the strongest immunostimulatory activity on the swine peripheral blood lymphocytes(SI=3.5) and CpG-ODN6 had the strongest immunostimulatory activity on the swine spleen cells(SI=2.7).This would be useful for study of effective CpG-ODN immune adjunct for gene vaccination in future.