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

项目名称：纳米载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130多人 每年有数百万人失明，是全世界可预防的失明最常见的原因。在美国 在美国，沙眼衣原体引起的性传播感染每年造成数十亿美元的成本(Gunn等人)。1998年)。因为 这种感染可能是无症状的，可能会多年不治疗，并可能导致长期的后遗症， 包括盆腔炎、慢性腹痛、宫外孕和不孕症。因此， 公共卫生迫切需要疫苗来预防沙眼衣原体引起的疾病，这是显而易见的。尽管 在开发衣原体疫苗方面做出了相当大的努力，但没有一项即将到来。研究表明， 衣原体主要外膜蛋白(MOMP)免疫可产生显著的保护作用 在老鼠和非人类灵长类动物中都能抵抗感染和疾病，如果它的天然结构得到保护的话。 然而，由于该蛋白有16个跨膜蛋白，因此研制MOMP疫苗是一个主要障碍。 结构域，40%疏水性，以同源三聚体的形式组装，并含有多种半胱氨酸，可形成 二硫键。我们现在已经证明，我们可以生产出一种多寡聚、耐十二烷基硫酸钠和 使用纳米脂蛋白颗粒(NLP)的MOMP的活性形式。此外，我们还可以第一次 产生全长衣原体多形膜蛋白(PMPs)，这是衣原体表面的另一组 具有很强免疫原性的抗原。编码的蛋白质可以被改造成血清型 专门用于制造多血清疫苗以减轻沙眼衣原体相关的影响 致病性。最后，我们开发了将MOMP与佐剂相结合的方法，以提供独特的 在小鼠挑战研究中具有保护性的疫苗配方。这一最初的突破是通过 结合合成生物学方法和MOMP和PMPs与载脂蛋白的无细胞共表达。 这项提案的重点是进一步扩大最高管理计划的列报范围和相关的PMP，所有这些都是在 NLP，以实现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.