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.
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