Outer membrane vesicle vaccine against gonococcal and meningcoccal disease

淋病是第二种最常见的细菌性性传播感染-最常见的是 衣原体，通常与淋病合并感染。全球约有8000万新发淋病病例 每年。2017年，美国报告了超过555，000例病例。女性淋病的严重后遗症 包括不孕症、宫外孕和慢性盆腔疼痛。淋病奈瑟氏菌（Ng），病原体 淋病，已成为耐几乎每一种抗生素在临床使用。对头孢曲松耐药， 阿奇霉素--推荐的一线治疗--预示着淋病无法治愈的时代的到来。疾控中心 已将Ng列为威胁级别为“紧急”的微生物。开发安全有效的疫苗 防治淋病是公共卫生的优先事项。我们的研究小组已经表明，一种名为 靶向广泛表达的Ng脂寡糖（LOS）表位（2C 7表位）的2C 7是杀菌的。一 2C 7表位的肽模拟物（模拟位），当配置为聚合物（“多抗原肽”）时， 在小鼠中，Ng具有杀菌Ab并减弱Ng阴道定殖。在最近的回顾展上， 观察到由去污剂提取的外膜囊泡组成的B群脑膜炎球菌疫苗 （MeNZB）旨在减少新西兰的脑膜炎球菌流行，也减少了淋球菌感染事件。 疾病的31%，我们将纳入淋球菌2C 7表位到脑膜炎球菌外膜囊泡 以脑膜炎球菌和淋球菌为目标。这种创新的方法是有利的，因为这两种疾病 在青少年和年轻人中非常普遍。除了经济优势外，一个“泛- 奈瑟氏菌的疫苗将有助于解决已经拥挤的疫苗接种计划，并可能减轻耻辱 与性传播感染疫苗有关。在目标1中，我们将开发脑膜炎球菌天然外膜 包含LOS的NOMV，其展示2C 7表位。脑膜炎球菌FH结合蛋白（FHbp）将 突变以消除人FH的结合，从而引发比野生型更高的针对Nm的杀菌性Ab。 FHbp型。我们还将删除Rmp和H.8，它们可能分别引起针对Ng和Nm的阻断性Ab。 将测试引发的Ab针对同源和非同源抗体的补体依赖性杀菌活性。 异源Ng菌株并在转基因小鼠中测试对Ng的功效，我们已经开发了表达 人补体抑制剂FH和C4 BP，以更好地模拟人体内的补体环境。 我们将使用缺乏C9（溶菌孔形成的最后一步）的小鼠或缺乏PMNs的小鼠 阐明候选疫苗的作用机制，这也将确定保护的相关因素。在 目的2，我们将开发展示2C 7的去污剂提取的外膜囊泡疫苗（dOMV 模拟表位肽的主要外膜蛋白，PorA的框架。免疫原性和效力 该疫苗将按照目标1所述进行评价。

Gonorrhea is the second most common bacterial sexually transmitted infection – the most common is chlamydia, which often coinfects with gonorrhea. About 80 million new cases of gonorrhea occur worldwide annually. Over 555,000 cases were reported in the U.S. in 2017. Serious sequelae of gonorrhea in women include infertility, ectopic pregnancy and chronic pelvic pain. Neisseria gonorrhoeae (Ng), the causative agent of gonorrhea, has become resistant to almost every antibiotic in clinical use. Resistance to ceftriaxone and azithromycin – the recommended first-line of treatment – portends an era of untreatable gonorrhea. The CDC has listed Ng as a microorganism with a threat level of “Urgent.” Development of a safe and effective vaccine against gonorrhea is a public health priority. Our group has shown that a monoclonal antibody (mAb) called 2C7 that targets a widely-expressed Ng lipooligosaccharide (LOS) epitope (the 2C7 epitope) is bactericidal. A peptide mimic (mimitope) of the 2C7 epitope, when configured as a polymer (a ‘multiantigen peptide’) elicits bactericidal Abs and attenuates Ng vaginal colonization in mice. Encouraged by the recent retrospective observation that a group B meningococcal vaccine composed of detergent-extracted outer membrane vesicles (MeNZB) designed to curtail a meningococcal epidemic in New Zealand also reduced incident gonococcal disease by 31%, we will incorporate the gonococcal 2C7 epitope into meningococcal outer membrane vesicles to target both meningococci and gonococci. This innovative approach is advantageous because both diseases are highly prevalent in adolescents and young adults. In addition to the economic advantages, a ‘pan- Neisseria’ vaccine would help address an already crowded vaccination schedule and may mitigate the stigma associated with vaccines against STIs. In Aim 1, we will develop meningococcal native outer membrane vesicles (NOMVs) containing LOS that displays the 2C7 epitope. Meningococcal FH-binding protein (FHbp) will be mutated to abrogate binding of human FH and thereby elicit higher bactericidal Ab against Nm than wild- type FHbp. We will also delete Rmp and H.8, which may elicit blocking Ab against Ng and Nm, respectively. Elicited Abs will be tested for complement-dependent bactericidal activity against homologous and heterologous Ng strains and tested for efficacy versus Ng in transgenic mice we have developed that express the human complement inhibitors, FH and C4BP, to better simulate the complement environment in humans. We will use mice deficient in C9 (the last step in formation of the bacteriolytic pore) or mice depleted of PMNs to elucidate the mechanism of action of the candidate vaccine, which will also define correlates of protection. In Aim 2, we will develop detergent-extracted outer membrane vesicle vaccines (dOMVs) that display the 2C7 mimitope peptide in the framework of the major outer membrane protein, PorA. Immunogenicity and efficacy of this vaccine will be evaluated as described in Aim 1.