SALPINGEAL INFECTION NODAL OF MYCOPLASMA GENITALIUM

生殖支原体输卵管感染淋巴结

• 批准号: 8357618
• 负责人: PATRICIA A TOTTEN
• 金额: $ 15.66万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357618
• 关键词: Antibodies; Antigenic Variation; Autologous; Bacterial Adhesins; Coculture Techniques; DNA; Female; Funding; Grant; Growth; Harvest; Immune response; Implant; Infection; Methods; Modeling; Mycoplasma genitalium; National Center for Research Resources; Nodal; Primates; Principal Investigator; Reproductive Tract Infections; Research; Research Infrastructure; Resources; Sampling; Source; Techniques; Time; Tissues; United States National Institutes of Health; Vero Cells; Western Blotting; Woman; abdominal wall; cost; cost effective; improved; pathogen; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. In our NIH-funded grant, IR03AI082316, "Salpingeal Infection Model of Mycoplasma genitalium" we aim to assess the growth, persistence, and immune response to MG in the M. nemestrina salpingeal pocket model. In this model female primates are surgically implanted in the abdominal wall with autologous salpingeal pockets. Three weeks later the pockets are inoculated with MG (or PBS control) and harvested weekly to assess the presence of viable MG by culture and qPCR. Pilot experiments with a single primate yielded encouraging results in that among the three MG-inoculated pockets per time point, MG DNA was detected in one at Week 1, all three at Week 2, none at Week 3, and one at Week 4. Viable MG was recovered at Week 1 and Week 2 but not at the other time points. Antibodies to MG were induced during the infection as determined by Western blot and reacted with the immunodominant adhesin molecules MgpB and MgpC. Finally, experiments are underway to assess antigenic variation in MgpB and MgpC and correlate sequence changes to the presence of antibodies that recognize those sequences. Following the pilot experiment we have determined that improved tissue homogenization techniques and inoculation into Vero cell cocultures greatly improves the recover of viable MG from primate tissues. Furthermore we have adapted an improved qPCR method that is not only more specific but also more cost effective in that 384 samples can be analyzed at one (compared to 32 samples by our previous method). Plans are in place to inoculate the remaining two primates in this grant in mid-2011. These studies will broaden our understanding of upper reproductive tract infection with this emerging pathogen in women.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 在我们的NIH资助的IR03AI082316中，我们旨在评估Nemestrina Salpingeal M. nemestrina Salpingeal Pocket模型中对MG的生长，持久性和免疫反应。 在这种模型中，女性灵长类动物被手术植入带有自体盐袋的腹壁。 三周后，将口袋接种了MG（或PBS控制），并每周收获，以评估培养物和QPCR的可行MG的存在。 单个灵长类动物的飞行员实验产生了令人鼓舞的结果，因为在每个时间点的三个毫克接种口袋中，在第1周的第1周中检测到Mg DNA，第2周，第3周都没有，在第4周中没有一个。 MG在第1周和第2周回收，但在其他时间点没有回收。 通过蛋白质印迹确定，在感染过程中诱导了对MG的抗体，并与免疫粘着蛋白分子MGPB和MGPC反应。 最后，正在进行实验，以评估MGPB和MGPC中的抗原变异，并将序列变化与识别这些序列的抗体的存在相关。 在试点实验之后，我们确定了改进的组织均质化技术和接种到Vero细胞共培养物中，大大改善了从灵长类组织中恢复可行的毫克。 此外，我们已经改编了一种改进的QPCR方法，该方法不仅更具体，而且更具成本效益，因为可以通过一个方法分析384个样本（与以前的方法相比，32个样本）。 计划在2011年中期将剩余的两个灵长类动物接种计划。 这些研究将扩大我们对女性新兴病原体上生殖道感染的理解。