Trapping membrane proteins with adjuvant-carrying amphipols for vaccine formulati

描述（由申请人提供）：沙眼衣原体感染在世界范围内广泛存在。这种细菌病原体影响多个器官系统，产生急性症状和持续感染，可导致长期后遗症。如果及时实施抗生素治疗，可有效控制沙眼衣原体感染。不幸的是，许多病例无症状，其他病例治疗较晚，和/或治疗失败。研制沙眼疫苗的努力早在几十年前就开始了。最近，随着沙眼原体在性传播感染中的作用被发现，一些实验室集中精力研制一种预防生殖器感染的疫苗。我们的长期目标是设计出一种能够预防沙眼衣原体感染的疫苗。我们想要验证的假设是，用与两性相关的衣原体（APols）的天然主要外膜蛋白（MOMP）配制的疫苗可以保护雌性小鼠免受阴道内攻击。通常，膜蛋白（MPs）在使用洗涤剂的水溶液中保持可溶性。然而，洗涤剂往往使MPs不稳定，这可能导致保护性构象表位的丧失。apol是一种可以替代洗涤剂的新型高分子表面活性剂。它们已被证明特别有利于维持MP的结构、功能和稳定性。我们的初步实验表明，用pol捕获的MOMP配制的候选疫苗比基于洗涤剂的等效疫苗更有效。我们建议在这一观察的基础上，i)研究MOMP/APol复合物在不同类型疫苗制剂中的稳定性和结构，ii)测试携带佐剂的APol的保护作用。事实上，由于它们与MPs永久结合，APols可用于将佐剂与抗原MPs一起递送到靶细胞，这有望促进强大的全身和粘膜免疫反应。雌性小鼠将在阴道内受到感染，然后进行阴道培养。保护措施将根据阴道培养呈阳性的动物数量以及感染的严重程度和持续时间来确定。随后，将对这些小鼠进行交配，以确定其生育率。为了优化候选疫苗，我们将比较使用不同佐剂以不同方式与不同APols连接的不同疫苗接种方案所引起的免疫应答，从而调节佐剂在不同细胞室中的释放。总之，沙眼衣原体感染在发达国家和不发达国家都是一个主要的健康问题。本提案的目标是研制一种由携带佐剂的载脂蛋白捕获MOMP的疫苗。用疫苗降低这些感染的发病率和流行率将对全世界的健康产生重大影响。此外，基于双酚溶解膜蛋白的疫苗制剂的开发有可能影响广泛的病原体。

DESCRIPTION (provided by applicant): Chlamydia trachomatis infections are widespread throughout the World. This bacterial pathogen affects multiple organ systems producing acute symptomatology and persistent infections that can result in long-term sequelae. If implemented in a timely manner, antibiotic therapy is effective in controlling C. trachomatis infections. Unfortunately, many cases are asymptomatic, others are treated late, and/or are unsuccessfully managed. Efforts to produce a vaccine against trachoma were initiated decades ago. More recently, with the uncovering of the role of C. trachomatis in sexually transmitted infections, several laboratories have focused their efforts on producing a vaccine against genital infections. Our long-term goal is to engineer a vaccine that can protect against C. trachomatis infections. The hypothesis we want to test is that a vaccine formulated with the native major outer membrane protein (MOMP) of Chlamydia associated with amphipols (APols) can protect female mice against an intravaginal challenge. Generally, membrane proteins (MPs) are kept soluble in aqueous solutions using detergents. However, detergents tend to destabilize MPs, which can lead to the loss of protective conformational epitopes. APols are a novel type of polymeric surfactants that can substitute to detergents. They have been shown to be particularly favorable towards maintaining MP structure, function and stability. Our preliminary experiments indicate that candidate vaccines formulated with APol-trapped MOMP are more effective than their detergent-based equivalents. We propose to build on this observation by i) studying the stability and structure of MOMP/APol complexes in various types of vaccine formulations and ii) testing the protective effect of adjuvant-carrying APols. Indeed, as they associate permanently with MPs, APols can be used to deliver adjuvants to target cells concomitantly with antigenic MPs, which is expected to favor a strong systemic and mucosal immune response. Female mice will be challenged intravaginally and the course of the infection will be followed with vaginal cultures. Protection will be determined based on the number of animals with positive vaginal cultures and the severity and length of the infection. Subsequently, the mice will be mated to determine the fertility rates. In order to optimize the candidate vaccines, we will compare the immune response elicited by different vaccination protocols using various adjuvants linked in various ways to various APols, so as to modulate the release of the adjuvant in different cellular compartments. In conclusion, C. trachomatis infections are a major health problem in both developed and underdeveloped countries. The goal of this proposal is to formulate a vaccine with MOMP trapped by adjuvant-carrying APols. Decreasing the incidence and prevalence of these infections with a vaccine would have a major health impact worldwide. Furthermore, the development of vaccine formulations based on amphipol solubilized membrane proteins has the potential to impact a broad spectrum of pathogens.