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传播。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，可能导致长期后遗症。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，许多病例没有症状，其他病例治疗较晚，和/或治疗不成功。几十年前就开始了生产沙眼疫苗的努力。最近，随着沙眼衣原体在性传播感染中的作用的揭示，一些实验室已将重点放在生产针对生殖器感染的疫苗上。我们的长期目标是设计一种可以预防沙眼衣原体感染的疫苗。我们想要测试的假设是，用与两性醇 (APols) 相关的衣原体天然主要外膜蛋白 (MOMP) 配制的疫苗可以保护雌性小鼠免受阴道内攻击。通常，使用去污剂使膜蛋白 (MP) 保持可溶于水溶液。然而，去污剂往往会破坏 MP 的稳定性，从而导致保护性构象表位的丢失。 APols是一种新型聚合物表面活性剂，可以替代洗涤剂。它们已被证明特别有利于维持 MP 结构、功能和稳定性。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去污剂的等效疫苗更有效。我们建议在这一观察的基础上，通过 i) 研究 MOMP/APol 复合物在各种类型疫苗制剂中的稳定性和结构，以及 ii) 测试携带佐剂的 APol 的保护作用。事实上，由于 APols 与 MP 永久结合，因此可用于将佐剂与抗原 MP 一起递送至靶细胞，预计这将有利于强烈的全身和粘膜免疫反应。雌性小鼠将在阴道内受到攻击，并通过阴道培养物跟踪感染过程。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发的免疫反应，从而调节佐剂在不同细胞区室中的释放。总之，沙眼衣原体感染是发达国家和不发达国家的主要健康问题。该提案的目标是配制一种由携带佐剂的 APols 捕获 MOMP 的疫苗。通过疫苗降低这些感染的发生率和患病率将对全世界的健康产生重大影响。此外，基于两性醇溶解膜蛋白的疫苗制剂的开发有可能影响广泛的病原体。

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.