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The production and immunostimulatory activity of double-stranded CpG-DNA

机译:双链CPG-DNA的生产和免疫刺激活性

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CpG-DNA, which contains unmethylated CpG dinucleotides in the context of specific sequences, has remarkable and diverse immunological effects, including induction of proinflammatory cytokine expression and regulation of the Th1/Th2 immune response. Here, we examined the immunostimulatory activities of double-stranded (ds) CpG-DNA in the human B cell line RPMI8226. To investigate whether dsCpG-DNA stimulates immune cells, we constructed a plasmid containing repeated dsCpG-DNA and produced dsCpG-DNA by PCR amplification and EcoR I digestion. PCR-amplified dsCpG-DNA alone did not have immmunostimulatory activity. However, dsCpGDNA encapsulated with lipofectin induced IL-8 promoter activation, HLA-DRA expression, and IL-8 expression in a CG sequence- independent manner. The effects of encapsulated dsCpGDNA were independent of minor endotoxin contamination. These findings suggest the potential use of dsCpG-DNA as a therapy for immune response regulation.
机译:在特定序列的背景下含有未甲基化的CPG二核苷酸的CpG-DNA具有显着且不同的免疫效应,包括诱导促炎细胞因子表达和调节TH / TH2免疫应答。在这里,我们检查了人B细胞系RPMI8226中双链(DS)CpG-DNA的免疫刺激活性。为了研究DSCPG-DNA是否刺激免疫细胞,我们构建了一种含有重复的DSCPG-DNA的质粒,并通过PCR扩增和ECOL I消化产生DSCPG-DNA。单独的PCR扩增的DSCPG-DNA没有免疫刺激性活性。然而,以CG序列独立的方式封装脂素蛋白诱导的IL-8启动子活化,HLA-DRA表达和IL-8表达。包封的DSCPGDNA的效果与次要内毒素污染无关。这些发现表明DSCPG-DNA的潜在用途作为免疫应答调节的疗法。

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