TRANSCUTANEOUS IMMUNIZATION FOR DNA

本提案的目的是评估通过经皮免疫技术（TCI）递送DNA疫苗的潜力。结合DNA疫苗和TCI技术的潜在优势与DNA疫苗诱导细胞毒性T淋巴细胞的产生和识别构象决定因子的体液反应的能力有关。由于初步数据显示病毒颗粒大小的抗原在这种方法中具有免疫原性，因此通过TCI进行DNA疫苗接种成功的可能性很大。我们将研究递送DNA疫苗载体的可行性，这些载体被浓缩成小的、抗核酸酶的颗粒，或包装成无包膜的病毒衣壳。使用绿色荧光蛋白表达作为读数，将开发条件，以优化经皮递送聚乙烯亚胺浓缩DNA到表皮树突状细胞。还将研究各种TCI佐剂对递送的影响。为了优化通过DNA TCI诱导的体液和CTL反应，将采用编码流感病毒核蛋白的载体。最后，我们还将把DNA疫苗封装到sv40o病毒衣壳外壳中，因为这种颗粒在体内和体外均能保护DNA免受核酸酶的附着，并有效地将质粒递送到哺乳动物细胞核中。拟议的商业应用：经皮破伤风增强剂的开发可以降低与免疫相关的针扎和相关的针传疾病的风险，增加对免疫方案的遵守，通过消除对训练有素的人员和无菌设备的需要来改善疫苗接种的可及性，并减少成功免疫中与物理皮肤穿透相关的并发症。有效地递送用于疫苗接种的DNA，可以为一些假定DNA免疫是有利的疫苗打开使用这一策略的大门。

The objective of this proposal is to evaluate the potential for delivering DNA vaccines via transcutaneous immunization technology (TCI). The potential advantages for combining DNA vaccine and TCI technologies are related to the ability of DNA vaccines to elicit the production of cytotoxic T lymphocytes and humoral responses recognizing conformational determinants. The likelihood of success for DNA vaccination via TCI is significant since preliminary data show that antigens the size of virus particles are immunogenic in this approach. We will examine the feasibility of delivering DNA vaccine vectors that are condensed into small, nuclease resistant particles, or packaged into non- enveloped virus capsid coats. Using green fluorescent protein expression as a readout, conditions will be developed to optimize transcutaneous delivery of polyethylenimine-condensed DNA into epidermal dendritic cells. The effects of various TCI adjuvants on delivery will also be examined. To optimize the induction of humoral and CTL responses via DNA TCI, a vector encoding influenza virus nucleoprotein will be employed. Finally, we will also encapsulate DNA vaccines into SV4O virus capsid coats due to the demonstrated ability of such particles to protect DNA from nuclease attach and efficiently deliver plasmids into the nucleus of mammalian cells in vivo and in vitro. PROPOSED COMMERCIAL APPLICATIONS: The development of a Transcutaneous tetanus booster could decrease the risk of immunization- related needle sticks and associated needle-borne disease, increase compliance with immunization protocols, improve access to vaccination by eliminating the need for trained personnel and sterile equipment to achieve immunization and reduce the complications related to physical skin penetration in successful immunization. Efficient delivery of DNA for vaccination could open the use of this strategy for a number of vaccines where it is presumed that DNA immunization is advantageous.