Cutaneous DNA immunization with polymer-nanoparticles

描述（由申请人提供）：基于DNA的疫苗具有在各种疾病中高度有效的潜力。它们也更便宜，易于大规模生产，并且可能比目前可用的疫苗更安全。 尽管最近在各种动物模型中取得了成功，但人类基因免疫的预防和治疗成功还远未实现，并且等待开发患者依从的、有效的、安全的和实用的基因递送系统。通过皮肤的无针免疫是目前递送方案的极好替代方案，因为（a）皮肤是天然设计用于免疫监视和调节的抗原呈递细胞的丰富环境，和（B）它是针注射的更安全和更符合患者的替代方案。使用高压介导的生物射弹或粉末递送的皮肤免疫的最新发展已经证明在几种疾病模型中有效。然而，这些需要高度专业化的设备和熟练的人员，这排除了它们在大规模疫苗接种中的使用，特别是在第三世界国家。使用DNA纳米颗粒的自我适用的、基于贴片的递送技术可以是显著更便宜的，但对于皮肤疫苗接种是非常有效的策略。该提案描述了收集关于通过聚合物纳米颗粒和化学增强剂递送的质粒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应答的功效的初步数据的计划。具体目标是：（a）用吸附在表面上的质粒DNA和CpG寡核苷酸合成不同尺寸和表面电荷的可生物降解的纳米颗粒，并将它们与各种皮肤渗透剂配制，（B）研究用化学增强剂和离子电渗法递送后颗粒转运和树突细胞摄取的功效，和（c）用贴剂皮肤施用后针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的、无针的、皮肤贴片型系统，用于特定疾病模型中的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.