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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 在我们NIH资助的研究项目IR 03 AI 082316“生殖支原体输卵管感染模型”中，我们旨在评估生殖支原体的生长、持续性和对MG的免疫应答。nemestrina输卵管袋模型。 在该模型中，雌性灵长类动物通过手术将自体输卵管袋植入腹壁。 三周后，用MG（或PBS对照）接种囊袋并每周收获以通过培养和qPCR评估活MG的存在。 用单个灵长类动物进行的初步实验产生了令人鼓舞的结果，在每个时间点的三个MG接种口袋中，在第1周的一个口袋中检测到MG DNA，在第2周的所有三个口袋中检测到MG DNA，在第3周没有检测到MG DNA，在第4周检测到一个口袋。 在第1周和第2周恢复了活性MG，但在其他时间点未恢复。 免疫印迹法测定，感染过程中诱导MG抗体，并与免疫显性粘附分子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联。 在中试实验之后，我们已经确定，改进的组织匀浆技术和接种到Vero细胞共培养物中大大提高了从灵长类动物组织中回收活MG。 此外，我们采用了一种改进的qPCR方法，该方法不仅更具特异性，而且更具成本效益，因为一次可以分析384个样品（与我们以前的方法的32个样品相比）。 计划在2011年年中将剩余的两只灵长类动物纳入该赠款。 这些研究将拓宽我们对女性上生殖道感染这种新兴病原体的理解。