Antigenic variation of Mycoplasma genitalium during persistent genital tract infection of pig-tailed macaques

猪尾猕猴生殖道持续感染过程中生殖支原体的抗原变异

• 批准号: 10350240
• 负责人: Gwendolyn Wood
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350240
• 关键词: Adherence; Alleles; Animal Model; Animals; Antibiotic Resistance; Antibodies; Antigenic Variation; Antigens; Antimicrobial Resistance; Appearance; Autopsy; Awareness; Azithromycin; Bacteria; Bacterial Adhesins; Cells; Cervical; Cervicitis; Cervix Uteri; Chlamydia trachomatis; Clinical; Development; Diagnostic; Diagnostic tests; Disease; Effector Cell; Enzyme-Linked Immunosorbent Assay; Exudate; FDA approved; Female; Genes; Goals; Histology; Immune; Immune Evasion; Immune response; Immunity; Immunologic Monitoring; In Situ; In Vitro; Individual; Infection; Infertility; Inflammation; Leucocytic infiltrate; Macaca nemestrina; Mammalian Oviducts; Modeling; Molecular; Moxifloxacin; Mycoplasma genitalium; National Institute of Allergy and Infectious Disease; Natural History; Nature; Neisseria gonorrhoeae; Pathogenesis; Pathology; Patients; Pelvic Inflammatory Disease; Phase; Population Study; Premature Birth; Prevalence; Primates; Process; Production; Protein Fragment; Proteins; Public Health; Recombinants; Recording of previous events; Research; Resistance; Role; Sequence Analysis; Serology test; Serum; Sexual Transmission; Sexually Transmitted Diseases; Specimen; Spontaneous abortion; Time; Tissues; Treatment Efficacy; Tumor-infiltrating immune cells; Urethritis; Variant; Woman; Work; animal model development; azithromycin resistance; biological specimen archives; chemokine; cytokine; detection assay; experimental study; genome sequencing; high risk; high risk population; improved; men; multiplex detection; pathogen; pathogenic bacteria; prevent; reproductive tract; resistant strain; whole genome

PROJECT SUMMARY Mycoplasma genitalium (MG), a sexually transmitted bacterial pathogen, is increasingly recognized as a significant public health concern. The prevalence of MG ranges from 1-4% in population-based studies to more than 20% in patients at high risk of acquiring sexually transmitted infections. The disease spectrum of MG is similar to Neisseria gonorrhoeae and Chlamydia trachomatis, and includes urethritis in men and cervicitis in women. Of particular concern, MG infection is associated with serious upper reproductive tract sequelae in women including pelvic inflammatory disease, infertility, preterm birth, and spontaneous abortion. Alarmingly, antimicrobial resistance in MG is increasing: 40-100% of strains are completely resistant to azithromycin and some infections are totally untreatable with US approved therapies. The recent FDA approval of two MG diagnostic tests will certainly increase public awareness of MG and demands for improved treatment. An animal model is urgently needed to understand the naturally history of MG infection including mechanisms of persistence and immune evasion, studies that are difficult in patients given the imperative to treat symptomatic infection. We have optimized our pig-tailed macaque model of persistent genital tract infection and now propose to use this model to study the role of antigenic variation in immune avoidance. Extending our previous work defining mechanisms of antigenic and phase variation of the immunodominant MgpB and MgpC adhesin proteins, we will determine if variation is required for persistence in the genital tract. First, using whole genome sequencing we will correlate the appearance of variants during 18 weeks of infection with the appearance of antibodies specific to MgpB and MgpC in three MG-infected primates and archived specimens from prior primate experiments. Second, the ability of the identical vs variant strain to re-infect animals that clear genital tract infection will be assessed in order to understand strain specific immunity. Third, a non-variable, “locked” MG strain that is unable to undergo antigenic variation will be constructed and characterized in vitro. Three primates will be inoculated cervically with a mixed inoculum of wild type and “locked” MG to compare the persistence and upper tract ascension of the two strains simultaneously over the 18 weeks of our model. All infected primates will undergo necropsy to examine the cervix, uterus, and Fallopian tubes for gross pathology and histology, and to assess the presence of MG in the upper reproductive tract. These experiments will not only determine if gene variation is required for persistence but will also provide additional observations in individual primates of the natural history of lower tract persistence, upper tract ascension, and immune response (including cytokines, cellular infiltrates and antibodies specific for conserved and variable MG antigens). These proposed experiments are highly significant in that they fill research gaps prioritized by NIAID-sponsored panels, namely the development of an animal model, the exploration of the role of MG in serious upper reproductive tract disease in women, and the development of an MG serologic test.

项目摘要 支原体生殖器（MG）是一种性传播细菌病原体，越来越被认为是一种 重要的公共卫生问题。 MG的患病率从基于人群的研究中的1-4％到更多 患有性传播感染的高风险患者的患者比20％。 MG的疾病频谱是 类似于淋病奈瑟氏菌和沙眼衣原体，包括男性尿道炎和宫颈炎 女性。特别关注的是，MG感染与严重的上层复制性续集有关 包括骨盆炎症性疾病，不育，早产和赞助堕胎在内的妇女。令人震惊的是 毫克抗菌素的耐药性正在增加：40-100％的菌株完全抗硫霉素和 有些感染对美国批准的疗法完全无法治疗。最近两毫克的FDA批准 诊断测试肯定会提高公众对MG的认识，并需要改善治疗方法。一个 迫切需要动物模型来了解MG感染的自然历史，包括 持久性和免疫避难所，在允许治疗症状的患者中很难进行研究 感染。我们已经优化了持续生殖道感染的猪尾猕猴模型 建议使用该模型研究抗原变异在免疫避免的作用。扩展我们的先前 免疫主导MGPB和MGPC粘附素的抗原和相变的工作定义机制 蛋白质，我们将确定生殖道持久性是否需要变异。首先，使用整个基因组 测序我们将在感染18周期间将变体的外观与外观相关联 来自先前的三个MG感染的主要和存档的标本中对MGPB和MGPC的抗体 灵长类动物实验。第二，相同与变体应变重新感染动物的能力清除生殖器 将评估区域感染，以了解特定的特异性免疫力。第三，一个不可变化的“锁定” 无法在体外构建和表征无法进行抗原变异的MG菌株。三 私人将与野生型混合接种物和“锁定”毫克的混合接种私人接种 仅在我们模型的18周内，这两种菌株的持久性和上缘提升。全部 受感染的原发性将经过尸检检查子宫颈，子宫和输卵管的严重病理 和组织学，并评估上部生殖道中MG的存在。这些实验不会 仅确定持久性是否需要基因变异，但还将在 下层持久性的自然历史的个人主要历史，上层升天和免疫 响应（包括细胞因子，细胞浸润和针对配置和可变MG的抗体 抗原）。这些提出的实验非常重要，因为它们填补了优先考虑的研究空白 NIAID赞助的面板，即动物模型的发展，MG在 女性的严重上层生殖道和MG血清学检查的发展。