PRIMATE MODEL FOR MYCOPLASMA GENITALIUM

生殖支原体灵长类动物模型

• 批准号: 8172790
• 负责人: PATRICIA A TOTTEN
• 金额: $ 15.51万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172790
• 关键词: Agar; Animal Model; Antigenic Variation; Autopsy; Bacteria; Cell Culture Techniques; Chlamydia trachomatis; Coculture Techniques; Computer Retrieval of Information on Scientific Projects Database; Disease; Endocervix; Exocervix; Funding; Genes; Genital system; Grant; Growth; Immune response; In Vitro; Infection; Institution; Macaca; Macaca nemestrina; Mediating; Membrane Proteins; Methods; Modeling; Mycoplasma; Mycoplasma genitalium; Organism; Primates; Protocols documentation; Recovery; Research; Research Personnel; Resources; Role; Salpingitis; Source; Specimen; Tissues; United States National Institutes of Health; Vagina; Variant; Vero Cells; improved; in vivo; pathogen; programs; reconstruction; reproductive; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In our grant, IR21AI074898, "A Primate Model of Mycoplasma genitalium" we will assess the ability of pigtail macaques to serve as animal models to study the immunopathogenesis of M. genitalium (MG), a newly recognized reproductive tract pathogen with a disease spectrum and significance that appears to be very similar to Chlamydia trachomatis. Thus, we will assess the ability of cervically-inoculated MG to infect, persist, and induce an innate and adaptive immune response in the ectocervix, endocervix, and vagina, and ascend into upper reproductive tract to elicit salpingitis. Further we will evaluate the occurrence of MG gene variation during infection and its effect on survival of this bacterium. In our initial experiments, we determined that MG isolates recovered from primates were inhibited for growth in the standard H broth used for in vitro growth of this fastidious organism. Subsequently we determined that this inhibition could be overcome by dilution of the primate specimen in H broth or by co-culture of the infected specimen with Vero cells. We are currently evaluating whether other Mycoplasma spp. colonizing the lower genital tract of these macaques mediate this inhibition. As a result, we have adapted our primate protocols to routinely include inoculating primate specimens to H agar plates (to independently assess the presence of MG and other Mycoplasma-like organisms), into Vero cell cultures, and into serial dilutions of H broth. Using reconstruction experiments with primate tissue (available through the WaNPRC necropsy program), we have also developed methods to improve homogenization of primate tissues to maximize the recovery of viable MG. We have also demonstrated that MG gene sequence variation consistent with antigenic variation of surface proteins occurs during primate infection but not in the same inoculum of MG cultivated concurrently in vitro, supporting a role of antigenic variation in persistence of MG in vivo.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在我们的资助项目 IR21AI074898“生殖支原体的灵长类动物模型”中，我们将评估猪尾猕猴作为动物模型来研究生殖支原体 (MG) 的免疫发病机制的能力，这是一种新认识的生殖道病原体，具有疾病谱和其重要性似乎与沙眼衣原体非常相似。 因此，我们将评估宫颈接种的 MG 感染、持续、诱导子宫颈外、子宫颈内膜和阴道先天性和适应性免疫反应，并上升到上生殖道引发输卵管炎的能力。进一步我们将评估感染过程中MG基因变异的发生及其对该细菌存活的影响。在我们最初的实验中，我们确定从灵长类动物中回收的 MG 分离株在用于这种挑剔生物体外生长的标准 H 肉汤中生长受到抑制。随后我们确定，可以通过在 H 肉汤中稀释灵长类动物标本或通过将受感染的标本与 Vero 细胞共培养来克服这种抑制。 我们目前正在评估其他支原体是否存在。定居在这些猕猴的下生殖道介导了这种抑制作用。 因此，我们调整了灵长类动物实验方案，常规包括将灵长类动物标本接种到 H 琼脂平板上（以独立评估 MG 和其他支原体样生物体的存在）、Vero 细胞培养物以及 H 肉汤的连续稀释液。 通过灵长类动物组织的重建实验（可通过 WaNPRC 尸检计划获得），我们还开发了改善灵长类动物组织均质化的方法，以最大限度地恢复有活力的 MG。 我们还证明，与表面蛋白抗原变异一致的 MG 基因序列变异发生在灵长类动物感染期间，但在体外同时培养的 MG 同一接种物中却没有发生，这支持了抗原变异在 MG 体内持续存在中的作用。