antibody-based detection and inhibition of vaginolysin, the gardnerella vaginalis cytolysin基于抗体的检测和抑制vaginolysin,加德纳菌属养狐场溶细胞素.pdfVIP

Antibody-Based Detection and Inhibition of Vaginolysin, the Gardnerella vaginalis Cytolysin 1 1 1 1,2 Tara M. Randis , Ritwij Kulkarni , Jorge L. Aguilar , Adam J. Ratner * 1 Department of Pediatrics, Columbia University, New York, New York, United States of America, 2 Department of Microbiology, Columbia University, New York, New York, United States of America 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 tec