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: 8711230
• 负责人: Melanie J Cocco
• 金额: $ 34.1万
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8711230
• 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; 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; biophysical properties; 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。这种细菌病原体会影响产生急性症状学和持续感染的多个器官系统，这些系统可能导致后遗症。如果及时实施，抗生素疗法可有效控制沙眼感染。不幸的是，许多病例是无症状的，其他病例被迟到和/或没有成功管理。几十年前，开始了针对沙丘瘤的疫苗的努力。最近，随着沙眼梭菌在性传播感染中的作用，一些实验室将其努力集中在为生殖器感染的疫苗上产生疫苗。我们的长期目标是设计一种可以预防沙眼梭菌感染的疫苗。我们要检验的假设是，与埋有木果（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爆内挑战的侵害。通常，膜蛋白（MPS）使用洗涤剂溶于水溶液中。但是，洗涤剂倾向于破坏MP的稳定，这可能导致保护性构象表位的丧失。 Apols是一种可以代替洗涤剂的新型聚合物表面活性剂。它们已被证明对维持MP结构，功能和稳定性特别有利。我们的初步实验表明，用APOL捕获的MOMP配制的候选疫苗比基于洗涤剂的等效物更有效。我们建议通过i）研究在各种类型的疫苗制剂中研究MOMP/APOL复合物的稳定性和结构，ii）测试辅助辅助apols的保护作用。实际上，由于它们与MPS永久关联，因此可以使用APOL将佐剂与抗原MPS同时交付靶细胞，后者有望有利于强烈的全身性和粘膜免疫反应。雌性小鼠将受到静脉内的挑战，并随着阴道培养的方式感染过程。保护将根据具有阴道阳性培养的动物数量以及感染的严重程度和长度来确定。随后，将配合小鼠以确定生育率。为了优化候选疫苗，我们将使用以各种方式连接到各种APOL的各种辅助者来比较不同的疫苗接种方案引起的免疫反应，以调节在不同的细胞室内佐剂的释放。总之，在发达国家和发达国家中，沙眼梭菌感染是一个主要的健康问题。该提议的目的是制定一种被辅助载体捕获的MOMP的疫苗。通过疫苗降低这些感染的发病率和患病率将在全球范围内产生重大健康影响。此外，基于Amphipol溶解的膜蛋白的疫苗制剂的开发具有影响广泛的病原体的潜力。

项目成果

A Recombinant Chlamydia trachomatis MOMP Vaccine Elicits Cross-serogroup Protection in Mice Against Vaginal Shedding and Infertility.

重组沙眼衣原体 MOMP 疫苗可对小鼠产生跨血清群保护，防止阴道脱落和不孕。

• DOI: 10.1093/infdis/jiz438
• 发表时间: 2020
• 期刊: The Journal of infectious diseases
• 作者: Tifrea,DeliaF;Pal,Sukumar;delaMaza,LuisM
• 通讯作者: delaMaza,LuisM

Labeling and functionalizing amphipols for biological applications.

用于生物应用的两性聚合物的标记和功能化。

• DOI: 10.1007/s00232-014-9655-y
• 发表时间: 2014
• 期刊: The Journal of membrane biology
• 作者: LeBon,Christel;Popot,Jean-Luc;Giusti,Fabrice
• 通讯作者: Giusti,Fabrice

Amphipols for each season.

• DOI: 10.1007/s00232-014-9666-8
• 发表时间: 2014-10
• 期刊: JOURNAL OF MEMBRANE BIOLOGY
• 影响因子: 2.4
• 作者: Zoonens, Manuela;Popot, Jean-Luc
• 通讯作者: Popot, Jean-Luc

A step closer to membrane protein multiplexed nanoarrays using biotin-doped polypyrrole.

• DOI: 10.1021/nn406252h
• 发表时间: 2014-02-25
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Della Pia EA;Holm JV;Lloret N;Le Bon C;Popot JL;Zoonens M;Nygård J;Martinez KL
• 通讯作者: Martinez KL

Long-term stability of a vaccine formulated with the amphipol-trapped major outer membrane protein from Chlamydia trachomatis.

• DOI: 10.1007/s00232-014-9693-5
• 发表时间: 2014-10
• 期刊: JOURNAL OF MEMBRANE BIOLOGY
• 影响因子: 2.4
• 作者: Feinstein, H. Eric;Tifrea, Delia;Sun, Guifeng;Popot, Jean-Luc;de la Maza, Luis M.;Cocco, Melanie J.
• 通讯作者: Cocco, Melanie J.