SALPINGEAL INFECTION NODAL OF MYCOPLASMA GENITALIUM

生殖支原体输卵管感染淋巴结

• 批准号: 8172791
• 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/8172791
• 关键词: Antibodies; Cervical; Cervix Uteri; Color; Communicable Diseases; Computer Retrieval of Information on Scientific Projects Database; DNA; Funding; Genes; Genome; Genomics; Grant; Growth; Immune response; Infection; Institution; Location; Macaca nemestrina; Modeling; Mycoplasma genitalium; Nodal; Primates; Proteins; Research; Research Personnel; Resources; Source; Specimen; Time; United States National Institutes of Health; Vagina; Variant; Washington; Western Blotting; dosage; implantation; 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. 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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目的目的是（1）评估Macaca Nemestrina Salbingeal口袋模型中对生殖器支原体的生长，持久性，细胞位置以及局部和体液的免疫反应，以及（2）评估盐囊袋中时间内生殖器基因变异。 迄今为止，在一个飞行员灵长类动物上进行的实验通过华盛顿国家灵长类动物研究中心（WANPRC）和传染病司的资助成功实施。 飞行员灵长类动物成功形成了25个Salbingeal口袋。 植入后三周，将20个最佳的口袋与109个基因组G37（16口袋）或单独的PBS（8个口袋）接种。 然后将子宫颈用相同剂量的该菌株接种。 此后的每周间隔（第7、14、23和28天），将三个mg接种口袋和两个PBS接种的对照口袋去除，用剃须刀碎片切碎，悬挂在MTM中，用于评估H肉汤的生长（通过红色变为黄色）和基因组的增加（通过定量PCR确定）。 在每个时间点的三个mg接种口袋中，在D7处的一个，D14时，所有三个，d23中的一个，一个在D28中都检测到Mg基因组DNA。 还从D7和D14的Mg DNA阳性口袋中回收了可行的MG，但在D28处没有回收。 PBS接种对照口袋均未对可行的MG或MG基因组DNA阳性。 接种后接种后，在接种后宫颈D7 D28中也检测到MG，并在接种后接种后的宫颈细胞液样品D14 D58（未收集D7颈椎细胞液样品）。 通过蛋白质印迹检测到23D接种后的23D接种抗体，包括与具有明显大小的蛋白质反应的抗体，与MGPB和MGPC一致，在AIM 2中研究了蛋白质。