Prototype development and preclinical immunogenicity analysis of a novel minimally invasive electroporation device
Prototype development and preclinical immunogenicity analysis of a novel minimally invasive electroporation device
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DOI：
10.1038/gt.2010.137
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发表时间：
2011-03-01
期刊：
影响因子：
5.1
通讯作者：
Sardesai, N. Y.
中科院分区：
文献类型：
--
作者：
Broderick, K. E.;Shen, X.;Sardesai, N. Y.
对DNA疫苗的免疫应答的大小取决于三个标准-优化的载体设计、合适佐剂的使用以及质粒在靶组织中的成功递送和随后表达。体内电穿孔（EP）已被证明在将DNA免疫原有效递送至肌肉和皮肤方面特别有效，并且实际上几种装置已进入人体临床试验。在这里，我们报告了一个新的概念，DNA输送到皮肤组织使用微创EP设备，这是使用低电压参数供电。我们表明，这个原型装置含有一个新的4x 4电极阵列的结果在鲁棒性和可重复的转染真皮组织和随后的抗原表达在注射部位。使用编码NP和M2 e流感抗原的DNA，我们进一步显示了在小鼠模型中诱导有效的细胞应答，如通过抗原特异性T细胞ELISpot测定所测量的。重要的是，当用VN/1203/04（H5 N1）流感毒株攻击时，100%的免疫动物受到保护。我们还将我们的发现扩展到豚鼠模型，并证明了针对大流行性新型H1N1病毒的HI滴度大于1：40的诱导，显示了在广谱物种中使用原型装置并使用多种抗原进行DNA递送的概念效力的证明。最后，我们能够在猕猴中产生针对相同的新型H1N1毒株的保护性HI滴度。我们的研究结果表明，微创皮肤装置可以提供一种安全，耐受和有效的方法来管理DNA疫苗的预防性设置，因此可能代表了一个重要的新的选择，改善DNA疫苗在体内的交付。Gene Therapy（2011）18，258-265; doi：10.1038/gt.2010.137; 2010年10月21日在线发表
The magnitude of the immune response to a DNA vaccine depends on three criteria-the optimized vector design, the use of a suitable adjuvant and the successful delivery and subsequent expression of the plasmid in the target tissue. In vivo electroporation (EP) has proved to be particularly effective in efficiently delivering DNA immunogens to the muscle and the skin, and indeed several devices have entered into human clinical trials. Here, we report on a novel concept of DNA delivery to the dermal tissue using a minimally invasive EP device, which is powered using low-voltage parameters. We show that this prototype device containing a novel 4x4-electrode array results in robust and reproducible transfection of dermal tissue and subsequent antigen expression at the injection site. Using DNA encoding for NP and M2e influenza antigens, we further show induction of potent cellular responses in a mouse model as measured by antigen-specific T-cell ELISpot assays. Importantly, 100% of the immunized animals were protected when challenged with VN/1203/04 (H5N1) strain of influenza. We have also extended our findings to a guinea-pig model and demonstrated induction of HI titers greater than 1: 40 against a pandemic novel H1N1 virus showing proof of concept efficacy for DNA delivery with the prototype device in a broad spectrum of species and using multiple antigens. Finally, we were able to generate protective HI titers in macaques against the same novel H1N1 strain. Our results suggest that the minimally invasive dermal device may offer a safe, tolerable and efficient method to administer DNA vaccinations in a prophylactic setting, and thus potentially represents an important new option for improved DNA vaccine delivery in vivo. Gene Therapy (2011) 18, 258-265; doi:10.1038/gt.2010.137; published online 21 October 2010