Abstract
Bacterial vaginosis (BV) is the most common vaginal infection worldwide and is associated with significant adverse sequelae. We have recently characterized vaginolysin (VLY), the human-specific cytotoxin produced by Gardnerella vaginalis and believed to play a critical role in the pathogenesis of BV and its associated morbidities. We hypothesize that novel antibody-based strategies may be useful for detection of VLY and for inhibition of its toxic effects on human cells. Using purified toxin as an immunogen, we generated polyclonal rabbit immune serum (IS) against VLY. A western blot of G. vaginalis lysate was probed with IS and a single band (57 kD) identified. Immunofluorescence techniques using IS detected VLY production by G. vaginalis. In addition, we have developed a sandwich ELISA assay capable of VLY quantification at ng/ml concentrations in the supernatant of growing G. vaginalis. To investigate the potential inhibitory role of IS on VLY-mediated cell lysis, we exposed human erythrocytes to VLY or VLY pretreated with IS and determined the percent hemolysis. Pretreatment with IS resulted in a significant reduction in VLY-mediated lysis. Similarly, both human cervical carcinoma cells and vaginal epithelial cells exhibited reduced cytolysis following exposure to VLY with IS compared to VLY alone. These results confirm that antibody-based techniques are an effective means of VLY detection. Furthermore, VLY antiserum functions as an inhibitor of VLY–CD59 interaction, mitigating cell lysis. These strategies may have a potential role in the diagnosis and treatment of BV.
Highlights
Bacterial vaginosis (BV) is the most common vaginal infection worldwide and is associated with significant adverse consequences including and preterm labor and delivery [1,2], post-partum endometritis [3], and an increased risk of HIV acquisition [4,5,6]
We developed novel antibodybased techniques for VLY detection
Immunofluorescent detection of VLY associated with whole G. vaginalis was detected microscopically using immune serum and fluorescently labeled anti-rabbit secondary antibodies (Figure 1B)
Summary
Bacterial vaginosis (BV) is the most common vaginal infection worldwide and is associated with significant adverse consequences including and preterm labor and delivery [1,2], post-partum endometritis [3], and an increased risk of HIV acquisition [4,5,6]. Reported prevalence rates range from 10–40% depending upon the population studied [7]. Suboptimal methods of diagnosis and a high percentage of asymptomatic patients make the true prevalence of BV difficult to ascertain. The pathogenesis of BV remains poorly understood. It is most commonly defined as a pathological state characterized by the loss of normal vaginal flora, Lactobacillus species, and overgrowth of other microbes including Gardnerella vaginalis, Bacteroides species, Mobiluncus species, and Mycoplasma hominis. Suggest a primary role for G. vaginalis as a specific and sexually transmitted etiological agent in BV, as was initially postulated by Gardner and Dukes in 1955 [8,9,10]
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