Induction of cross C. trachomatis serovar protection utilizing a polyvalent nanoparticle vaccine.

摘要 沙眼衣原体(CT)是全世界最常见的性传播细菌 病原体。在卫生条件差的国家，眼部感染CT可导致沙眼，最常见的 世界范围内可预防失明的常见原因。基于对小鼠的保护和血清学研究 分成15个主要血清型，其中8个占大多数性传播感染(STI)。在……里面 无论男性还是女性，大多数生殖器CT感染都是无症状的，因此没有得到治疗。即使是在 有症状的病例，除非及时和正确地实施治疗，否则长期后遗症包括 可发生盆腔炎、慢性腹痛、宫外孕和不孕症。因此， 接种疫苗是控制衣原体感染的最好方法。在这个项目中，我们想要检验假设 用Ct抗原配制的亚单位疫苗能引起强烈的免疫反应并诱导交叉血清型 保护雌性小鼠免受生殖器挑战。为了实现这一目标，我们将在C3H/HEN中进行测试 雌性小鼠，Ct主要外膜蛋白(MOMP)和多态疫苗 膜蛋白(PPM)。来自Ct高级血清的MOMP、MOMP多肽、MOMP DNA和RNA将是 使用基于纳米支架的纳米脂蛋白颗粒(NLP)配制和交付。通过以下方式进行免疫 本土的MOMP将帮助指导这些疫苗结构的生产。加强对生殖器的保护 在肠道，小鼠将通过使用佐剂的粘膜和全身途径接种疫苗。接种疫苗后，粘膜 系统的细胞和体液免疫反应将被确定，并与保护相关。 接种疫苗的动物将受到最具临床相关性的8种Ct血清型的挑战。 对感染的保护将通过确定阴道培养阳性的小鼠的数量来评估， 阴道培养阳性的总数、小鼠阴道脱落的时间和阴道脱落的严重程度。 为了确定对长期后遗症的保护，接种疫苗和挑战的雌性小鼠随后将 被关在笼子里的是已证实的雄性繁育小鼠和生育力和上生殖器病理，包括输卵管积水 编队，将进行评估。如果我们不能仅通过接种疫苗获得强大的、广泛的跨血清型保护 对于MOMP，我们将使用NLP测试含有MOMP和ppm C、G或H的多价疫苗。至 在接种疫苗后，确定每个抗原的中性、添加、协同或拮抗作用， 将描述对每个抗原的免疫反应，并将数据与保护相关联。我们的 目标是设计一种安全的基于纳米支架的亚单位疫苗，保护小鼠免受生殖器挑战 感染人类生殖道的Ct血清型。在这个项目结束时，保护性疫苗构建了 将为扩大生产和人类实施做好准备。一种针对CT的有效疫苗将会有 对世界范围内的健康和社会经济产生广泛影响。

ABSTRACT Throughout the world Chlamydia trachomatis (Ct) is the most common sexually transmitted bacterial pathogen. In countries with poor sanitary conditions ocular infections with Ct can result in trachoma, the most frequent cause of preventable blindness worldwide. Based on protection and serological studies in mice Ct is classified into 15 major serovars of which eight account for most of the sexually transmitted infections (STI). In men and women, the majority of genital Ct infections are asymptomatic and therefore not treated. Even in symptomatic cases unless therapy is implemented in a timely and correct manner long-term sequelae including pelvic inflammatory disease (PID), chronic abdominal pain, ectopic pregnancy and infertility can occur. Thus, vaccination is the best approach to control chlamydial infections. In this Project we want to test the hypothesis that a subunit vaccine formulated with Ct antigens elicits robust immune responses and induces cross-serovar protection in female mice against a genital challenge. To accomplish this goal we are going to test, in C3H/HeN female mice, vaccines formulated with the Ct major outer membrane protein (MOMP) and the polymorphic membrane proteins (Ppms). MOMP, MOMP peptides, MOMP DNA and RNA, from Ct senior serovars, will be formulated and delivered using Nano-Scaffold-Based Nano Lipoprotein Particles (NLP). Immunization with native MOMP will help guide the production of these vaccine constructs. To enhance protection in the genital tract, mice will be vaccinated by mucosal and systemic routes using adjuvants. Following vaccination, mucosal and systemic cellular and humoral immune responses will be determined and correlated with protection. Vaccinated animals will be challenged transcervically with the eight most clinically relevant Ct serovars. Protection against infection will be assessed by determining the number of mice with positive vaginal cultures, the total number of positive vaginal cultures, the length of time mice shed and the severity of vaginal shedding. To determine protection against long-term sequelae vaccinated and challenged female mice will subsequently be caged with proven male breeder mice and fertility and upper genital pathology, including hydrosalpinx formation, will be evaluated. If we cannot obtain a robust, broad cross-serovar protection by vaccinating only with MOMP, we will test a polyvalent vaccine with MOMP in combination with Ppm C, G or H using NLP. To ascertain the neutral, additive, synergistic, or antagonistic effects of each antigen, following vaccination, immune responses to each antigen will be characterized and the data will be correlated with protection. Our goal is to engineer a safe Nano-Scaffold-Based subunit vaccine that protects mice against a genital challenge with the Ct serovars that infect the human genital tract. At the end of this project protective vaccine constructs will be ready for scale-up production and human implementation. An efficacious vaccine against Ct will have a broad worldwide health and socioeconomic impact.