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Trichomonas vaginalis interactions with fibronectin and laminin

Department of Microbiology, The University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78284-7758, USA¹

Author for correspondence: John F. Alderete. Tel: +1 210 567 3940. Fax: +1 210 567 6612. e-mail: ALDERETE{at}UTHSCSA.EDU

The sexually transmitted protozoan Trichomonas vaginalis cytoadheres to vaginal epithelial cells and causes contact-dependent cytotoxicity which, when combined with the normal exfoliation process, leads to erosion of the epithelium, which may allow trichomonads into extracellular matrix and basement membrane sites. Therefore, the association of T. vaginalis with immobilized fibronectin (FN) and laminin (LM) on cover-slips was examined. Binding of live parasites to coated cover-slips was time- and parasite-density-dependent. Coincubation with an inhibitor of trichomonad cysteine proteinases resulted in an increased attachment of parasites to FN but had no effect on binding to LM, denoting that protease activity influenced optimal FN associations. Further, 20 h mid-exponential phase trichomonads placed in fresh culture medium for 3 h gave higher levels of binding to FN, suggesting that changes during growth in vitro to T. vaginalis organisms affect maximal levels of binding to FN. Extended incubation with substrates diminished the capacity of parasites to bind FN and LM. Treatment of live organisms with periodate reduced binding to LM but not FN, suggesting a role for carbohydrates. In addition, trypsinization of live parasites decreased numbers bound to both substrates. Placement of trypsinized parasites in medium for 2 h fully regenerated binding to FN but not LM. Incubation of trypsinized parasites with cycloheximide abrogated regeneration of attachment to FN, affirming a role for synthesized surface proteins in FN binding. Importantly, the T. vaginalis adhesin proteins that mediate cytoadherence, and iron, a factor that regulates adhesin synthesis, were not involved in FN and LM recognition. These results suggest a role for surface proteins and carbohydrates in trichomonal associations with FN and LM, respectively.

Keywords: colonization, fibronectin, laminin, pathogenesis, Trichomonas vaginalis

Abbreviations: BM, basement membrane; ECM, extracellular matrix; FN, fibronectin; LM, laminin; TLCK, N-p-tosyl-L-lysine chloromethyl ketone; VECs, vaginal epithelial cells

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