MOMP-based recombinant antigens for a Chlamydia vaccine

用于衣原体疫苗的基于 MOMP 的重组抗原

• 批准号: 10471957
• 负责人: Paola Massari
• 金额: $ 24.11万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-19 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10471957
• 关键词: Acute; Address; Adjuvant; Amino Acid Sequence; Antibiotics; Antibodies; Antibody Avidity; Antibody titer measurement; Antigens; B-Lymphocytes; Blindness; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Carrier Proteins; Cells; Cellular Immunity; Centers for Disease Control and Prevention (U.S.); Cervicitis; Chlamydia; Chlamydia Infections; Chlamydia muridarum; Chlamydia trachomatis; Chromatography; Circular Dichroism Spectroscopy; Clinical; Development; Disease; Ectopic Pregnancy; Engineering; Epitopes; Escherichia coli; Eye Infections; Female; Fertility; Goals; Gonorrhea; HIV; Human; Human Papillomavirus; Hypersensitivity; Immune; Immune response; Immunity; Immunize; Immunoglobulin A; Immunoglobulin Constant Region; Immunoglobulin G; In Vitro; Inbred BALB C Mice; Infection; Infertility; Interferon Type II; Interleukin-1 beta; Interleukin-10; Interleukin-12; Interleukin-2; Interleukin-4; Interleukin-6; Length; Measures; Mediating; Modeling; Molecular Conformation; Mus; Natural Immunity; Nature; Neisseria lactamica; Organism; Pathology; Pelvic Inflammatory Disease; PorB porin; Production; Property; Public Health; Pulmonary Inflammation; Reaction; Recombinants; Reporting; Respiratory Tract Infections; Sexual Transmission; Sexually Transmitted Diseases; Spleen; Structure; Subgroup; Subunit Vaccines; Surface; Syphilis; T cell response; T-Cell Proliferation; T-Lymphocyte Epitopes; TNF gene; Testing; Trachoma; Tube; Urethritis; VDAC1 gene; Vaccine Antigen; Vaccinee; Vaccines; Vagina; Vaginal Douching; Whole Organism; Woman; aluminum sulfate; base; chlamydia vaccine; chronic abdominal pain; comorbidity; cross reactivity; cytokine; design; immunogenic; improved; long-term sequelae; lymph nodes; major outer membrane protein; male; men; mouse model; neutralizing antibody; novel; pathogenic bacteria; prevent; reproductive tract; respiratory challenge; response; scale up; vaccine candidate; vaccine development; vaccine trial

The overall goal of this project is to develop a vaccine against Chlamydia. Chlamydia trachomatis (Ct) is an obligate intracellular gram-negative organism and the most common cause of bacterial sexually transmitted infections (STIs) worldwide. Ct infections in women cause long-term sequelae i.e. pelvic inflammatory disease (PID), ectopic pregnancy and infertility, and are often associated with other STIs (HIV, HPV, gonorrhea and syphilis). Antibiotics do not prevent recurring infections. Development of a vaccine against Ct is a largely unmet need. Previous whole organisms-based vaccines only conferred limited protection from Ct ocular infections, with short-lived, serovar/subgroup-specific immunity and hypersensitivity reactions in some vaccinated individuals. The Chlamydia major outer membrane protein, MOMP, is a prime antigen candidate for a subunit vaccine. MOMP, a trimeric porin with 8 surface exposed loops, contains regions of sequence variability (variable domains, VDs) within loops 2, 3, 5 and 6 and regions of constant amino acid sequences (constant domains, CDs) flanking the VDs. The VDs and CDs contain B-cell and T-cell epitopes that elicit neutralizing antibodies and CD4+ T cell responses responsible for protective cellular immunity. The MOMP VDs are unique for each serovar and, if combined in a single vaccine, could induce broad coverage against all isolates. Development of a MOMP-based vaccine, however, presents barriers for production and use in humans. To fill the current gaps, in our previous R03 AI123885, we engineered novel recombinant chimeric antigens inserting the whole loops 2, 3, 5 and 6 of the C. muridarum (Cm) MOMP into a carrier protein structurally similar to MOMP, the PorB porin fromthe human commensal Neisseria lactamica. The recombinant PorB/VD antigens are suitable for recombinant production in a folded conformation, are immunogenic, induce cross-reactive antibodies to recombinant MOMP, native MOMP and whole Cm and are protective in a mouse model of Cm respiratory infection. Our approach addresses the shortcomings of MOMP production without hindering induction of functional, protective host immune responses to Chlamydia. The hypothesis is that equivalent Ct MOMP-based constructs, appropriately adjuvanted, will elicit protective immune responses against Ct infection and disease, which can be tested in a mouse model of transcervical Ct challenge. The Aims are 1) To design, express and purify Ct MOMP serovar F-based PorB/VD1- 3, PorB/VD1-4 and PorB/VD1-2-4 and examine structure properties of the recombinant constructs; 2) To define humoral and cellular immune responses to the PorB/VD constructs in mice using adjuvants that are suitable for use in humans without reactogenic responses and 3) To evaluate protection against Ct serovar F in the mouse model of Ct transcervical challenge. The results of our studies will fill a critical unmet need for improving public health worldwide and provide the groundwork for a broadly protective human Chlamydia vaccine. We will also define immune profiles induced by our vaccine that are associated with protection from infection and disease.

该项目的总体目标是开发一种针对衣原体的疫苗。沙眼衣原体（Ct）是一种 专性细胞内革兰氏阴性微生物和细菌性传播的最常见原因 全球性传染病（STIs）女性感染Ct会导致长期后遗症，即盆腔炎 (PID)，宫外孕和不孕症，并往往与其他性传播感染（艾滋病毒，人乳头瘤病毒，淋病和 梅毒）。抗生素不能预防复发性感染。开发针对Ct的疫苗在很大程度上是一个未满足的要求。 需要的以前的基于全生物体的疫苗仅赋予有限的保护免受Ct眼部感染， 在一些接种疫苗的个体中出现短暂的血清型/亚群特异性免疫和超敏反应。 主要衣原体外膜蛋白MOMP是亚单位疫苗的主要候选抗原。 MOMP是一种具有8个表面暴露环的三聚体孔蛋白，含有序列可变性区域（可变结构域， 环2、3、5和6内的恒定氨基酸序列（恒定结构域，CD）和侧翼的恒定氨基酸序列（恒定结构域，CD）的区域 的VDs。VD和CD含有B细胞和T细胞表位，其引发中和抗体和CD 4 + T细胞 负责保护性细胞免疫的反应。MOMP VD对于每个血清型是唯一的，如果 组合在单一疫苗中，可以诱导对所有分离株的广泛覆盖。开发基于MOMP的 然而，疫苗在人类中的生产和使用存在障碍。为了填补目前的空白，在我们以前的 R 03 AI 123885中，我们设计了新的重组嵌合抗原，其插入了 梭将鼠伤寒沙门氏菌（Cm）MOMP转化为与MOMP结构相似的载体蛋白，即来自人的PorB孔蛋白， 乳酸奈瑟氏球菌重组PorB/VD抗原适用于在大肠杆菌中重组生产。 折叠构象，具有免疫原性，诱导针对重组MOMP、天然MOMP 和整个Cm，并且在Cm呼吸道感染的小鼠模型中具有保护性。我们的方法解决了 MOMP生产的缺点是不妨碍功能性、保护性宿主免疫应答的诱导 衣原体感染假设是，适当佐剂化的等效的基于Ct MOMP的构建体将引起 针对Ct感染和疾病的保护性免疫应答，这可以在感染的小鼠模型中进行测试。 转铁蛋白Ct激发。目的：1）设计、表达和纯化Ct MOMP F型PorB/VD 1 - 1， 3、PorB/VD 1 -4和PorB/VD 1 -2-4并检查重组构建体的结构特性; 2）定义 使用适用于以下的佐剂在小鼠中对PorB/VD构建体的体液和细胞免疫应答： 在人类中使用而无反应原性应答，以及3）评价小鼠对Ct血清型F的保护作用 Ct转铁蛋白激发模型。我们的研究结果将填补一个关键的未满足的需要，改善公共 这将有助于全球健康，并为广泛保护人类衣原体疫苗奠定基础。我们还将 定义由我们的疫苗诱导的免疫特征，这些免疫特征与保护免受感染和疾病有关。