Trapping membrane proteins with adjuvant-carrying amphipols for vaccine formulati

用携带佐剂的两性聚合物捕获膜蛋白用于疫苗配制

• 批准号: 8324510
• 负责人: Melanie J Cocco
• 金额: $ 33.66万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8324510
• 关键词: Acute; Adjuvant; Adult; Affect; Age; Animals; Antibiotic Therapy; Antigens; Bacteria; Binding; Birth; Cells; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Cholera Toxin Protomer B; Chronic; Complex; Country; Detergents; Drug Formulations; Ectopic Pregnancy; Engineering; Ensure; Epitopes; Escherichia coli; Eye; Female; Fertility Rates; Genital system; Goals; Gram-Negative Bacteria; Health; Immune; Immune response; Immunocompromised Host; Incidence; Individual; Infection; Infertility; Integral Membrane Protein; Laboratories; Lead; Length; Link; Lung; Malignant Neoplasms; Membrane; Membrane Proteins; Modeling; Molecular Conformation; Mothers; Mucosal Immune Responses; Mus; Newborn Infant; Organism; Parasites; Partner in relationship; Patients; Persons; Pneumonia; Polymers; Preparation; Prevalence; Property; Protein Family; Protein Region; Proteins; Protocols documentation; Reaction; Role; Severities; Sexually Transmitted Diseases; Solutions; Structure; Subunit Vaccines; Testing; Time; Trachoma; Vaccinated; Vaccination; Vaccines; Vagina; Virus; Water; Woman; Work; aqueous; base; body system; cell mediated immune response; design; genital infection; immunogenicity; improved; major outer membrane protein; novel; novel vaccines; pathogen; porin; protective effect; protein structure; protein structure function; research study; respiratory; response; surfactant; vaccine candidate; vaccine development; vaccine efficacy

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.

描述（由申请人提供）：沙眼衣原体感染在世界各地广泛传播。这种细菌病原体影响多个器官系统，产生急性炎症和持续感染，可导致长期后遗症。如果及时实施，抗生素治疗对控制C。沙眼感染不幸的是，许多病例没有症状，其他病例治疗较晚，和/或管理不成功。生产沙眼疫苗的努力始于几十年前。最近，随着C。在性传播感染中，几个实验室集中精力生产生殖器感染疫苗。我们的长期目标是设计一种可以预防C的疫苗。沙眼感染我们要检验的假设是，用与两性虫（APols）相关的衣原体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，使用洗涤剂使膜蛋白（MP）保持可溶于水溶液中。然而，去污剂倾向于使MP不稳定，这可导致保护性构象表位的丧失。APOL是一种新型的高分子表面活性剂，可以替代洗涤剂。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，与APol捕获MOMP配制的候选疫苗比其基于去污剂的等效物更有效。我们建议通过i）研究MOMP/APol复合物在各种类型的疫苗制剂中的稳定性和结构和ii）测试携带促排剂的APol的保护作用来建立这一观察结果。事实上，由于它们与MP永久结合，因此APOL可用于将佐剂与抗原性MP一起递送至靶细胞，预期这有利于强的全身和粘膜免疫应答。将对雌性小鼠进行阴道内攻毒，并使用阴道培养物跟踪感染过程。将根据阴道培养阳性的动物数量以及感染的严重程度和持续时间确定保护措施。随后，将小鼠交配以确定生育率。为了优化候选疫苗，我们将比较使用以各种方式连接到各种APOL的各种佐剂的不同疫苗接种方案引起的免疫应答，以便调节佐剂在不同细胞区室中的释放。结果表明，C.沙眼感染在发达国家和不发达国家都是一个主要的健康问题。该提议的目标是配制具有被携带促排剂的APOL捕获的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amphipol溶解的膜蛋白的疫苗制剂的开发具有影响广谱病原体的潜力。