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体外筛选对鸡具有免疫刺激活性的CpG寡脱氧核苷酸

DOI: 10.7685/j.issn.1000-2030.2004.02.020, PP. 83-86

Keywords: ,CpG寡脱氧核苷酸,CpG模体,淋巴细胞转化,免疫刺激活性

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Abstract:

分别用有丝分裂原和CpG寡脱氧核苷酸(CpGODN)对SPF鸡全血,经低速离心分离的外周血单个核细胞(PBMC)和脾脏细胞,用淋巴细胞分离液分离的PBMC和脾脏细胞进行刺激,用3HTdR掺入法测定并比较上述鸡免疫细胞在不同培养温度和细胞浓度下的转化效果,从而建立了适用于大批量筛选对鸡具有免疫刺激活性CpGODN的淋巴细胞转化方法。进一步用该方法筛选自行设计合成的38条ODN。结果表明,用低速离心法分离的鸡PBMC,在细胞含量为107mL-1、37℃、5%CO2条件下,经CpGODN刺激64~66h后,可以得到稳定的高效转化;不含CpG模体的ODN不具有免疫刺激活性;CpGODN10、11和14具有最佳的免疫刺激活性,刺激指数大于10。

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