SALPINGEAL INFECTION NODAL OF 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。 本研究的目的是：（1）评估M.生殖器在Macaca nemestrina输卵管袋模型中的作用;生殖器基因变异随着时间的推移在输卵管袋。 到目前为止，在华盛顿国家灵长类动物研究中心（WaNPRC）和传染病部门的资助下，对一只试验灵长类动物进行的实验成功实施。 25输卵管袋成功地形成了试点灵长类动物。 植入后三周，20个最佳形成的囊袋接种109个M基因组。生殖器菌株G37（16个袋）或单独的PBS（8个袋）。 然后用相同剂量的该菌株接种宫颈。 此后每周间隔（第7、14、23和28天），取出3个MG接种的囊袋和2个PBS接种的对照囊袋，用剃刀刀片切碎，悬浮于MTM中，并用于评估H肉汤中的生长（通过颜色从红色变为黄色）和基因组的增加（通过定量PCR测定）。 在每个时间点的三个MG接种的口袋中，在第7天的一个口袋中检测到MG基因组DNA，在第14天的所有三个口袋中检测到MG基因组DNA，在第23天没有检测到MG基因组DNA，在第28天的一个口袋中检测到MG基因组DNA。 在第7天和第14天，也从MG DNA阳性口袋中回收了活MG，但在第28天未回收。 PBS接种的对照袋中没有一个对活MG或MG基因组DNA呈阳性。 第7天在阴道中也检测到MG  接种后第28天，接种后第7 - 58天宫颈中，以及第14天宫颈细胞刷标本  接种后第58天（未采集第7天宫颈细胞刷标本）。 接种后23天，通过Western印迹法检测到MG的体液抗体，包括与表观大小与MgpB和MgpC（目标2中研究的蛋白质）一致的蛋白质反应的抗体。

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. The aims of this project are to (1) Assess the growth, persistence, cellular location, and the local and humoral immune response to M. genitalium in the Macaca nemestrina salpingeal pocket model, and (2) Evaluate M. genitalium gene variation over time in salpingeal pockets. Experiments performed to date on the one pilot primate were successfully implemented with funding from the Washington National Primate Research Center (WaNPRC) and the Division of Infectious Diseases. Twenty-five salpingeal pockets were successfully formed in the pilot primate. Three weeks subsequent to implantation, the 20 best formed pockets were inoculated either with 109 genomes of M. genitalium strain G37 (16 pockets) or PBS alone (8 pockets). The cervix was then inoculated with the same dosage of this strain. At weekly intervals thereafter (days 7, 14, 23, and 28), three MG-inoculated pockets and two PBS inoculated control pockets were removed, minced with razor blades, suspended in MTM, and used to assess growth in H broth (by a color change from red to yellow) and an increase in genomes (determined by quantitative PCR). Among the three MG-inoculated pockets per time point, MG genomic DNA was detected in one at d7, all three at d14, none at d23, and one at d28. Viable MG was also recovered from the MG DNA positive pockets at d7 and d14, but not at d28. None of the PBS-inoculated control pockets were positive for viable MG or MG genomic DNA. MG was also detected in the vagina d7  d28 post inoculation, in the cervix d7-58 post inoculation, and from a cervical cytobrush specimen d14  d58 post inoculation (a d7 cervical cytobrush specimen was not collected). Humoral antibodies to MG were detected 23d post-inoculation by Western blot, including those reacting with proteins with an apparent size consistent with MgpB and MgpC, the proteins to be studied in Aim 2.