Project 1. Nanolipoprotein-supported multi-subunit vaccine for Chlamydia trachomatis

项目1.纳米脂蛋白支持的沙眼衣原体多亚单位疫苗

• 批准号: 10458654
• 负责人: Matthew Adrian Coleman
• 金额: $ 53.06万
• 依托单位: LAWRENCE LIVERMORE NATIONAL SECURITY, LLC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-08 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458654
• 关键词: Adjuvant; Affect; Antigen Presentation; Antigens; Apolipoproteins; Bioinformatics; Biomedical Engineering; Blindness; Cells; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Codon Nucleotides; Collaborations; Complex; Coupled; Cryoelectron Microscopy; Cysteine; Disease; Disulfides; Ectopic Pregnancy; Engineering; Formulation; Genital; Genitalia; Hydrophobicity; Immunization; Immunology; In Vitro; Individual; Infection; Infertility; Length; Membrane Proteins; Methods; Mus; Pathogenesis; Pathogenicity; Pathology; Pelvic Inflammatory Disease; Persons; Plasmids; Production; Property; Protein Engineering; Proteins; Protocols documentation; Public Health; Recombinants; Research; Research Project Grants; Resistance; Safety; Sexually Transmitted Diseases; Solubility; Structure; Subunit Vaccines; Surface Antigens; Technology; Testing; Time; Translations; Transmembrane Domain; United States; Vaccines; Variant; Western Blotting; base; chronic abdominal pain; combinatorial; cost; immunogenicity; long-term sequelae; major outer membrane protein; mimetics; nano; nanolipoprotein particles; nanomembrane; nanoparticle; nonhuman primate; novel; pathogen; preservation; prevent; protein complex; protein expression; protein folding; protein purification; protein structure; safety testing; scaffold; scale up; statistics; synthetic biology; vaccine candidate; vaccine development; vaccine formulation

Project Title: Nanolipoprotein supported multi-subunit vaccine to Chlamydia trachomatis Project Abstract Chlamydia trachomatis is the most common bacterial sexually transmitted infection (STI), affecting over 130 million people every year, and is the most common cause of preventable blindness worldwide. In the United States, STIs caused by C. trachomatis account for billions of dollars in annual costs (Gunn et al. 1998). Because the infection can be asymptomatic, it may go untreated for years and can result in long-term sequelae, including pelvic inflammatory disease, chronic abdominal pain, ectopic pregnancy, and infertility. Therefore, the pressing public health need for a vaccine to prevent diseases caused by C. trachomatis is clear. Despite considerable efforts to develop a chlamydial vaccine, none have been forthcoming. Studies have shown that immunization with the Chlamydia major outer membrane protein (MOMP) can induce significant protection against infection and disease in both mice and non-human primates if its native structure is preserved. However, formulation of MOMP vaccines is a major hurdle given that this protein has 16 transmembrane domains, is 40% hydrophobic, assembles as a homotrimer, and contains multiple cysteines that can form disulfide bridges. We have now demonstrated that we can produce a multi-oligomeric, SDS-resistant, and active form of MOMP using nanolipoprotein particles (NLPs). Furthermore, for the first time we can also produce full-length Chlamydia polymorphic membrane proteins (Pmps), another group of chlamydia surface antigens that have shown potent immunogenicity. The encoded proteins can be engineered to be serovar specific to make multi-serovar vaccines for mitigating affects associated with Chlamydia trachomatis pathogenicity. Finally, we have developed methods to combine MOMP with adjuvants to provide a unique vaccine formulation that was protective in mouse challenge studies. This initial breakthrough was achieved by combining synthetic biology approaches and cell-free co-expression of MOMP and PMPs with apolipoproteins. This proposal is focused on further extending MOMP presentation with associated PMPs, all formulated within NLPs, to achieve a highly protective vaccine against Chlamydia infections.

项目名称：纳米脂蛋白支持的沙眼衣原体多亚单位疫苗 项目摘要 沙眼衣原体是最常见的细菌性传播感染 (STI)，影响超过 130 每年有数百万人失明，是全世界可预防性失明的最常见原因。在美国 在美国，由沙眼衣原体引起的性传播感染每年造成数十亿美元的损失（Gunn 等，1998）。因为 感染可能是无症状的，可能多年未得到治疗，并可能导致长期后遗症， 包括盆腔炎、慢性腹痛、宫外孕和不孕症。所以， 显然，公共卫生部门迫切需要一种疫苗来预防沙眼衣原体引起的疾病。尽管 为开发衣原体疫苗付出了巨大努力，但尚未取得进展。研究表明 使用主要衣原体外膜蛋白 (MOMP) 进行免疫可以产生显着的保护作用 如果其天然结构得以保留，则可以抵抗小鼠和非人类灵长类动物的感染和疾病。 然而，鉴于这种蛋白质有 16 次跨膜，MOMP 疫苗的配方是一个主要障碍 结构域，疏水性为 40%，组装为同源三聚体，并包含多个半胱氨酸，可形成 二硫桥。我们现在已经证明，我们可以生产多寡聚体、耐 SDS 且 使用纳米脂蛋白颗粒 (NLP) 的 MOMP 活性形式。此外，我们还第一次可以 产生全长衣原体多态性膜蛋白（Pmps），另一组衣原体表面 已显示出有效免疫原性的抗原。编码的蛋白质可以被设计成血清型 专门用于制造多血清疫苗，以减轻与沙眼衣原体相关的影响 致病性。最后，我们开发了将 MOMP 与佐剂结合的方法，以提供独特的 在小鼠攻击研究中具有保护作用的疫苗配方。这一初步突破是通过 将合成生物学方法与 MOMP 和 PMP 与载脂蛋白的无细胞共表达相结合。 该提案的重点是进一步扩展 MOMP 演示与相关的 PMP，所有内容均在 NLP，以获得针对衣原体感染的高度保护性疫苗。