Cutaneous DNA immunization with polymer-nanoparticles

描述(由申请人提供)：基于DNA的疫苗有可能对各种疾病非常有效。它们也是目前可用的疫苗的更便宜、更容易大规模生产和潜在更安全的替代品。尽管最近在各种动物模型中取得了成功，但基因免疫在人类身上的预防和治疗成功还远远没有实现，还有待于开发出符合患者要求的、高效、安全和实用的基因递送系统。通过皮肤的无针免疫是目前给药方案的一种极好的替代方案，因为：(A)皮肤是抗原提呈细胞的丰富环境，这些细胞天生就是为免疫监测和调节而设计的，(B)它是针刺注射的更安全和更符合患者要求的替代方案。使用高压介导的生物制剂或粉末递送的皮肤免疫的最新进展，已被证明在几种疾病模型中是有效的。然而，这些疫苗需要高度专业的设备和熟练的人员，这就排除了它们用于大规模疫苗接种的可能性，特别是在第三世界国家。使用DNA纳米颗粒的自我适用、基于贴片的递送技术可以显著降低成本，但却是高效的皮肤疫苗接种策略。这项提案描述了收集关于通过聚合物纳米颗粒和化学增强剂传递的质粒DNA和CpG寡核苷酸对皮肤渗透和随后产生针对编码抗原的免疫反应的有效性的初步数据。其具体目的是：(A)合成不同尺寸和不同表面电荷的可生物降解纳米颗粒，将质粒DNA和CpG寡聚糖吸附在表面并与不同的透皮剂复合，(B)研究化学增强剂和离子导入给药后的颗粒转运和树突状细胞摄取的有效性，以及(C)贴片皮肤应用后对编码抗原的免疫应答。这些研究将使我们能够确定皮肤颗粒递送的关键参数，并进一步优化和开发一种简单、无针、皮肤贴片类型的系统，用于特定疾病模型的DNA免疫。

DESCRIPTION (provided by applicant): DNA-based vaccines have the potential to be highly effective in various diseases. They are also less expensive, easy to mass produce and potentially safer alternative to currently available vaccines. Despite the recent success in various animal models, the prophylactic and therapeutic success of genetic immunization in humans is far from realization and awaits development of patient-compliant, efficient, safe, and practical gene delivery systems. Needle-free immunization through the skin is an excellent alternative to current delivery protocols because, (a) the skin is a rich environment for antigen presenting cells that are naturally designed for immune-surveillance and modulation and (b) it is a safer and more patient-compliant alternative to needled injections. Recent developments in cutaneous immunization using high-pressure mediated biolistic or powder delivery, has proven efficacious in several disease models. However, these require highly specialized devices and skilled personnel, which precludes their use in mass vaccination especially in third world countries. Self-applicable, patch-based delivery technologies using DNA nanoparticles can be significantly less expensive, yet highly effective strategy for cutaneous vaccination. This proposal describes plans to gather preliminary data on the efficacy of plasmid DNA and CpG oligos delivered via polymer-nanoparticles and chemical enhancers, on skin penetration and subsequent generation of immune response against encoded antigens. The specific aims are: (a) synthesis of biodegradable nanoparticles of different size and surface charge with plasmid DNA and CpG oligos adsorbed on the surface and formulating them with various skin penetration agents, (b) studying the efficacy of particle transport and dendritic cell uptake following delivery with chemical enhancers and iontophoresis and (c) immune response against the encoded antigen following cutaneous application with a patch. These studies will enable us to identify the critical parameters for cutaneous particle delivery and further optimize and develop a simple, needle-free, skin-patch type system for DNA immunization in specific disease models.