Outer membrane vesicle vaccine against gonococcal and meningcoccal disease

淋病是第二种最常见的细菌性传播感染-最常见的是 衣原体，通常与淋病混合感染。全球约有8000万新淋病病例 每年一次。2017年，美国报告的病例超过55.5万例。女性淋病的严重后遗症 包括不孕不育、宫外孕和慢性盆腔疼痛。病原体淋病奈瑟菌(Ng) 临床上使用的几乎所有抗生素都对淋病产生了抗药性。对头孢曲松和头孢曲松的耐药性 阿奇霉素--推荐的一线治疗药物--预示着一个无法治愈的淋病时代。美国疾病控制与预防中心 已将Ng列为威胁级别为“紧急”的微生物。一种安全有效疫苗的研制 预防淋病是公共卫生的优先事项。我们的团队已经证明了一种名为 针对广泛表达的Ng脂低聚糖(LOS)表位(2C7表位)的2C7具有杀菌作用。一个 2C7表位的多肽模拟(模拟位)，当被配置为聚合物(多抗原肽)时，引起 杀菌单抗和减轻Ng在小鼠体内的阴道定植。受到最近的回顾的鼓舞 由洗涤剂提取的外膜囊泡组成的B群脑膜炎双球菌疫苗的观察 旨在遏制新西兰脑膜炎双球菌流行的(MeNZB)也减少了淋球菌病的发病率 疾病减少31%，我们将把淋球菌2C7表位整合到脑膜炎球菌外膜囊泡中 同时针对脑膜炎双球菌和淋球菌。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。除了经济上的优势，一个“泛欧-- Neisseria的疫苗将有助于解决已经排得满满的疫苗接种计划，并可能减轻耻辱 与防止性传播感染的疫苗有关。在目标1中，我们将开发脑膜炎双球菌天然外膜 囊泡(NOMV)，含有显示2C7表位的LO。脑膜炎球菌FH结合蛋白(FHBP)将 突变为取消与人FH的结合，从而诱导出比野生株更高的抗Nm杀菌抗体。 键入FHBP。我们还将删除RMP和H.8，这可能会分别引发针对Ng和Nm的阻断抗体。 诱导的抗体将测试补体依赖的杀菌活性对同源和 异源Ng菌株，并在我们开发的表达Ng的转基因小鼠中测试了与Ng的疗效 人类补体抑制剂FH和C4BP，以更好地模拟人类的补体环境。 我们将使用缺乏C9(溶菌孔形成的最后一步)的小鼠或缺乏中性粒细胞的小鼠 为了阐明候选疫苗的作用机制，也将确定保护相关因素。在……里面 目的2，我们将开发展示2C7的洗涤剂提取的外膜囊泡疫苗(Domvs)。 主要外膜蛋白PorA框架内的模拟表位多肽。卡介苗的免疫原性和免疫效果 该疫苗将按照目标1所述进行评估。

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.