Heterologous expression in Tritrichomonas foetus of functional Trichomonas vaginalis AP65 adhesin
| Autor: | Alderete JF, Mundodi Vasanthakrishna, Kucknoor Ashwini S |
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| Rok vydání: | 2004 |
| Předmět: |
lcsh:QH426-470
Immunoblotting DNA Recombinant Protozoan Proteins Gene Expression Tritrichomonas foetus Cell Fractionation Transfection Cell Adhesion Trichomonas vaginalis Animals Humans lcsh:QH573-671 reproductive and urinary physiology Cell Line Transformed lcsh:Cytology Cell Membrane Antibodies Monoclonal Epithelial Cells lcsh:Genetics Protein Transport Microscopy Fluorescence embryonic structures Vagina Female Cell Adhesion Molecules Plasmids Research Article |
| Zdroj: | BMC Molecular Biology BMC Molecular Biology, Vol 6, Iss 1, p 5 (2005) |
| ISSN: | 1471-2199 |
| Popis: | Background Trichomonosis, caused by Trichomonas vaginalis, is the number one, nonviral sexually transmitted infection that has adverse consequences for the health of women and children. The interaction of T. vaginalis with vaginal epithelial cells (VECs), a step preparatory to infection, is mediated in part by the prominent surface protein AP65. The bovine trichomonad, Tritrichomonas foetus, adheres poorly to human VECs. Thus, we established a transfection system for heterologous expression of the T. vaginalis AP65 in T. foetus, as an alternative approach to confirm adhesin function for this virulence factor. Results In this study, we show stable transfection and expression of the T. vaginalis ap65 gene in T. foetus from an episomal pBS-ap65-neo plasmid. Expression of the gene and protein was confirmed by RT-PCR and immunoblots, respectively. AP65 in transformed T. foetus bound to host cells. Specific mAbs revealed episomally-expressed AP65 targeted to the parasite surface and hydrogenosome organelles. Importantly, surface-expression of AP65 in T. foetus paralleled increased levels of adherence of transfected bovine trichomonads to human VECs. Conclusion The T. vaginalis AP65 adhesin was stably expressed in T. foetus, and the data obtained using this heterologous system strongly supports the role of AP65 as a prominent adhesin for T. vaginalis. In addition, the heterologous expression in T. foetus of a T. vaginalis gene offers an important, new approach for confirming and characterizing virulence factors. |
| Databáze: | OpenAIRE |
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