Morphological studies of the cytotoxicity of Trichomonas vaginalis to normal human vaginal epithelial cells in vitro.

ABSTRACT In this study we established human vaginal epithelial cells (hVECs) in culture and evaluated their interaction with Trichomonas vaginalis parasites to complement previous studies using other cell types. Primary cultures of hVECs were established. Contaminating fibroblasts were separated from epithelial cells by differential trypsinization. Specific antibody staining revealed that over 92% of cells in hVEC monolayers were epithelial cells. T. vaginalis adhered to hVECs and produced severe cytotoxic effects resulting in obliteration of the monolayer within 24 h. Adherence and cytotoxicity were not observed when T. vaginalis was exposed to human vaginal fibroblasts or bovine vaginal epithelial cells. Likewise, the bovine parasite Tritrichomonas foetushad no cytotoxic effects on hVECs. We concluded that the interaction between T. vaginalis and hVECs is both cell specific (limited to epithelial cells and not vaginal fibroblasts) and species specific (limited to human vaginal cells and not bovine cells). Pretreatment of T. vaginalis with metronidazole or periodate abolished the adhesion of parasites to cell monolayers and the cytotoxic effect, suggesting involvement of carbohydrate-containing molecules in these processes. Different clinical isolates of T. vaginalis caused damage to cultured cells at different rates. Parasites separated from the vaginal cell monolayer by a permeable membrane did not produce a cytopathic effect, suggesting contact-dependent cytotoxicity.

Download Full-text

Adhesion of Tritrichomonas foetus to Bovine Vaginal Epithelial Cells

Infection and Immunity ◽

10.1128/iai.67.8.3847-3854.1999 ◽

1999 ◽

Vol 67 (8) ◽

pp. 3847-3854 ◽

Cited By ~ 53

Author(s):

B. N. Singh ◽

J. J. Lucas ◽

D. H. Beach ◽

S. T. Shin ◽

R. O. Gilbert

Keyword(s):

Epithelial Cells ◽

Cell Surface ◽

Trichomonas Vaginalis ◽

Host Cells ◽

Tritrichomonas Foetus ◽

Trypan Blue Exclusion ◽

Vaginal Epithelial Cells ◽

Host Parasite

ABSTRACT An in vitro culture system of bovine vaginal epithelial cells (BVECs) was developed to study the cytopathogenic effects ofTritrichomonas foetus and the role of lipophosphoglycan (LPG)-like cell surface glycoconjugates in adhesion of parasites to host cells. Exposure of BVEC monolayers to T. foetusresulted in extensive damage of monolayers. Host cell disruption was measured quantitatively by a trypan blue exclusion assay and by release of 3H from [3H]thymidine-labeled host cells. Results indicated contact-dependent cytotoxicity of host cells byT. foetus. The cytopathogenic effect was a function ofT. foetus density. Metronidazole- or periodate-treatedT. foetus showed no damage to BVEC monolayers. A related human trichomonad, Trichomonas vaginalis, showed no cytotoxic effects, indicating species-specific host-parasite interactions. A direct binding assay was developed and used to investigate the role of a major cell surface LPG-like molecule in host-parasite adhesion. The results of competition experiments showed that the binding to BVECs was displaceable, was saturable, and yielded a typical binding curve, suggesting that specific receptor-ligand interactions mediate the attachment of T. foetus to BVECs. Progesterone-treated BVECs showed enhanced parasite binding. T. foetus LPG inhibited the binding of T. foetus to BVECs; the LPG from T. vaginalis and a variety of other glycoconjugates did not. These data imply specificity of LPG on host-parasite adhesion. Periodate-treated parasites showed no adherence to host cells, indicating the involvement of carbohydrate containing molecules in the adhesion process.

Download Full-text

CP30, a Cysteine Proteinase Involved inTrichomonas vaginalis Cytoadherence

Infection and Immunity ◽

10.1128/iai.68.9.4907-4912.2000 ◽

2000 ◽

Vol 68 (9) ◽

pp. 4907-4912 ◽

Cited By ~ 82

Author(s):

M. Remedios Mendoza-López ◽

Cecilia Becerril-Garcia ◽

Loriz V. Fattel-Facenda ◽

Leticia Avila-Gonzalez ◽

Martha E. Ruíz-Tachiquín ◽

...

Keyword(s):

Epithelial Cells ◽

Hela Cell ◽

Trichomonas Vaginalis ◽

Cysteine Proteinase ◽

Collagen Iv ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cell Monolayers ◽

Thiol Proteinase

ABSTRACT We describe here the participation of a Trichomonas vaginalis 30-kDa proteinase (CP30) with affinity to the HeLa cell surface in attachment of this parasite to host epithelial cells. The CP30 band is a cysteine proteinase because its activity was inhibited by E-64, a thiol proteinase inhibitor. In two-dimensional substrate gel electrophoresis of total extracts of the trichomonad isolate CNCD 147, three spots with proteolytic activity were detected in the 30-kDa region, in the pI range from 4.5 to 5.5. Two of the spots (pI 4.5 and 5.0) bound to the surfaces of fixed HeLa cells corresponding to the CP30 band. The immunoglobulin G fraction of the rabbit anti-CP30 antiserum that recognized a 30-kDa band by Western blotting and immunoprecipitated CP30 specifically inhibited trichomonal cytoadherence to HeLa cell monolayers in a concentration-dependent manner and reacted with CP30 at the parasite surface. CP30 degraded proteins found on the female urogenital tract, including fibronectin, collagen IV, and hemoglobin. Interestingly, CP30 digested fibronectin and collagen IV only at pH levels between 4.5 and 5.0. Moreover, trichomonosis patients whose diagnosis was confirmed by in vitro culture possessed antibody to CP30 in both sera and vaginal washes, and CP30 activity was found in vaginal washes. Our results suggest that surface CP30 is a cysteine proteinase necessary for trichomonal adherence to human epithelial cells.

Download Full-text

Development of highly sensitive and specific in vitro renal solute carrier (SLC) uptake cell models using normal human adult renal proximal tubule epithelial cells for drug transporter interaction studies

Toxicology Letters ◽

10.1016/j.toxlet.2018.06.650 ◽

2018 ◽

Vol 295 ◽

pp. S114-S115

Author(s):

R. Menth ◽

C. Zou ◽

L. Romero ◽

E. Turner ◽

K. Huang ◽

...

Keyword(s):

Epithelial Cells ◽

Proximal Tubule ◽

Renal Proximal Tubule ◽

Drug Transporter ◽

Cell Models ◽

Human Adult ◽

Highly Sensitive ◽

Normal Human ◽

Solute Carrier

Download Full-text

Cigarette smoke condensate-induced transformation of normal human breast epithelial cells in vitro

Oncogene ◽

10.1038/sj.onc.1207792 ◽

2004 ◽

Vol 23 (35) ◽

pp. 5880-5889 ◽

Cited By ~ 49

Author(s):

Satya Narayan ◽

Aruna S Jaiswal ◽

Diana Kang ◽

Pratima Srivastava ◽

Gokul M Das ◽

...

Keyword(s):

Epithelial Cells ◽

Cigarette Smoke ◽

Cigarette Smoke Condensate ◽

Human Breast ◽

Breast Epithelial Cells ◽

Normal Human Breast ◽

Normal Human ◽

Breast Epithelial ◽

Human Breast Epithelial Cells

Download Full-text

Influence of probiotic vaginal lactobacilli on in vitro adhesion of urogenital pathogens to vaginal epithelial cells

Letters in Applied Microbiology ◽

10.1111/j.1472-765x.2006.01934.x ◽

2006 ◽

Vol 43 (2) ◽

pp. 174-180 ◽

Cited By ~ 81

Author(s):

G. Zarate ◽

M.E. Nader-Macias

Keyword(s):

Epithelial Cells ◽

Vaginal Epithelial Cells ◽

Vaginal Lactobacilli ◽

In Vitro Adhesion

Download Full-text

Trichomonas vaginalis adherence mediates differential gene expression in human vaginal epithelial cells

Cellular Microbiology ◽

10.1111/j.1462-5822.2005.00522.x ◽

2005 ◽

Vol 7 (6) ◽

pp. 887-897 ◽

Cited By ~ 24

Author(s):

Ashwini Kucknoor ◽

Vasanthakrishna Mundodi ◽

John F. Alderete

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Differential Gene Expression ◽

Trichomonas Vaginalis ◽

Vaginal Epithelial Cells ◽

Differential Gene

Download Full-text

Trichomonas vaginalis Lipophosphoglycan Triggers a Selective Upregulation of Cytokines by Human Female Reproductive Tract Epithelial Cells

Infection and Immunity ◽

10.1128/iai.00631-06 ◽

2006 ◽

Vol 74 (10) ◽

pp. 5773-5779 ◽

Cited By ~ 100

Author(s):

Raina N. Fichorova ◽

Radiana T. Trifonova ◽

Robert O. Gilbert ◽

Catherine E. Costello ◽

Gary R. Hayes ◽

...

Keyword(s):

Epithelial Cells ◽

Trichomonas Vaginalis ◽

Reproductive Tract ◽

Adaptor Protein ◽

Dendritic Cell Maturation ◽

Female Reproductive Tract ◽

Dependent Manner ◽

Sexually Transmitted ◽

Vaginal Epithelial Cells ◽

Dose Dependent

ABSTRACT Trichomonas vaginalis is one of the most common nonviral sexually transmitted human infections and, worldwide, has been linked to increased incidence of human immunodeficiency virus type 1 transmission, preterm delivery, low birth weight, cervical cancer, and vaginitis. The molecular pathways that are important in initiating host inflammatory and immune responses to T. vaginalis are poorly understood. Here we report interactions of human cervicovaginal epithelial cells with the most abundant cell surface glycoconjugate of the parasite, the T. vaginalis lipophosphoglycan (LPG). Purified LPG mediated the adhesion of parasites to human vaginal epithelial cells in a dose-dependent manner. Furthermore, T. vaginalis LPG (but not LPG from Tritrichomonas foetus, the causative agent of bovine trichomoniasis) induced a selective upregulation of chemotactic cytokines by human endocervical, ectocervical, and vaginal epithelial cells, which do not express Toll-like receptor 4/MD2. The T. vaginalis LPG triggered interleukin 8 (IL-8), which promotes the adhesion and transmigration of neutrophils across the endothelium, and macrophage inflammatory protein 3α, which is a chemoattractant for immune cells and is essential for dendritic cell maturation. These effects were dose dependent and sustained in the absence of cytotoxicity and IL-1β release and utilized, at least in part, a signaling pathway independent from the Toll-like/IL-1 receptor adaptor protein MyD88.

Download Full-text