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: 10288995
• 负责人: Paola Massari
• 金额: $ 21.85万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10288995
• 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; Individual; 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; Vaccinated; Vaccine Antigen; Vaccines; Vagina; Vaginal Douching; Whole Organism; Woman; aluminum sulfate; base; chlamydia vaccine; chronic abdominal pain; comorbidity; cross reactivity; cytokine; design; immunogenic; improved; lymph nodes; major outer membrane protein; male; men; mouse model; neutralizing antibody; novel; pathogenic bacteria; prevent; reproductive tract; respiratory; 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针对衣原体的疫苗。衣原体沙眼（CT）是 强制性细胞内革兰氏阴性生物和细菌性传播的最常见原因 全球感染（性传播感染）。女性的CT感染会引起长期后遗症，即骨盆炎性疾病 （PID），异位妊娠和不育，通常与其他性传播感染有关（HIV，HPV，淋病和 梅毒）。抗生素不能阻止反复感染。针对CT的疫苗开发在很大程度上是未修复的 需要。以前基于整个生物体的疫苗仅赋予有限的CT眼感染保护 一些接种疫苗的个体中短暂的，血清/亚组特异性免疫和超敏反应。 衣原体主要外膜蛋白MOMP是亚基疫苗的主要抗原候选者。 MOMP是一种具有8个表面暴露环的三聚体孔，包含序列可变性的区域（可变域， VDS）在环2、3、5和6中以及恒定氨基酸序列（恒定域，CD）的区域侧面 VDS。 VD和CD包含引起中和抗体和CD4+ T细胞的B细胞和T细胞表位 负责保护性细胞免疫的反应。 MOMP VD是每个血清的独特之处，如果 在单一疫苗中合并，可以引起所有分离株的广泛覆盖范围。基于MOMP的开发 然而，疫苗为人类的生产和使用壁垒提供了障碍。为了填补当前的空白，在我们以前的 R03 AI123885，我们设计了新颖的重组嵌合抗原，插入了整个环的2、3、5和6 Muridarum（CM）MOMP进入载体蛋白在结构上类似于MOMP，来自人类的porb porin 共生奈瑟氏菌。重组PORB/VD抗原适合重组生产 折叠构象，具有免疫原性，诱导重组MOMP的交叉反应抗体，本地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，而不会阻碍功能性，保护性宿主免疫反应的诱导 去衣原体。假设是，适当的佐剂将引起等效的基于CT MOMP的结构 防止CT感染和疾病的保护性免疫反应，可以在小鼠模型中进行测试 经胸CT挑战。目的是1）设计，表达和纯化CT MOMP血清基于F的PORB/VD1-- 3，PORB/VD1-4和PORB/VD1-2-4并检查重组构建体的结构特性； 2）定义 使用适合于小鼠的PORB/VD构建体的体液和细胞免疫反应，使用适合 在没有反应反应的人类中使用和3）评估小鼠中CT血清F的保护 CT经胸挑战的模型。我们的研究结果将满足改善公众的至关重要的需求 全球卫生健康，为广泛保护性的人衣原体疫苗提供基础。我们也会 定义由我们的疫苗诱导的免疫特征，与保护和疾病有关。