Trichomonas vaginalis: characterization of a 39-kDa cysteine proteinase found in patient vaginal secretions.
Trichomonosis, a chronic sexually transmitted disease, remains a public health problem affecting yearly over 170 million people worldwide. This disease is caused by Trichomonas vaginalis, a protozoan flagellate rich in cysteine proteinases (CPs). Although CPs are involved in trichomonal cytopathogenicity, only few of them have been defined as virulence factors. In this study, we characterize a T. vaginalis 39-kDa proteinase (CP39) found in vaginal secretions from patients with trichomonosis. The CP39 proteinase bound to HeLa epithelial cells, vaginal epithelial cells (VECs), and human prostatic cancer cells (DU-145). CP39 did not bind to a human colon cancer (CaCo) cell line, suggesting tissue-specific binding. CP39 was found in six fresh trichomonad isolates tested. In two-dimensional gels, CP39 appeared as a single spot with a pI 4.5. CP39 is inhibited by E-64, stable at 50 degrees C, and active in a wide pH range (3.6-9.0), with an optimum pH at 7.0. In addition, CP39 degraded collagens I, III, IV, and V, human fibronectin, human hemoglobin, and human immunoglobulins A and G. Indirect immunofluorescence detected CP39 on the parasite surface with specific polyclonal antibody to purified CP39. Finally, CP39 was found to be immunogenic, as evidenced by detection on immunoblots with serum of patients with trichomonosis, but not control individuals. These data suggest that CP39 may play a role during trichomonal infection.
Hernández-Gutiérrez
Rodolfo
R
Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, Apdo. Postal 14-740, 07300, México, DF, Mexico.
Avila-González
Leticia
L
Ortega-López
Jaime
J
Cruz-Talonia
Fernando
F
Gómez-Gutierrez
Guillermo
G
Arroyo
Rossana
R
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D015153:Blotting, Western; D002448:Cell Adhesion; D003546:Cysteine Endopeptidases / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000201:enzymology / Q000469:parasitology; D013696:Temperature; D014247:Trichomonas Vaginitis / Q000201:enzymology* / Q000469:parasitology; D014246:Trichomonas vaginalis / Q000201:enzymology*; D014407:Tumor Cells, Cultured; D014621:Vagina / Q000201:enzymology* / Q000469:parasitology
D000913:Antibodies, Protozoan; D015853:Cysteine Proteinase Inhibitors; D007106:Immune Sera; D003546:Cysteine Endopeptidases