Live Attenuated Recombinant Bacterial Delivery of Polysaccharide Vaccine Antigens

描述（由申请方提供）：细菌多糖是疫苗开发的极好靶点，因为它们是表面暴露的，并且通常是病原体上表达的最具保护性的抗原。然而，纯化的多糖通常是差的免疫原，特别是如果它们的免疫原性不通过与蛋白质缀合而增强。此类多糖-蛋白质缀合物疫苗通常生产起来复杂且昂贵，并且难以递送。为了证明利用多糖作为疫苗候选物的新方法的潜力，我们在鼠伤寒沙门氏菌的减毒株上表达了铜绿假单胞菌脂多糖（IPS）的O抗原部分。我们在小鼠模型系统中比较了不同的免疫途径以预防急性铜绿假单胞菌肺炎，发现鼻内接种疫苗比口服或腹膜内接种提供更好的保护。然而，我们不知道是否是抗体应答的位置、水平和/或类型负责这种保护。我们还表明，鼻内递送该疫苗可以保护小鼠在角膜创伤或烧伤后免受铜绿假单胞菌感染，但我们不知道在这些不同部位诱导免疫的最佳免疫途径，也不知道宿主因素如囊性纤维化对调节减毒重组S.鼠伤寒。同样不清楚的是，这种疫苗是否可以减轻与多糖-蛋白质缀合物疫苗相关的问题，包括亚型特异性和亚型抑制，以及排除酸不稳定的O抗原。本基金的具体目的是：（1）确定由减毒重组S.（1）确定基于鼠伤寒沙门氏菌O抗原的疫苗的最佳免疫途径，（2）确定在角膜损伤或烧伤后保护免受铜绿假单胞菌感染的最佳免疫途径，（3）确定疫苗是否可以保护CF小鼠免受铜绿假单胞菌肺部感染，和（4）测试另外的基于O抗原的疫苗的多糖表达、免疫原性和保护。总体而言，该项目的长期目标是了解针对这些不同感染的保护基础，这反过来将使我们能够制定针对这种重要的机会致病菌的疫苗开发的合理策略。我们预计，这些知识也将适用于其他病原体。

DESCRIPTION (provided by applicant): Bacterial polysaccharides are excellent targets for vaccine development as they are surface exposed and often the most protective antigens expressed on a pathogen. However, purified polysaccharides are generally poor immunogens, particularly if their immunogenicity is not enhanced by conjugation to a protein. Such polysaccharide-protein conjugate vaccines are often complicated and expensive to produce and difficult to deliver. In order to demonstrate the potential of a novel approach to utilize polysaccharides as vaccine candidates, we have expressed the O antigen portion of Pseudomonas aeruginosa lipopolysaccharide (IPS) on an attenuated strain of Salmonella typhimurium. We compared different routes of immunization to protect against acute P. aeruginosa pneumonia in a mouse model system and found that intranasal administration of the vaccine provides better protection than either oral or intraperitoneal delivery. However, we do not know whether it is the location, level, and/or type of antibody response that is responsible for this protection. We have also shown that intranasal delivery of this vaccine can protect mice against P. aeruginosa infections after corneal trauma or after burns, but do not know the optimal route of immunization to induce immunity at these different sites, nor do we know the effect of host factors such as cystic fibrosis on modulating the immunity induced by the attenuated recombinant S. typhimurium. Also unclear is whether such vaccines can alleviate problems associated with polysaccharide-protein conjugate vaccines including subtype specificity and subtype inhibition, and the exclusion of acid-liable O antigens. The specific aims of this grant are to (1) determine the basis for protective immunity to P. aeruginosa acute pneumonia mediated by the attenuated recombinant S. typhimurium O antigen-based vaccine, (2) determine the optimal route of immunization to protect against P. aeruginosa infections after corneal injury or burns, (3) determine whether the vaccine can protect CF mice from P. aeruginosa lung infections, and (4) test additional O antigen-based vaccines for polysaccharide expression, immunogenicity, and protection. Overall, the long-term goal of this project is to understand the basis of protection against these varied infections, which in turn will allow us to develop rational strategies for vaccine development against this important opportunistic pathogen. We anticipate that this knowledge will also be applicable to other pathogens.