PRIMATE MODEL FOR MYCOPLASMA GENITALIUM

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在我们的研究基金IR 21 AI 074898“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研究生殖支原体免疫发病机制的动物模型的能力。生殖道衣原体（MG）是一种新发现的生殖道病原体，其疾病谱和意义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、持续和诱导外宫颈、内宫颈和阴道中的先天性和适应性免疫应答，并上升到上生殖道引起输卵管炎的能力。此外，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，我们确定了从灵长类动物中回收的MG分离株在用于这种难养微生物体外生长的标准H肉汤中的生长受到抑制。随后，我们确定，这种抑制作用可以通过在H肉汤中稀释灵长类动物标本或通过将感染的标本与Vero细胞共培养来克服。 我们目前正在评估是否有其他支原体属。定殖这些猕猴的下生殖道介导这种抑制。 因此，我们已经调整了我们的灵长类动物方案，以常规地将灵长类动物标本包括到H琼脂平板（以独立地评估MG和其他支原体样微生物的存在）、Vero细胞培养物和H肉汤的系列稀释液中。 使用灵长类动物组织的重建实验（可通过WaNPRC尸检程序获得），我们还开发了改善灵长类动物组织均质化的方法，以最大限度地恢复活MG。 我们还表明，MG基因序列变异与表面蛋白的抗原变异一致，发生在灵长类动物感染，但不是在同一接种物的MG同时培养在体外，支持MG在体内的持久性的抗原变异的作用。

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.