SALPINGEAL INFECTION NODAL OF MYCOPLASMA GENITALIUM

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 在我们由美国国立卫生研究院资助的IR03AI082316，“生殖支原体输卵管感染模型”中，我们的目标是评估输卵管支原体口袋模型中MG的生长、持久性和免疫反应。在这个模型中，雌性灵长类动物被手术植入腹壁，并带有自体输卵管口袋。三周后，接种MG(或PBS对照)，每周收获一次，通过培养和qPCR评估存活MG的存在。对单一灵长类动物进行的初步实验取得了令人鼓舞的结果，在每个时间点接种MG的3个口袋中，1个在第1周检测到MG DNA，所有3个在第2周检测到MG DNA，在第3周没有检测到MG DNA，在第4周检测到1个MG。在第1周和第2周，MG活着，但在其他时间点没有检测到。经免疫印迹检测，MG抗体在感染过程中被诱导产生，并与免疫优势的粘附素分子mGpB和mGpC发生反应。最后，正在进行实验，以评估mgpB和mgpC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。在中试实验后，我们确定了改进的组织匀浆技术和接种到Vero细胞共培养中，大大提高了从灵长类组织中回收存活MG的能力。此外，我们采用了一种改进的定量聚合酶链式反应方法，这种方法不仅更特异，而且更具成本效益，一次可以分析384个样本(与我们以前方法的32个样本相比)。计划在2011年年中为这笔赠款中的其余两只灵长类动物接种疫苗。这些研究将扩大我们对女性上生殖道感染这种新出现的病原体的理解。

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.