Role of divalent cations, pH, cytoskeleton componentes and surface charge on the adhesion of Trichomonas vaginalis to a polystyrene substrate

Filho, Fernando Costa e Silva; Elias, Cezar Antonio; Souza, Wanderley de

doi:10.1590/s0074-02761987000300009

Cited by 13 publications

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“…Since both polystyrene and the cell surface of trichomonads are negatively charged (24,29,45), laminin could act by rendering the plastic surface less negative or even positive. Our results showed otherwise.…”

Section: Discussionmentioning

confidence: 99%

“…After 1 hr ofincubation nonadherent parasites were aspirated with a Pasteur pipette, and 2 ml of PBS containing 2.5% (vol/vol) glutaraldehyde was added. Adherent parasites were counted in an inverted microscope by using a transparent grid with 4-mm2 squares placed below the plastic dishes (29 (14). Laminin P1 fragment was prepared according to Rohde et al (31).…”

Section: Methodsmentioning

confidence: 99%

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Presence of laminin-binding proteins in trichomonads and their role in adhesion.

Filho

Souza

Lopes

1988

Proc. Natl. Acad. Sci. U.S.A.

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Adhesion is regarded as an important feature in the pathogenesis of various microorganisms. Ability to recognize extracellular matrix proteins, such as laminin or fibronectin, has been correlated with invasiveness. We report that laminin enhances the adhesion of the parasitic protozoa (MDCK) cell is one of the most studied epithelial cell lines; in vitro it shows some basic properties of a true epithelium such as (i) functional polarity (7), (ii) tight junctions (8), (iii) some dome formation (9), (iv) a highly active Na+,K+-ATPase located in the basolateral portion of the cell (10), and (v) a transepithelial electric potential (11). It is therefore considered a good model for studies on basic aspects of parasite-epithelium interaction (12, 13).Laminin is a noncollagenous glycoprotein of 800 kDa, found in all basement membranes (14), with the ability to promote cell adhesion, differentiation, shape, and motility (15, 16). These functions seem to be related to specific receptors present on the surface of normal (17-19) and tumor cells (20) as well as in some pathogenic trypanosomatids (21). Receptors have also been identified in various species of bacteria (22), including Staphylococcus aureus, in which a 52-kDa surface protein was characterized as laminin binding (23). In the present paper we report the presence of a laminin-binding protein on the surface of trichomonads and show that laminin mediates the attachment of these parasites to epithelial cells. MATERIALS AND METHODSParasites. The K strain of T. foetus and the Jt strain of T. vaginalis (24) were used in the present study. Both were cultivated in TYM (25) or medium 199 (GIBCO) supplemented with 10% bovine serum, either containing laminin or depleted of laminin by affinity chromatography on heparinSepharose (26), and incubated in an atmosphere of 95% air/5% CO2 at 370C. Only parasites in the exponential phase of growth were used. Parasites were harvested and washed twice by centrifugation at 1400 x g for 15 min in 0.01 M sodium phosphate buffer/0.15 M NaCl, pH 7.2 (PBS), and used for adhesion experiments.Cells. The MDCK cells were cultivated in medium 199 supplemented with 10% bovine serum at 370C in an atmosphere of 95% air/5% CO2 until they reached confluence. Cultures under study were washed once with PBS, and fresh culture medium with or without laminin was added. B16F10 mouse melanoma cells (a kind gift from Isaiah Fidler, Houston, TX) were similarly cultivated as previously described (27).Adhesion of Parasites to Plastic and MDCK Cell Surfaces.Parasites were washed twice with PBS, counted, and adjusted to 105 per ml, and 2 ml was poured into plastic Petri dishes coated or not with laminin (28). After 1 hr ofincubation nonadherent parasites were aspirated with a Pasteur pipette, and 2 ml of PBS containing 2.5% (vol/vol) glutaraldehyde was added. Adherent parasites were counted in an inverted microscope by using a transparent grid with 4-mm2 squares placed below the plastic dishes (29). For MDCK cells two types of experiments were performed...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Presence of laminin-binding proteins in trichomonads and their role in adhesion.

Filho

Souza

Lopes

1988

Proc. Natl. Acad. Sci. U.S.A.

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“…It has been demonstrated in several works that Trichomonas vaginalis can adhere to plastic surfaces in the presence of various agents and under different experimental conditions [24][25][26][27][28]. In addition, one group [28] demonstrated that the parasite can adhere to polystyrene, intrauterine devices, and vaginal rings.…”

Section: Monolayer Formation By T Vaginalismentioning

confidence: 99%

Trichomonas vaginalis: Monolayer and Cluster Formation—Ultrastructural Aspects Using High-Resolution Scanning Electron Microscopy

Ortiz,

Verdan,

Fortes

et al. 2023

Pathogens

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Trichomonas vaginalis is an extracellular protozoan parasite that causes human trichomoniasis, a sexually transmitted infection (STI) that affects approximately 270 million people worldwide. The phenomenon of T. vaginalis adhesion to inert substrates has been described in several reports. Still, very few studies on cluster formation have been conducted, and more detailed analyses of the contact regions between the parasites’ membranes in these aggregate formations have not been carried out. The present study aims to show that T. vaginalis forms a tight monolayer, similar to an epithelium, with parasites firmly adhered to the culture flask bottom by interdigitations and in the absence of host cells. In addition, we analyzed and compared the formation of the clusters, focusing on parasite aggregates that float in the culture flasks. We employed various imaging techniques, including high-resolution scanning electron microscopy, transmission electron microscopy, cytochemistry, TEM tomography, and dye injection. We analyzed whether the monolayer behaves as an epithelium, analyzing cell junctions, cell communication, and ultrastructural aspects, and concluded that monolayer formation differs from cluster formation in many aspects. The monolayers form strong adhesion, whereas the clusters have fragile attachments. We did not find fusion or the passage of molecules between neighbor-attached cells; there is no need for different strains to form filopodia, cytonemes, and extracellular vesicles during cluster and monolayer formation.

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Effect of a phorbol ester on basic surface properties of trichomonads

1992

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The effect of nanomolar concentrations of 12-O-tetradecanoilphorbol-13-acetate (TPA) on the cell surface of the urogenital parasitic protozoa Trichomonas vaginalis and Tritrichomonas foetus was evaluated by means of measurements of the parasites' surface tension, electrokinesis, lectin agglutination tests, and adhesion to inert substrates. TPA-treated parasites had their adhesion increased to both plastic and glass substrates. This was accompanied by increases in the parasites' net negative surface charge and also by changes in their surface tension. The lectin agglutination assays suggest that the increase in surface negativeness may be related in some extent to alterations in the oligosaccharide composition. Successive treatment of the microorganisms with TPA and sphingosine, a well-known competitive inhibitor of the phorbol ester active site, depressed the tendency of trichomonads to exhibit a phenotype of activated cells.

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Role of divalent cations, pH, cytoskeleton componentes and surface charge on the adhesion of Trichomonas vaginalis to a polystyrene substrate

Cited by 13 publications

References 0 publications

Presence of laminin-binding proteins in trichomonads and their role in adhesion.

Presence of laminin-binding proteins in trichomonads and their role in adhesion.

Trichomonas vaginalis: Monolayer and Cluster Formation—Ultrastructural Aspects Using High-Resolution Scanning Electron Microscopy

Effect of a phorbol ester on basic surface properties of trichomonads

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