Scanning electron microscopy in the investigation of the in vitro hemolytic activity of Trichomonas vaginalis

Rosset, Iveli; Tasca, Tiana; Tessele, Paola Martins; Carli, Geraldo A De

doi:10.1007/s00436-001-0555-6

Cited by 11 publications

(8 citation statements)

References 12 publications

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“…In the case of T. vaginalis , cysteine peptidases have been implicated in the virulence [35]–[37], the cytotoxicity [29], the adherence to host cells [35], [36], [38], [39] and the detachment of host cells [40]. Furthermore, their action was associated with nutrient acquisition [37], [41], hemolysis [42] and the evasion of the host immune response [43]–[45].…”

Section: Introductionmentioning

confidence: 99%

Cysteine Peptidases, Secreted by Trichomonas gallinae, Are Involved in the Cytopathogenic Effects on a Permanent Chicken Liver Cell Culture

et al. 2012

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Trichomonas gallinae, the aetiological agent of avian trichomonosis, was shown to secrete soluble factors involved in cytopathogenic effect on a permanent chicken liver (LMH) cell culture. The present study focused on the characterization of these molecules. The addition of specific peptidase inhibitors to the cell-free filtrate partially inhibited the monolayer destruction, which implied the presence of peptidases in the filtrate and their involvement in the cytopathogenic effect. One-dimensional substrate (gelatin) SDS-PAGE confirmed the proteolytic character of the filtrate by demonstrating the proteolytic activity within the molecular weight range from 38 to 110 kDa. In addition, the proteolytic activity was specifically inhibited by addition of TLCK and E-64 cysteine peptidase inhibitors implying their cysteine peptidase nature. Furthermore, variations in the intensity and the number of proteolytic bands were observed between cell-free filtrates of low and high passages of the same T. gallinae clonal culture. Two-dimensional substrate gel electrophoresis of concentrated T. gallinae cell-free filtrate identified at least six proteolytic spots. The mass spectrometric analysis of spots from 2-D gels identified the presence of at least two different Clan CA, family C1, cathepsin L-like cysteine peptidases in the cell-free filtrate of T. gallinae. In parallel, a PCR approach using degenerated primers based on the conserved amino acid sequence region of cysteine peptidases from Trichomonas vaginalis identified the coding sequences for four different Clan CA, family C1, cathepsin L-like cysteine peptidases. Finally, this is the first report analyzing molecules secreted by T. gallinae and demonstrating the ubiquity of peptidases secreted by this protozoon.

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Section: Introductionmentioning

confidence: 99%

Cysteine Peptidases, Secreted by Trichomonas gallinae, Are Involved in the Cytopathogenic Effects on a Permanent Chicken Liver Cell Culture

et al. 2012

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“…Surface proteins are the most extensively studied molecules in trichomonad adhesion, although much controversy remains in the field (2,10). Previous reports suggest that proteins that bind to extracellular matrix components (4,14,16), cysteine proteinases (6,39,42), erythrocyte-specific ligands (17,24,45), and adhesins (3,5) are found to be involved in attachment of parasite to cells. Carbohydrates on the surface of trichomonads have received much less attention, but several studies have shown their potential role in adhesion.…”

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confidence: 99%

Trichomonas vaginalis Lipophosphoglycan Mutants Have Reduced Adherence and Cytotoxicity to Human Ectocervical Cells

et al. 2005

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The extracellular human pathogen Trichomonas vaginalis is covered by a dense glycocalyx thought to play a role in host-parasite interactions. The main component of the glycocalyx is lipophosphoglycan (LPG), a polysaccharide anchored in the plasma membrane by inositol phosphoceramide. To study the role of LPG in trichomonads, we produced T. vaginalis LPG mutants by chemical mutagenesis and lectin selection and characterized them using morphological, biochemical, and functional assays. Two independently selected LPG mutants, with growth rates comparable to that of the wild-type (parent) strain, lost the ability to bind the lectins Ricinnus comunis agglutinin I (RCA120) and wheat germ agglutinin, indicating alterations in surface galactose and glucosamine residues. LPG isolated from mutants migrated faster than parent strain LPG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting the mutants had shorter LPG molecules. Dionex high-performance anion exchange chromatography with pulsed amperometric detection analyses revealed galactosamine, glucosamine, galactose, glucose, mannose/xylose, and rhamnose as the main monosaccharides of T. vaginalis parent strain LPG. LPG from both mutants showed a reduction of galactose and glucosamine, corresponding with the reduced size of their LPG molecules and inability to bind the lectins RCA120 and wheat germ agglutinin. Mutant parasites were defective in attachment to plastic, a characteristic associated with avirulent strains of T. vaginalis. Moreover, the mutants were less adherent and less cytotoxic to human vaginal ectocervical cells in vitro than the parental strain. Finally, while parent strain LPG could inhibit the attachment of parent strain parasites to vaginal cells, LPG from either mutant could not inhibit attachment. These combined results demonstrate that T. vaginalis adherence to host cells is LPG mediated and that an altered LPG leads to reduced adherence and cytotoxicity of this parasite.

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“…the causative agent of trichomoniasis, a serious sexually transmitted infection affecting over 180 million people worldwide, proteases have been implicated as virulence factors (2)(3)(4)(5)(6)(7)(8), as adherence factors (4 -9), as a cell-detaching factor (10), in nutrient acquisition (2,11), and hemolysis (12,13). It has also been suggested that CPs contribute to pathogenesis when released into the host mucosal surface (2,14) and that they may have roles in evasion of the host immune response (14 -18).…”

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confidence: 99%

Identification of Trichomonas vaginalis Cysteine Proteases That Induce Apoptosis in Human Vaginal Epithelial Cells

Sommer

Costello

Hayes³

et al. 2005

Journal of Biological Chemistry

108

114

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Scanning electron microscopy in the investigation of the in vitro hemolytic activity of Trichomonas vaginalis

Cited by 11 publications

References 12 publications

Cysteine Peptidases, Secreted by Trichomonas gallinae, Are Involved in the Cytopathogenic Effects on a Permanent Chicken Liver Cell Culture

Cysteine Peptidases, Secreted by Trichomonas gallinae, Are Involved in the Cytopathogenic Effects on a Permanent Chicken Liver Cell Culture

Trichomonas vaginalis Lipophosphoglycan Mutants Have Reduced Adherence and Cytotoxicity to Human Ectocervical Cells

Identification of Trichomonas vaginalis Cysteine Proteases That Induce Apoptosis in Human Vaginal Epithelial Cells

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