Nanocarriers for transcutaneous delivery of vaccines

描述（由申请人提供）：在开发非注射（无针注射）疫苗替代传统疫苗方面已经付出了大量努力。与传统疫苗相比，非肠外注射疫苗具有许多潜在的优势，包括1)可能赋予粘膜和全身免疫，2)增加稳定性，3)延长保质期，4)不需要针头和需要经过专门培训的卫生保健专家来接种疫苗，5)可能降低成本。其中一种方法是经皮免疫（TCI），这是一种非侵入性、安全的将抗原直接递送到裸露皮肤上的方法。免疫是通过直接局部应用疫苗抗原来实现的。尽管TCI具有吸引力，但该技术受到大分子量疫苗抗原在完整皮肤上运输的相对低效率的限制。最近在经皮给药方面的创新，包括化学增强剂、电、超声和微针，证明了大分子通过皮肤渗透屏障（特别是角质层）运输的可行性。皮肤的外层是由紧密堆积的脂质分子组成的，这些致密的、结晶的脂质分子形成了防止水分流失和病原体进入的基本屏障。最近的证据表明，这种屏障可以通过结构合理的纳米粒子（纳米载体）来克服。该提案将比较三种不同的纳米载体（温度响应中空纳米球、纳米水凝胶和星形共聚物）结合模型疫苗抗原并通过角质层将抗原递送到表皮免疫应答细胞的能力。每种类型的纳米载体的专门组装赋予了角质层脂质通道内每种独特的性质和不同的相互作用。使用纳米载体递送疫苗是一种平台技术，适用于递送各种现有和潜在的疫苗。为了本提案的目的，我们将使用两种不同的蛋白质：1)牛血清白蛋白，它已被荧光标记以监测大分子抗原的结合和渗透；2)F1-V，一种来自鼠疫耶尔森氏菌的疫苗抗原，鼠疫的病原体，我们和其他人已经证明它可以防止毒力鼠疫杆菌的气溶胶攻击。拟议的研究将通过利用纳米载体的新特性，解决应用纳米技术递送疫苗的重要问题。这些研究的结果将广泛适用于各种疫苗和治疗方法，并将进一步突出纳米技术在科学和医学中的重要作用。

DESCRIPTION (provided by applicant): A great deal of effort has been directed towards developing nonparenteral (needle-free) alternatives to traditional vaccine delivery. Nonparenteral vaccines offer a number of potential advantages over traditional vaccines including 1) the potential to confer mucosal as well as systemic immunity, 2) increased stability, 3) increased shelf-life, 4) elimination of needles and the need for specially trained healthcare specialists to administer vaccines, and 5) potentially lower costs. One such approach, transcutaneous immunization (TCI), is a non-invasive, safe method of delivering antigens directly onto bare skin. Immunization is achieved by direct topical application of a vaccine antigen. Despite the attractiveness of TCI, the technology is limited by the relative inefficiency of transport of large molecular weight vaccine antigens across intact skin. Recent innovations in transdermal delivery of drugs, including chemical enhancers, electricity, ultrasound, and microneedles, demonstrate the feasibility of large-molecule transport through the skin's permeation barrier, specifically the stratum corneum. This outer layer of the skin is composed of tightly packed lipid molecules and the dense, crystalline arrangement of these lipids creates the essential barrier to prevent water loss and pathogen entry. Recent evidence has shown that this barrier can be overcome by properly structured nano-sized particles (nanocarriers). This proposal will compare three different nanocarriers (temperature-responsive hollow nanospheres, nanohydrogels, and star copolymers) for the ability to incorporate a model vaccine antigen and deliver that antigen through the stratum corneum to immunoresponsive cells in the epidermis. The specialized assembly of each type of nanocarrier gives each unique properties and different interactions within the lipid channels of the stratum corneum. The use of nanocarriers for vaccine delivery is a platform technology, applicable to delivery of a variety of existing and potential vaccines. For the purposes of this proposal, we will utilize two different proteins: 1) Bovine Serum Albumin that has been fluorescently labeled to monitor incorporation and permeation of a macromolecular antigen, and 2) F1-V, a vaccine antigen from Yersinia pestis, the causative agent of plague, which we and others have shown to protect against aerosol challenge with virulent Y. pestis. The proposed studies will address important questions in vaccine delivery by application of nanotechnology through the exploitation of the novel properties of nanocarriers. The findings of these studies will be broadly applicable to a variety of vaccines and therapeutics and will further highlight the important role of nanotechnology in science and medicine.