免疫调节性寡聚脱氧核苷酸筛选方法的建立、优化和应用

目的:建立一套合理而便捷的实验体系,为开发新型免疫调节性寡聚脱氧核苷酸(ODN)提供筛选方法。方法:利用含有CpG基序的免疫刺激性寡聚脱氧核苷酸(CpG ODN)作为刺激物,将人外周血单个核细胞(PBMC)作为研究对象,分别以细胞的增殖程度和刺激上清的抗病毒活性作为检测指标,优化各项实验条件,综合评定寡聚脱氧核苷酸的免疫调节活性。结果:成功建立了由阳性免疫调节性ODNA151抑制CpG ODN诱导的人PBMC增殖及抗病毒活性的筛选方法。结论:免疫调节性ODN筛选方法的成功建立,为下一步研究开发新型免疫调节性ODN奠定了基础。

Establishment,optimization and application of a screening method for immunoregulatory oligodeoxynucleotides

AIM:To establish a set of reasonable and convenient experimental system to provide a screening method for the development of novel immunoregulatory oligodeoxynucleotides.METHODS:The human PBMCs were stimulated by CpG ODN and/or immunoregulatory ODN.The cell proliferation and anti-viral activity of the supernatant induced by CpG ODN were examined by thymidine incorporation and anti-viral bioassay to evaluate the immunoregulatory activity of candidate ODN.The experimental conditions were also optimized.RESULTS:A screening method on which A151,a positive immunoregulatory ODN,inhibited the proliferation and anti-viral activity of the CpG ODN-induced human PBMCs was successfully established.CONCLUSION:The successful establishment of CpG ODN based screening method lays the foundations for further development of novel immunoregulatory oligodeoxynucleotides.