BIOADHESIVE MICROSPHERES FOR ORAL DNA VACCINATION

DNA疫苗接种是一个重要的新概念，作为一种可行的和潜在的优于使用传统纯化抗原和减毒病毒的替代方案，它正迅速得到人们的接受。由于酶和粘膜的屏障，用于DNA疫苗应用的口服质粒在很大程度上仍未被探索。尽管如此，通过Peyer's补丁靶向肠道相关淋巴组织（GALT）的可能性为开发提供了令人信服的理由，该补丁是诱导全身和粘膜免疫的重要部位。利用颗粒摄取的生物现象，我们开发了一种热塑性，生物粘合剂，“纳米级”微球系统，能够在口服给药后将质粒DNA递送到Peyer's贴片中。本提案的技术目标侧重于建立DNA微胶囊化为基因口服递送提供途径的原理证明，特别是用于DNA疫苗应用。HIV DNA疫苗原型将使用一种专有技术——相反转纳米胶囊（PIN），被封装到聚富马酸-癸二酸（FA:SA）微球中。将包裹HIV DNA候选疫苗的微球喂给小鼠，并通过ELISA检测免疫应答（血清IgG、阴道/粪便IgA）。该平台的开发满足了对将DNA传递到替代组织的有效技术的需求，并提供了将DNA疫苗接种扩展到粘膜免疫至关重要的疾病的机会。建议的商业应用：口服途径是治疗性化合物给药的最简单途径。DNA疫苗口服递送系统的开发满足了将DNA递送到粘膜组织部位的有效技术的需要。

DNA vaccination is an important new concept which is rapidly gaining acceptance as both a viable and potentially superior alternative to the use of traditional purified antigen and attenuated viruses. Due to enzymatic and mucosal barriers, oral delivery of plasmids for DNA vaccine applications has remained largely unexplored. Nonetheless, the possibility of targeting the gut associated lymphoid tissue (GALT) via the Peyer's patches, an important site for induction of both systemic and mucosal immunity, provides a compelling rationale for development. Utilizing the biological phenomenon of particle uptake, we have developed a thermoplastic, bioadhesive, 'nano-sized' microsphere system capable of delivering plasmid DNA into the Peyer's patches following oral administration. The technical objectives of this proposal focus on establishing proof of principle that microencapsulation of DNA provides a pathway for oral delivery of genes, specifically for DNA vaccine applications. A HIV DNA vaccine prototype will be encapsulated into poly fumaric-co-sebacic acid (FA:SA) microspheres using a proprietary technique, phase inversion nanoencapsulation (PIN). Microspheres encapsulating HIV DNA vaccine candiate will be fed to mice and resulting immune response (serum IgG, vaginal/fecal IgA) will be measured by ELISA. The development of this platform fulfills the need for an effective technique to deliver DNA into alternative tissues and provides the opportunity to expand DNA vaccination to target diseases where mucosal immunity is critical. PROPOSED COMMERCIAL APPLICATION: The oral route is the simplest route for administration of therapeutic compounds. The development of an oral delivery system for DNA vaccination fulfills the need for an effective technique to deliver DNA into mucosal tissue sites.