PRIMATE MODEL FOR MYCOPLASMA GENITALIUM

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在我们的基金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研究生殖支原体(MG)的免疫病理机制的能力。因此，我们将评估宫颈接种的MG感染、持续并在宫颈外、宫颈内和阴道诱导先天性和获得性免疫反应，并上升到上生殖道引发输卵管炎的能力。此外，我们还将评估感染过程中MG基因变异的发生及其对该细菌生存的影响。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的体外生长的标准H肉汤中被抑制生长。随后，我们确定这种抑制可以通过在H肉汤中稀释灵长类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。在这些猕猴的下生殖道定居是这种抑制的中介。因此，我们修改了我们的灵长类实验方案，常规地包括将灵长类标本接种到H琼脂平板上(以独立评估MG和其他类似支原体的生物的存在)，接种到Vero细胞培养中，并接种到H肉汤的连续稀释中。利用灵长类组织的重建实验(可通过WaNPRC尸检计划获得)，我们还开发了改进灵长类组织均质化的方法，以最大限度地恢复存活的MG。我们还证明了在灵长类动物感染期间，MG基因序列发生了与表面蛋白抗原变异一致的变异，但在体外同时培养的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.