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Skin-specific promoters for genetic immunisation by DNA electroporation

机译：通过DNA电穿孔进行皮肤免疫的皮肤特异性启动子

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摘要

This study aimed at restricting expression of DNA vaccine to specific cell types by using skin-specific promoters (i) to contribute to the understanding of the mechanism of intradermal DNA vaccines and (ii) to address safety concerns associated with DNA vaccines. The expression and immune response after delivery of plasmids encoding luciferase by intradermal DNA electroporation were assessed. Two ubiquitous promoters, CMV and CAG, and three tissue-specific promoters were studied. Keratin 14 promoter restricts gene expression to keratinocytes of the epidermal basal layer, CD11c promoter to dendritic cells and fascin promoter only to mature dendritic cells. The use of plasmids with tissue-specific promoter resulted in significant, but very low protein expression, as compared to that obtained with ubiquitous promoter plasmids. Immunisation with both ubiquitous promoter plasmids elicited humoral and cellular anti-luciferase immune response. No immune response was observed after delivery of CD11c plasmid while fascin and keratin 14 plasmids induced IFN-gamma response suggesting that the targeting of skin-specific cells could be a suitable approach but only for the treatment of pathologies or pathogens requiring mainly cellular and not humoral immune response.
机译：这项研究旨在通过使用皮肤特异性启动子将DNA疫苗的表达限制为特定细胞类型（i）有助于理解皮内DNA疫苗的机制，以及（ii）解决与DNA疫苗相关的安全性问题。评估通过皮内DNA电穿孔递送编码荧光素酶的质粒后的表达和免疫应答。研究了两个普遍存在的启动子，CMV和CAG，以及三个组织特异性启动子。角蛋白14启动子将基因表达限制在表皮基底层的角质形成细胞上，CD11c启动子限制在树突状细胞上，而fascin启动子仅限制在成熟的树突状细胞上。与具有普遍存在的启动子质粒的表达相比，具有组织特异性启动子的质粒的使用导致显着但非常低的蛋白质表达。两种普遍存在的启动子质粒的免疫均引起体液和细胞抗荧光素酶的免疫反应。递送CD11c质粒后未观察到免疫应答，而fascin和角蛋白14质粒诱导IFN-γ应答，提示靶向皮肤特异性细胞可能是合适的方法，但仅用于治疗主要需要细胞而非体液的病理或病原体免疫反应。

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