Chemokine Profiling of Vaginal Epithelial Cells Exposed to Gardnerella Vaginalis
Gardnerella vaginalis (GV) is associated with bacterial vaginosis (BV), a dysbiosis that predisposes to preterm labor and increased susceptibility to sexually transmitted infections. The potential role of cytokines elaborated by the vaginal epithelium may impact BV symptoms and hence, is the topic of this research. We hypothesized that host-derived chemokine expression will correlate with Nugent (Gram stain species diversity) severity scores obtained from women with GV colonization. This was tested in a co-culture model of human vaginal epithelial cells (VECs) exposed to various clinically isolated strains of GV. These strains were obtained from ATCC/BEI as follows Two additional clinical isolates of GV (strains 14 and 15) with unknown Nugent scores were also obtained. To further characterize the strains, clade analysis was performed by PCR. VECs were grown in KSF media and GV strains were grown on V-agar, all incubated at 37º C in 5% CO2. For co-culture, 10 µl aliquots of each GV strain (obtained from an overnight culture) was inoculated into confluent cultures of VECs; uninfected VECs served as control. After 24 hours, culture medium was collected, centrifuged, and 500 µl aliquots of the supernatant was applied to a chemokine assay membrane (Proteome Profiler Human Cytokine Array R&D Systems chemokine profiling was performed according to the manufacturer’s protocol which tested for 36 analytes. Membrane profiling showed that 23 cytokines and chemokines were variously upregulated following exposure of VECs to GV, while others are down-regulated relative to the control. Continued investigation will attempt to correlate Nugent scores and placement of strains into clades 1-4. Our results indicate that co-culture with GV induces release of numerous chemokines from VECs. Future work will determine if these chemokines signal in an autocrine/paracrine manner and/or participate in the recruitment and activation of immune cells in response to GV colonization.
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"Chemokine Profiling of Vaginal Epithelial Cells Exposed to Gardnerella Vaginalis" (2018). MU-COM Research Day. 104.