Analisi proteomica e funzionale del secreto diTrichomonas vaginalis

Trichomonas vaginalisis the causative agent of trichomoniasis, one of the most common sexually transmitted diseases in humans, recently associated with severe complications such as a significantly increased relative risk of developing cervical and prostate cancer. The mechanism by whichT. vaginalisexerts the pathogenesis is only partly characterized. It was demonstrated that several secreted molecules play a central role, but proteins implicated are never directly observed. It was observed that lysis is dependent on pH, with a peak of activity at pH 5.8, lost when pH is raised over 6.5.In our work, we optimised a protocol to obtain secreted proteins suitable for different proteomic approaches such as 1D and 2D electrophoresis. Our method allowed to identify in supernatants 114 and 202 proteins with heterogeneous functions obtained at pH 5.8 and 7.0, respectively.Through bioinformatic analysis and electron microscopy, we shown thatT. vaginalissecretes proteins via exosomes, a vesicular mediated mechanism that was not even reported inT. vaginalis.We observed several differences between secretomes at pH 5.8 and 7.0, in particular for classes of proteases.We identified an ortholog for human cytokine macrophage migration inhibitory factor that can be involved in the correlation with prostate cancer.The database constructed by this work can be a start point for studies in pathogenesis, drug discovery and development of vaccination strategies for the control of trichomoniasis.