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

2006 ◽  
Vol 74 (10) ◽  
pp. 5773-5779 ◽  
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.

scholarly journals Immunogenic and Plasminogen-Binding Surface-Associated α-Enolase of Trichomonas vaginalis

2007 ◽  
Vol 76 (2) ◽  
pp. 523-531 ◽  
Author(s):  
V. Mundodi ◽  
A. S. Kucknoor ◽  
J. F. Alderete
Keyword(s):  
Trichomonas Vaginalis ◽  
Specific Inhibitor ◽  
Aminocaproic Acid ◽  
Glycolytic Enzyme ◽  
Cdna Expression Library ◽  
Dependent Manner ◽  
Expression Library ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
Vaginal Epithelial Cells

ABSTRACT Trichomonas vaginalis is a protist that causes the most common human sexually transmitted infection. A T. vaginalis cDNA expression library was screened with pooled sera from patients with trichomoniasis. A highly reactive cDNA clone of 1,428 bp encoded a trichomonad protein of 472 amino acids with sequence identity to α-enolase (tv-eno1). The sequence alignment confirmed the highly conserved nature of the enzyme with 65% to 84% identity among organisms. The expression of tv-eno1 was up-regulated by contact of parasites with vaginal epithelial cells, and this is the first report demonstrating up-regulation by cytoadherence of a plasminogen-binding α-enolase in T. vaginalis. Immunofluorescence with monoclonal antibody of nonpermeabilized trichomonads showed tv-ENO1 on the surface. The recombinant tv-ENO1 was expressed in Escherichia coli as a glutathione S-transferase (GST)::tv-ENO1 fusion protein, which was cleaved using thrombin to obtain affinity-purified recombinant tv-ENO1 protein (tv-rENO1) detectable in immunoblots by sera of patients. Immobilized tv-rENO1 bound human plasminogen in a dose-dependent manner, and plasminogen binding by tv-rENO1 was confirmed in a ligand blot assay. The plasminogen-specific inhibitor ε-aminocaproic acid blocked the tv-rENO1-plasminogen association, indicating that lysines play a role in binding to tv-rENO1. Further, both parasites and tv-rENO1 activate plasminogen to plasmin that is mediated by tissue plasminogen activator. These data indicate that as with other bacterial pathogens, tv-ENO1 is an anchorless, surface-associated glycolytic enzyme of T. vaginalis.

scholarly journals Morphological characterization of vaginal epithelial cells of santa inês ewes subjected to estrus synchronization

2019 ◽  
Vol 10 (1) ◽  
pp. 5-9
Author(s):  
Camila Vasconcelos Ribeiro ◽  
Tábatta Arrivabene Neves ◽  
Glaucia Brandão Fagundes ◽  
Dayana Maria Do Nascimento ◽  
Cleidson Manoel Gomes Da Silva ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Reproductive Tract ◽  
Cell Types ◽  
Morphological Characterization ◽  
Female Reproductive Tract ◽  
Estrus Synchronization ◽  
Essential Information ◽  
Vaginal Epithelial Cells ◽  
Staining Methods ◽  
Intermediate Cells

Vaginal cytology analysis has been used to evaluate the different stages of estrous cycle of several species; it presents a direct correlation with the animal’s hormonal state and provides essential information about the female reproductive tract conditions. Two staining methods were tested to evaluate the vaginal epithelial cell morphology of nulliparous and multiparous ewes during the estrus period. An intravaginal device impregnated with medroxyprogesterone acetate was kept into 10 nulliparous and 10 multiparous ewes for 14 days for estrus synchronization. Then, the progesterone device was withdrawn, and 300 IU of eCG was administered intramuscularly. Vaginal smears were prepared for posterior staining with Panotico or Giemsa stains when estrus was detected. The cells were classified into nucleated superficial, anucleate superficial, intermediate, parabasal, and basal. The Panotico and Giemsa staining of the different cell types studied were satisfactory. A predominance of intermediate epithelial cells (p<0.05) was found after staining. No difference in percentages of the different types of vaginal epithelial cells between nulliparous and multiparous ewes were found. Therefore, both staining methods were efficient, and a predominance of intermediate cells is found in nulliparous and multiparous ewes during the estrus period.

176. GDF-9, BMP-15 AND ACTIVIN A CONTRIBUTE TO SEMINAL FLUID SIGNALLING IN HUMAN CERVICAL EPITHELIAL CELLS

2009 ◽  
Vol 21 (9) ◽  
pp. 94
Author(s):  
D. J. Sharkey ◽  
R. B. Gilchrist ◽  
S. A. Robertson
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Cytokines ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Activin A ◽  
Female Reproductive Tract ◽  
Dependent Manner ◽  
Species Introduction ◽  
Gm Csf ◽  
Pro Inflammatory Cytokines

We have previously shown that in women, as in animal species, introduction of seminal fluid at coitus induces cytokine expression and a local inflammatory-like response in the female reproductive tract. The response is characterised by induction of mRNAs encoding several pro-inflammatory cytokines including GM-CSF, IL-1α and IL-6, as well as chemokines IL-8, MIP-3α and MCP-1. Recent studies in our laboratory have focused on identifying active signalling agents present in human seminal plasma. To date we have shown that all three mammalian isoforms of TGFβ (TGFβ1, TGFβ2 and TGFβ3) and 19-hydroxy PGE are key regulators of this response. The current study aimed to determine whether other members of the TGFβ superfamily including GDF-9, BMP-15 and Activin A also contribute. To investigate this we utilised immortalised Ect1 ectocervical epithelial cells, which mimic the response of primary ectocervical epithelial cells. Ect1 cells were incubated with increasing doses (0.5, 5.0, 50 or 500 ng/ml) of rGDF-9, rBMP-15 or rActivin A and production of pro-inflammatory cytokines and chemokines was assessed in 24 hour post-treatment supernatants using Luminex microbead technology. BMP-15 was found to stimulate GM-CSF production (2-fold), while GDF-9 and Activin A both stimulated IL-6 production (2.4-fold and 80% increases respectively), all acting in a dose-dependent manner. In contrast, all three factors inhibited IL-8 production by Ect1 cells. These data demonstrate that GDF-9, BMP-15 and Activin A are new seminal fluid signalling agents capable of targeting female reproductive tract epithelial cells and inducing different response profiles compared with TGFβ and 19-hydroxy PGE. The relative bioavailability of these factors in seminal fluid would therefore influence the profile of inflammatory response in the female partner, regulating immune responses to male seminal antigens as well as sexually transmitted pathogens.

scholarly journals CP30, a Cysteine Proteinase Involved inTrichomonas vaginalis Cytoadherence

2000 ◽  
Vol 68 (9) ◽  
pp. 4907-4912 ◽  
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.

scholarly journals Cytotoxic Effects of Air Freshener Biocides in Lung Epithelial Cells

2013 ◽  
Vol 8 (9) ◽  
pp. 1934578X1300800
Author(s):  
Jung-Taek Kwon ◽  
Mimi Lee ◽  
Gun-Baek Seo ◽  
Hyun-Mi Kim ◽  
Ilseob Shim ◽  
...  
Keyword(s):  
Dna Damage ◽  
Epithelial Cells ◽  
Cell Viability ◽  
Lung Epithelial Cells ◽  
Ros Generation ◽  
Dependent Manner ◽  
Cytotoxic Effects ◽  
Lung Epithelial ◽  
Dose Dependent ◽  
A549 Lung

This study evaluated the cytotoxicity of mixtures of citral (CTR) and either benzisothiazolinone (BIT, Mix-CTR-BIT) or triclosan (TCS, Mix-CTR-TCS) in human A549 lung epithelial cells. We investigated the effects of various mix ratios of these common air freshener ingredients on cell viability, cell proliferation, reactive oxygen species (ROS) generation, and DNA damage. Mix-CTR-BIT and Mix-CTR-TCS significantly decreased the viability of lung epithelial cells and inhibited cell growth in a dose-dependent manner. In addition, both mixtures increased ROS generation, compared to that observed in control cells. In particular, cell viability, growth, and morphology were affected upon increase in the proportion of BIT or TCS in the mixture. However, comet analysis showed that treatment of cells with Mix-CTR-BIT or Mix-CTR-TCS did not increase DNA damage. Taken together, these data suggested that increasing the content of biocides in air fresheners might induce cytotoxicity, and that screening these compounds using lung epithelial cells may contribute to hazard assessment.

scholarly journals Glyceraldehyde-3-Phosphate Dehydrogenase Is a Surface-Associated, Fibronectin-Binding Protein of Trichomonas vaginalis

2009 ◽  
Vol 77 (7) ◽  
pp. 2703-2711 ◽  
Author(s):  
A. Lama ◽  
A. Kucknoor ◽  
V. Mundodi ◽  
J. F. Alderete
Keyword(s):  
Trichomonas Vaginalis ◽  
Binding Protein ◽  
Transmitted Disease ◽  
Surface Expression ◽  
Cdna Expression Library ◽  
Expression Library ◽  
Sexually Transmitted ◽  
Vaginal Epithelial Cells ◽  
Extracellular Matrix Components ◽  
Fibronectin Binding Protein

ABSTRACT Trichomonas vaginalis colonizes the urogenital tract of humans and causes trichomonosis, the most prevalent nonviral sexually transmitted disease. We have shown an association of T. vaginalis with basement membrane extracellular matrix components, a property which we hypothesize is important for colonization and persistence. In this study, we identify a fibronectin (FN)-binding protein of T. vaginalis. A monoclonal antibody (MAb) from a library of hybridomas that inhibited the binding of T. vaginalis organisms to immobilized FN was identified. The MAb (called ws1) recognized a 39-kDa protein and was used to screen a cDNA expression library of T. vaginalis. A 1,086-bp reactive cDNA clone that encoded a protein of 362 amino acids with identity to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was obtained. The gapdh gene was cloned, and recombinant GAPDH (rGAPDH) was expressed in Escherichia coli cells. Natural GAPDH and rGAPDH bound to immobilized FN and to plasminogen and collagen but not to laminin. MAb ws1 inhibited binding to FN. GAPDH was detected on the surface of trichomonads and was upregulated in synthesis and surface expression by iron. Higher levels of binding to FN were seen for organisms grown in iron-replete medium than for organisms grown in iron-depleted medium. In addition, decreased synthesis of GAPDH by antisense transfection of T. vaginalis gave lower levels of organisms bound to FN and had no adverse effect on growth kinetics. Finally, GAPDH did not associate with immortalized vaginal epithelial cells (VECs), and neither GAPDH nor MAb ws1 inhibited the adherence of trichomonads to VECs. These results indicate that GAPDH is a surface-associated protein of T. vaginalis with alternative functions.

scholarly journals Hyperinsulinemia, But Not Other Factors Associated with Insulin Resistance, Acutely Enhances Colorectal Epithelial Proliferation in Vivo

Endocrinology ◽  
2006 ◽  
Vol 147 (4) ◽  
pp. 1830-1837 ◽  
Author(s):  
Thien T. Tran ◽  
Dinaz Naigamwalla ◽  
Andrei I. Oprescu ◽  
Loretta Lam ◽  
Gail McKeown-Eyssen ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Epithelial Cells ◽  
Dependent Manner ◽  
Epithelial Proliferation ◽  
Euglycemic Clamp ◽  
The Individual ◽  
Dose Dependent ◽  
Circulating Levels

The similarity in risk factors for insulin resistance and colorectal cancer (CRC) led to the hypothesis that markers of insulin resistance, such as elevated circulating levels of insulin, glucose, fatty acids, and triglycerides, are energy sources and growth factors in the development of CRC. The objective was thus to examine the individual and combined effects of these circulating factors on colorectal epithelial proliferation in vivo. Rats were fasted overnight, randomized to six groups, infused iv with insulin, glucose, and/or Intralipid for 10 h, and assessed for 5-bromo-2-deoxyuridine labeling of replicating DNA in colorectal epithelial cells. Intravenous infusion of insulin, during a 10-h euglycemic clamp, increased colorectal epithelial proliferation in a dose-dependent manner. The addition of hyperglycemia to hyperinsulinemia did not further increase proliferation. Intralipid infusion alone did not affect proliferation; however, the combination of insulin, glucose, and Intralipid infusion resulted in greater hyperinsulinemia than the infusion of insulin alone and further increased proliferation. Insulin infusion during a 10-h euglycemic clamp decreased total IGF-I levels and did not affect insulin sensitivity. These results provide evidence for an acute role of insulin, at levels observed in insulin resistance, in the proliferation of colorectal epithelial cells in vivo.

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