SALPINGEAL INFECTION NODAL OF MYCOPLASMA GENITALIUM

University Of Washington, Seattle WA

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Abstract

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 [unreadable]d28 post inoculation, in the cervix d7-58 post inoculation, and from a cervical cytobrush specimen d14 [unreadable]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.