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

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性传播感染（STI），影响超过130 这是全球可预防失明的最常见原因。在联合 国家，性病引起的C。沙眼每年花费数十亿美元（古恩et al.1998）。因为 这种感染可以是无症状的，它可能多年不治疗，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。因此，我们认为， 迫切需要一种疫苗来预防由C.沙眼是明确的。尽管 尽管为研制衣原体疫苗作出了相当大的努力，但没有任何进展。研究表明 用主要衣原体外膜蛋白（MOMP）免疫可以诱导显著的保护作用 如果保留其天然结构，则可以在小鼠和非人类灵长类动物中抵抗感染和疾病。 然而，MOMP疫苗的配制是一个主要障碍，因为这种蛋白具有16个跨膜结构。 结构域，是40%疏水性的，组装为同源三聚体，并含有多个半胱氨酸， 二硫桥我们现在已经证明，我们可以产生多寡聚体，SDS抗性， 使用纳米脂蛋白颗粒（NLP）的MOMP的活性形式。此外，我们还可以首次 产生全长衣原体多态性膜蛋白（Pmps），另一组衣原体表面 已显示出强免疫原性的抗原。所编码的蛋白质可以被工程化为血清型蛋白质。 特异性地制备用于减轻沙眼衣原体相关影响的多血清型疫苗 致病性最后，我们已经开发了将联合收割机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性的疫苗制剂。这一初步突破是由 结合合成生物学方法和MOMP和PMP与载脂蛋白的无细胞共表达。 该提案的重点是进一步扩展MOMP介绍与相关PMP，所有PMP都在 NLP，以获得针对衣原体感染的高度保护性疫苗。

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.