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设计并筛选对猪具有最佳免疫刺激活性的CpGODN。文献报道 ,设计并合成了 38条CpGODN。用猪PBMC体外增殖实验 ,3 H 胸腺嘧啶 (3 H TdR )掺入法测定刺激指数 ,用SPSS软件对数值进行统计学分析 ;用双抗体夹心ELISA检测PBMC培养上清中的IFN γ和IL 2的量 ,来评价CpGODN对猪的免疫学作用。结果表明 ,多数CpGODN能够刺激猪PBMC的增殖 ,并促进其分泌IFN γ ,但IL 2分泌的量均未显著增加。其中 ,含有三个GTCGTT基序ODN 2 0 0 6刺激猪PBMC增殖的作用最强 ,刺激指数达 6 2 ,但仅能诱生低水平的IFN γ ;具有磷酸二酯键 /硫代磷酸二酯键混合骨架及 3’端polyG尾的ODND1 9具有中等程度的促猪PBMC增殖的作用 ,刺激指数为 4 2 ,但诱导PBMC分泌IFN γ比阴性对照增加了 5倍。ODND1 9和ODN 2 0 0 6对猪具有不同的免疫学效应 ,这为进一步研究适用于不同病原体的猪用疫苗佐剂提供了实验参考
CpGODN was designed and screened for the best immunostimulatory activity in pigs. Reported in the literature, 38 CpGODNs were designed and synthesized. Stimulation index was determined by porcine PBMC in vitro proliferation assay with 3 H TdR incorporation method and SPSS software was used for statistical analysis. The IFNγ and IL-2 in supernatant of PBMC were detected by double antibody sandwich ELISA To evaluate the immunological effects of CpGODN on pigs. The results showed that most CpG ODN could stimulate the proliferation of pig PBMC and promote the secretion of IFN γ, but the amount of IL 2 secretion did not significantly increase. Among them, three GTCGTT motif ODN 2 0 0 6 stimulated the proliferation of porcine PBMC strongest, stimulating index of 6 2, but only induced low levels of IFN γ; with phosphodiester / phosphorothioate diester The ODND1 9 of the key hybrid backbone and the 3 ’polyG tail had a moderate effect on the proliferation of porcine PBMCs with a stimulation index of 42, but the induction of IFNγ by PBMCs was 5-fold more than the negative control. ODND1 9 and ODN 2 0 0 6 have different immunological effects on pigs, which provides experimental reference for further study on porcine vaccine adjuvants suitable for different pathogens