Note: This page contains sample records for the topic vaginal epithelial cells from Science.gov.
While these samples are representative of the content of Science.gov,
they are not comprehensive nor are they the most current set.
We encourage you to perform a real-time search of Science.gov
to obtain the most current and comprehensive results.
Last update: November 12, 2013.
1

Regulation of natural antimicrobial defences in human vaginal epithelial cells  

Microsoft Academic Search

BackgroundThe involvement of natural antimicrobial peptides in infection-associated preterm birth is unknown. Our aim was to assess regulation by granulocyte-macrophage colony-stimulating factor, monocyte-chemotactic-protein-1 (GM-CSF and MCP-1, interleukin-1 (IL) and lipopolysaccharide (LPS) in cultured human vaginal epithelial cells.MethodsVK2\\/E6E7 cells were treated with IL-1, MCP-1 and GM-CSF (20\\/100\\/200\\/500 pg\\/ml and 1 ng\\/ml) at 2 h, 4 h, 6 h and 24 h

C Foster; E Chin-Smith; R Tribe

2011-01-01

2

Vaginal and Oral Epithelial Cell Anti-Candida Activity  

Microsoft Academic Search

Candida albicans is the causative agent of acute and recurrent vulvovaginal candidiasis (VVC), a common mucosal infection affecting significant numbers of women in their reproductive years. While any murine host protective role for cell-mediated immunity (CMI), humoral immunity, and innate resistance by neutrophils against the vaginal infection appear negligible, significant in vitro growth inhibition of Candida species by vaginal and

Fatema Nomanbhoy; Chad Steele; Junko Yano

2002-01-01

3

HIV gp120 induced gene expression signatures in vaginal epithelial cells.  

PubMed

Gp120 is the envelope protein of HIV which binds to CD4 independent proteins on vaginal epithelial cells. HIV-gp120 has been reported to modulate gene expression in several cell types. How this interaction may alter the physiologic vaginal milieu during the earliest stages of vaginal transmission of HIV, is currently unknown. Vaginal epithelial cells were treated with HIV-gp120, and a global snapshot of changes in gene expression profiles, were unraveled by microarray analysis. The differentially expressed genes were involved in diverse cellular functions. Genes of immunomodulatory processes and induction of proteases were highly enriched. We propose that the induction of inflammation and proteases may act in concert to weaken the vaginal epithelium, making it more permeable to viral entry. Identification of the gene signatures involved in vaginal-HIV dialogue would aid in understanding the environ induced by HIV itself, as the virus invades and gains entry into its host. PMID:23867815

Fanibunda, Sashaina E; Modi, Deepak N; Bandivdekar, Atmaram H

2013-07-15

4

Tritrichomonas foetus Induces Apoptotic Cell Death in Bovine Vaginal Epithelial Cells  

Microsoft Academic Search

Tritrichomonas foetus is a serious veterinary pathogen, causing bovine trichomoniasis, a sexually transmitted disease leading to infertility and abortion. T. foetus infects the mucosal surfaces of the reproductive tract. Infection with T. foetus leads to apoptotic cell death of bovine vaginal epithelial cells (BVECs) in culture. An affinity-purified cysteine protease (CP) fraction yielding on sodium dodecyl sulfate-polyacrylamide gel elec- trophoresis

B. N. Singh; J. J. Lucas; G. R. Hayes; Ish Kumar; D. H. Beach; Marcel Frajblat; R. O. Gilbert; U. Sommer; C. E. Costello

2004-01-01

5

Oral and vaginal epithelial cell anti-Candida activity is acid labile and does not require live epithelial cells  

PubMed Central

Background: Candida albicans is the causative agent of oral and vaginal candidiasis. Innate host defenses against C. albicans are important against each infection. Among these are oral and vaginal epithelial cells that have anti-Candida activity. The mechanism of action includes a requirement for cell contact with no role for soluble factors, and a putative role for carbohydrates based on the sensitivity of the activity to periodic acid. Methods: Periodic acid treatment of epithelial cells as well as the property of partial resistance of antifungal activity to fixation was used to further dissect the mechanism of action. Results: The results herein effectively now challenge a role for carbohydrates alone. Firstly, the putative carbohydrate(s) released into supernatants of periodic acid-treated epithelial cells could not compete with fresh epithelial cells for activity, and equivalent abrogation of activity was observed by periodic acid-treated cells irrespective of the amount of carbohydrate released. Instead, the similar abrogation of activity following treatment with other acids or when cocultured under acidic conditions suggests that the activity is acid-labile. Finally, while activity requires intact epithelial cells, it does not require live cells; activity was minimally affected by fixing epithelial cells prior to coculture where the majority of cells remained impermeable to Trypan blue but were defined as non–viable by positive nuclear staining with propidium iodide. Conclusion: These results suggest that antifungal activity is dependent on contact by intact, but not necessarily live, epithelial cells through an acid-labile mechanism.

Yano, J.; Lilly, E. A.; Steele, C.; Fortenberry, D.; Fidel, P. L.

2005-01-01

6

Adhesion of Tritrichomonas foetus to Bovine Vaginal Epithelial Cells  

Microsoft Academic Search

release of 3H from (3H)thymidine-labeled host cells. Results indicated contact-dependent cytotoxicity of host cells by T. foetus. The cytopathogenic effect was a function of T. foetus density. Metronidazole- or periodate- treated T. 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

B. N. SINGH; J. J. LUCAS; D. H. BEACH; S. T. SHIN; R. O. GILBERT

7

Candida albicans Adhesion to and Invasion and Damage of Vaginal Epithelial Cells: Stage-Specific Inhibition by Clotrimazole and Bifonazole?  

PubMed Central

Clotrimazole and bifonazole are highly effective antifungal agents against mucosal Candida albicans infections. Here we examined the effects of low levels of clotrimazole and bifonazole on the ability of C. albicans to adhere, invade, and damage vaginal epithelial cells. Although adhesion and invasion were not affected, damage was greatly reduced upon azole treatment. This clearly indicates that low levels of azoles influence specific activities of C. albicans during distinct stages of vaginal epithelium infections.

Wachtler, Betty; Wilson, Duncan; Hube, Bernhard

2011-01-01

8

Evidence for Gardnerella vaginalis uptake and internalization by squamous vaginal epithelial cells: implications for the pathogenesis of bacterial vaginosis.  

PubMed

Bacterial vaginosis (BV), a common condition seen in premenopausal women, is associated with preterm labor, pelvic inflammatory disease, and delivery of low birth weight infants. Gardnerella vaginalis is the predominant bacterial species associated with BV, although its exact role in the pathology of BV is unknown. Using immunofluorescence, confocal and transmission electron microscopy, we found that VK2 vaginal epithelial cells take up G. vaginalis after exposure to the bacteria. Confocal microscopy also indicated the presence of internalized G. vaginalis within vaginal epithelial cells obtained from a subject with BV. Using VK2 cells and (35)S labeled bacteria in an invasion assay, we found that a 1 h uptake of G. vaginalis was 21.8-fold higher than heat-killed G. vaginalis, 84-fold compared to Lactobacillus acidophilus and 6.6-fold compared to Lactobacillus crispatus. Internalization was inhibited by pre-exposure of cells to cytochalasin-D. In addition, the cytoskeletal protein vimentin was upregulated in VK2 cells exposed to G. vaginalis, but there was no change in actin cytoskeletal polymerization/rearrangements or vimentin subcellular relocalization post exposure. Cytoskeletal protein modifications could represent a potential mechanism for G. vaginalis mediated internalization by vaginal epithelial cells. Finally, understanding vaginal bacteria/host interactions will allow us to better understand the underlying mechanisms of BV pathogenesis. PMID:22227318

Marrs, Christy N; Knobel, Susan M; Zhu, Wen Qin; Sweet, Stephanie D; Chaudhry, Ahsen R; Alcendor, Donald J

2011-12-24

9

Evidence for Gardnerella vaginalis uptake and internalization by squamous vaginal epithelial cells: implications for the pathogenesis of bacterial vaginosis  

PubMed Central

Bacterial vaginosis (BV), a common condition seen in premenopausal women, is associated with preterm labor, pelvic inflammatory disease, and delivery of low birth weight infants. Gardnerella vaginalis is the predominant bacterial species associated with BV, although its exact role in the pathology of BV is unknown. Using immunofluorescence, confocal and transmission electron microscopy, we found that VK2 vaginal epithelial cells take up G. vaginalis after exposure to the bacteria. Confocal microscopy also indicated the presence of internalized G. vaginalis within vaginal epithelial cells obtained from a subject with BV. Using VK2 cells and 35S labeled bacteria in an invasion assay, we found that a 1 h uptake of G. vaginalis was 21.8-fold higher than heat-killed G. vaginalis, 84-fold compared to Lactobacillus acidophilus and 6.6-fold compared to Lactobacillus crispatus. Internalization was inhibited by pre-exposure of cells to cytochalasin-D. In addition, the cytoskeletal protein vimentin was upregulated in VK2 cells exposed to G. vaginalis, but there was no change in actin cytoskeletal polymerization/rearrangements or vimentin subcellular relocalization post exposure. Cytoskeletal protein modifications could represent a potential mechanism for G. vaginalis mediated internalization by vaginal epithelial cells. Finally, understanding vaginal bacteria/host interactions will allow us to better understand the underlying mechanisms of BV pathogenesis.

Marrs, Christy N.; Knobel, Susan M.; Zhu, Wen Qin; Sweet, Stephanie D.; Chaudhry, Ahsen R.; Alcendor, Donald J.

2012-01-01

10

Enterococcus faecalis inhibits superantigen toxic shock syndrome toxin-1-induced interleukin-8 from human vaginal epithelial cells through tetramic acids.  

PubMed

The vaginal mucosa can be colonized by many bacteria including commensal organisms and potential pathogens, such as Staphylococcus aureus. Some strains of S. aureus produce the superantigen toxic shock syndrome toxin-1, which can penetrate the vaginal epithelium to cause toxic shock syndrome. We have observed that a female was mono-colonized with Enterococcus faecalis vaginally as tested in aerobic culture, even upon repeated culture for six months, suggesting this organism was negatively influencing colonization by other bacteria. In recent studies, we demonstrated an "outside-in" mechanism of cytokine signaling and consequent inflammation that facilitates the ability of potential pathogens to initiate infection from mucosal surfaces. Thus, we hypothesized that this strain of E. faecalis may make anti-inflammatory factors which block disease progression of more pathogenic organisms. E. faecalis MN1 inhibited interleukin-8 production from human vaginal epithelial cells in response to the vaginal pathogens Candida albicans, Gardnerella vaginalis, and Neisseria gonorrhoeae, as well as to toxic shock syndrome toxin-1. We further demonstrated that this organism secretes two tetramic acid compounds which appear responsible for inhibition of interleukin-8 production, as well as inhibition of T cell proliferation due to toxic shock syndrome toxin-1. Microbicides that include anti-inflammatory molecules, such as these tetramic acid compounds naturally produced by E. faecalis MN1, may be useful in prevention of diseases that develop from vaginal infections. PMID:23613823

Brosnahan, Amanda J; Merriman, Joseph A; Salgado-Pabón, Wilmara; Ford, Bradley; Schlievert, Patrick M

2013-04-22

11

Infection of vaginal and colonic epithelial cells by the human immunodeficiency virus type 1 is neutralized by antibodies raised against conserved epitopes in the envelope glycoprotein gp120.  

PubMed Central

The rectal and genital tract mucosae are considered to be major sites of entry for the human immunodeficiency virus (HIV) during sexual contact. We now demonstrate that vaginal epithelial cells can be infected by HIV type 1 (HIV-1) via a mechanism similar to that described for neuroglial cells and, more recently, for colorectal epithelial cells, involving initial interaction of the HIV-1 envelope glycoprotein gp120 with a cell-surface glycosphingolipid (sulfated lactosylceramide). A hyperimmune serum against gp120 was able to neutralize HIV-1 infection of vaginal epithelial cells. Site-directed immunization was employed to identify sites on gp120 recognized by antibodies neutralizing HIV-1 infection of vaginal and colonic epithelial cells. Hyperimmune sera were raised in monkeys against a series of 40 overlapping synthetic peptides covering the entire sequence of HIV-1 (HTLV-IIIB) gp120. Antisera raised against five synthetic peptides, corresponding to three relatively conserved regions and to the hypervariable region (V3 loop), efficiently neutralized HIV-1 infection of human vaginal epithelial cells in vitro. Similar results were obtained with the colonic cells. Hyperimmune sera to all five peptides have been shown earlier to neutralize HIV-1 infectivity in CD4+ T cells. These results have obvious implications for the design of mucosal subunit vaccines against sexually transmitted HIV-1 infections. Images Fig. 1 Fig. 2 Fig. 3

Furuta, Y; Eriksson, K; Svennerholm, B; Fredman, P; Horal, P; Jeansson, S; Vahlne, A; Holmgren, J; Czerkinsky, C

1994-01-01

12

Effect of the Purinergic Receptor P2X7 on Chlamydia Infection in Cervical Epithelial Cells and Vaginally Infected Mice1  

Microsoft Academic Search

Ligation of the purinergic receptor, P2X7R, with its agonist ATP has been previously shown to inhibit intracellular infection by chlamydiae and mycobacteria in macrophages. The effect of P2X7R on chlamydial infection had never been investigated in the preferred target cells of chlamydiae, cervical epithelial cells, nor in vaginally infected mice. In this study, we show that treatment of epithelial cells

Toni Darville; Lynn Welter-Stahl; Cristiane Cruz; Ali Abdul Sater; Charles W. Andrews; David M. Ojcius

2007-01-01

13

Estrogen Acidifies Vaginal pH by Up-Regulation of Proton Secretion via the Apical Membrane of Vaginal-Ectocervical Epithelial Cells  

PubMed Central

The objective of this study was to assess estrogen-dependent cellular mechanisms that could contribute to the acid pH of the vaginal lumen. Cultures of normal human cervical-vaginal epithelial (hECE) cells and endocervical cells were grown on filters, and acidification of the extracellular solutions on the luminal (L-pHo) and contraluminal (CL-pHo) sides was measured. The hECE cells and endocervical cells decreased CL-pHo from 7.40 to 7.25 within 20–30 min of incubation in basic salt solution. Endocervical cells also produced a similar decrease in L-pHo. In contrast, hECE cells acidified L-pHo down to pH 7.05 when grown as monoculture and down to pH 6.05 when grown in coculture with human cervical fibroblasts. This enhanced acid secretion into the luminal compartment was estrogen dependent because removal of endogenous steroid hormones attenuated the effect, whereas treatment with 1 7?-estradiol restored it. The 17?-estradiol effect was dose dependent (EC50 0.5 nM) and could be mimicked by diethyl-stilbestrol and in part by estrone and tamoxifen. Preincubation with ICI-182780, but not with progesterone, blocked the estrogen effect. Preincubation of cells with the V-ATPase blocker bafilomycin A1, when administered to the luminal solution, attenuated the baseline and estrogen-dependent acid secretion into the luminal solution. Treatment with EGTA, to abrogate the tight junctional resistance, blocked the decrease in L-pHo and stimulated a decrease in CL-pHo, indicating that the tight junctions are necessary for maintaining luminal acidification. We conclude that vaginal-ectocervical cells acidify the luminal canal by a mechanism of active proton secretion, driven in part by V-H+-ATPase located in the apical plasma membrane and that the baseline active net proton secretion occurs constitutively throughout life and that this acidification is up-regulated by estrogen.

Gorodeski, George I.; Hopfer, Ulrich; Liu, Chung Chiun; Margles, Ellen

2008-01-01

14

Cytokine and Chemokine Production by Human Oral and Vaginal Epithelial Cells in Response to Candida albicans  

Microsoft Academic Search

Oropharyngeal and vaginal candidiases are the most common forms of mucosal fungal infections and are primarily caused by Candida albicans, a dimorphic fungal commensal organism of the gastrointestinal and lower female reproductive tracts. Clinical and experimental observations suggest that local immunity is important in host defense against candidiasis. Accordingly, cytokines and chemokines are present at the oral and vaginal mucosa

Chad Steele

2002-01-01

15

Epithelial cell-derived S100 calcium-binding proteins as key mediators in the hallmark acute neutrophil response during Candida vaginitis.  

PubMed

Vulvovaginal candidiasis (VVC), caused by Candida species, is a significant problem in women of childbearing age. Similar to clinical observations, a robust vaginal polymorphonuclear neutrophil (PMN) migration occurs in a subset of mice without affecting vaginal fungal burden. We hypothesize that the vaginal PMN infiltrate and accompanying inflammation are not protective but instead are responsible for the symptoms of infection. The purpose of this study was to identify the signal(s) associated with the PMN response in the established mouse model. Vaginal lavage fluid from inoculated mice were categorized base on PMN counts, evaluated for PMN chemotactic activity and analyzed by SDS-PAGE and mass spectrometry (MS) for unique protein identification. The lavage fluid from inoculated mice with high, but not low, PMN levels showed increased chemotactic activity. Likewise, SDS-PAGE of lavage fluid with high PMN levels showed distinct protein patterns. MS revealed that bands at 6 and 14 kDa matched the PMN chemotactic calcium-binding proteins (CBPs), S100A8 and S100A9, respectively. The presence of the CBPs in lavage fluid was confirmed by Western blots and enzyme-linked immunosorbent assay. Vaginal tissues and epithelial cells from inoculated mice with high PMN levels stained more intensely and exhibited increased mRNA transcripts for both proteins compared to those in mice with low PMN levels. Subsequent antibody neutralization showed significant abrogation of the chemotactic activity when the lavage fluid was treated with anti-S100A8, but not anti-S100A9, antibodies. These results reveal that the PMN chemotactic CBP S100A8 and S100A9 are produced by vaginal epithelial cells following interaction with Candida and that S100A8 is a strong candidate responsible for the robust PMN migration during experimental VVC. PMID:20823201

Yano, Junko; Lilly, Elizabeth; Barousse, Melissa; Fidel, Paul L

2010-09-07

16

Development of epithelial and mesenchymal regionalization of the human fetal utero-vaginal anlagen.  

PubMed

Literature on the development of the human vagina is abundant; however, contributions concerning the prenatal development of the entire utero-vaginal anlagen (UVA) are rare or carried out in rodents. The primary epithelial characteristics in the adult vagina and uterus are determined during prenatal development and depend on epithelio-mesenchymal stroma interaction; thus an investigation summarizing the spatiotemporal distribution of relevant molecular markers in the entire human UVA will be of current interest. We phenotyped epithelial and mesenchymal characteristics in sagittal sections from 24 female fetuses of 14-34 weeks of gestation and two female newborns by immunostaining with cytokeratins 8, 13, 14 and 17, p63, bcl-2, bmp4, HOX A13, CD31, VEGF, SMA, Pax2 and vimentin. Epithelial differentiation followed a caudal-to-cranial direction in the UVA. Due to the cytokeratin profile of cytokeratins 8, 13 and 14, the characteristics of the different epithelial zones in the UVA could already be recognized in middle-age fetuses. Vaginal epithelium originated from the urogenital sinus in the lower portion and initiated the transformation of vimentin-positive Müllerian epithelium in the upper vaginal portion. During prenatal development the original squamo-columnar junction was clearly detectable from week 24 onwards and was always found in the cervical canal. Early blc-2 positivity within the surrounding mesenchyme of the entire vagina including the portio region pointed to an organ-specific mesenchymal influence. Prenatal findings in human specimens clearly show that fornix epithelium up to the squamo-columnar junction is of vaginal Müllerian origin, and the cervical epithelium cranial to the squamo-columnar junction is of uterine Müllerian origin and includes cells with enough plasticity to transform into squamous epithelium. PMID:23406280

Fritsch, Helga; Hoermann, Romed; Bitsche, Mario; Pechriggl, Elisabeth; Reich, Olaf

2013-02-13

17

Probiotic lactobacillus and estrogen effects on vaginal epithelial gene expression responses to Candida albicans  

PubMed Central

Background Vaginal epithelial cells have receptors, signal transduction mechanisms, and cytokine secretion capabilities to recruit host defenses against Candida albicans infections. This research evaluates how probiotic lactobacilli affect the defensive epithelial response. Methods This study used quantitative reverse transcription-polymerase chain reaction assay (qRT-PCR), flow cytometry, and a multiplex immunoassay to observe changes in the regulation of gene expression related to cytokine responses in the VK2 (E6/E7) vaginal epithelial cell line treated with 17?-estradiol, exposed to probiotic Lactobacillus rhamnosus GR-1® and Lactobacillus reuteri RC-14® and challenged with C. albicans. Data were statistically evaluated by repeated measures analysis of variance and paired t-tests where appropriate. Results C. albicans induced mRNA expression of genes related to inflammatory cytokine responses associated with nuclear factor-kappa B (NF-?B) and mitogen-activated protein kinase (MAPK) signal transduction pathways. 17?-estradiol suppressed expression of interleukin-1? (IL-1?), IL-6, IL-8, and tumor necrosis factor alpha (TNF?) mRNA. Probiotic lactobacilli suppressed C. albicans-induced nuclear factor-kappa B inhibitor kinase kinase alpha (I???), Toll-like receptor-2 (TLR2), TLR6, IL-8, and TNF?, also suggesting inhibition of NF-?B signaling. The lactobacilli induced expression of IL-1?, and IL-1? mRNA, which was not inhibited by curcumin, suggesting that they induce an alternate inflammatory signal transduction pathway to NF-?B, such as the mitogen activated protein kinase and activator protein-1 (MAPK/AP-1) signal transduction pathway. Curcumin inhibited IL-13 secretion, suggesting that expression of this cytokine is mainly regulated by NF-?B signaling in VK2 cells. Conclusions The results suggest that C. albicans infection induces pro-inflammatory responses in vaginal epithelial cells, and estrogen and lactobacilli suppress expression of NF-?B-related inflammatory genes. Probiotic lactobacilli may induce IL-1? and IL-1? expression by an alternate signal transduction pathway, such as MAPK/AP-1. Activation of alternate signaling mechanisms by lactobacilli to modify epithelial cell cytokine production may be a mechanism for probiotic modulation of morbidity in vulvovaginal candidiasis.

2012-01-01

18

Spindle Cell Epithelioma: A Rare Vaginal Tumor -A Clinico Pathologic Report  

PubMed Central

Spindle cell epithelioma is a very rare benign tumour of the vagina, which contains epithelial and mesenchymal components and co-expresses the markers for both. It has its origin in the epithelial cells of the remnants of the vestibular gland. The presence of glandular structures and the pattern of immunostaining, help in the differentiation of these tumours from the other common vaginal tumours.

K., Nivedita; Sowmya; Shanthini, Fatima

2013-01-01

19

Tubulo-squamous vaginal polyp with basaloid epithelial differentiation.  

PubMed

Tubulo-squamous polyp is a recently described and apparently benign lesion that most frequently involves the upper vagina of postmenopausal patients. Histologically, it is characterized by the presence of cysts, squamous epithelial nests, and small tubular structures that are surrounded by a hypocellular fibrous stroma. Some lesions show focal immunoreactivity for prostatic acid phosphatise and/or prostate-specific antigen raising the possibility of derivation from Skene's glands. In this report, an additional case of tubulo-squamous polyp is presented in which there was prominent basaloid epithelial differentiation, mainly in the form of elongated cords of cells around the peripheral and deep aspect of the lesion. This feature has not been recorded previously in tubulo-squamous polyp and potentially could create diagnostic difficulty with other lesions such as basal cell/adenoid basal carcinoma or small-cell neuroendocrine carcinoma. PMID:19851205

Stewart, Colin J R

2009-11-01

20

Optical Coherence Tomography Compared With Colposcopy for Assessment of Vaginal Epithelial Damage: A Randomized Controlled Trial  

PubMed Central

Objective Colposcopy has been used to detect epithelial damage with vaginal microbicides. In animal models, optical coherence tomography (OCT) provided increased sensitivity over colposcopy in detecting epithelial injury. This randomized double-blinded clinical study compared OCT to colposcopy for the evaluation of epithelial injury in women using placebo or nonoxynol-9. Methods Thirty women aged 18–45 were randomized to use hydroxyethyl cellulose placebo or nonoxynol-9 vaginal gel twice daily for 5.5 days. Imaging with colposcopy and OCT was performed prior to product use, after the last dose, and 1 week later. Colposcopy was graded using standard criteria. OCT images were scored for epithelial integrity based on a published scoring system and measured for epithelial thickness. Results Colposcopy findings and OCT scores and epithelial thicknesses were similar between treatment groups at baseline. After treatment, there were significant differences between the nonoxynol-9 (1.37) and control group (1.15) OCT scores (p<0.001, indicating epithelial injury, and there was epithelial thinning in the nonoxynol-9 group (237?m) compared to the control group (292?m) (p=0.008). There were no significant posttreatment colposcopic differences in epithelial disruption between treatment groups, with only increased erythema noted after nonoxynol-9 use (p=0.02). Conclusion OCT detected epithelial disruption and thinning not identified by colposcopy. Vaginal epithelial thickness, a measure previously available only through biopsy, decreased after nonoxynol-9 use, a finding that may contribute to increased susceptibility to HIV after frequent use. OCT shows promise for the noninvasive clinical assessment of vaginal epithelial damage. Clinical Trial Registration UMIN Clinical Trials Registry, www.umin.ac.jp/ctr/index.htm, R000006186.

Vincent, Kathleen L.; Stanberry, Lawrence R.; Moench, Thomas R.; Breitkopf, Carmen Radecki; Loza, Melissa L.; Wei, Jingna; Grady, James; Paull, Jeremy; Motamedi, Massoud; Rosenthal, Susan L.

2011-01-01

21

Comparative In Vitro Sensitivities of Human Immune Cell Lines, Vaginal and Cervical Epithelial Cell Lines, and Primary Cells to Candidate Microbicides Nonoxynol 9, C31G, and Sodium Dodecyl Sulfate  

Microsoft Academic Search

In experiments to assess the in vitro impact of the candidate microbicides nonoxynol 9 (N-9), C31G, and sodium dodecyl sulfate (SDS) on human immune and epithelial cell viability, cell lines and primary cell populations of lymphocytic and monocytic origin were generally shown to be equally sensitive to exposures ranging from 10 min to 48 h. However, U-937 cells were more

Fred C. Krebs; Shendra R. Miller; Bradley J. Catalone; Raina Fichorova; Deborah Anderson; Daniel Malamud; Mary K. Howett; Brian Wigdahl

2002-01-01

22

Vaginal epithelial surface appearances in women using vaginal rings for contraception  

Microsoft Academic Search

Vaginal inspections using colposcopy before insertion of contraceptive vaginal rings and at 2-month intervals during ring use were conducted on 169 users of four different formulations. The rings studied released Nestorone® alone (50, 75, 100 ?g daily over 6 months); ethinyl estradiol: Nestorone (30:100 and 15:150 ?g daily over 6 months); ethinylestradiol:norethindrone acetate (20:1000 and 15:1000 ?g daily over 4

Ian S Fraser; Maria Lacarra; Daniel R Mishell Jr; Francisco Alvarez; Vivian Brache; Pekka Lähteenmäki; Kaisa Elomaa; Edith Weisberg; Harold A Nash

2000-01-01

23

Surveillance for recurrent cancers and vaginal epithelial lesions in patients with invasive cervical cancer after hysterectomy: are vaginal cytology and high-risk human papillomavirus testing useful?  

PubMed

Objectives: To examine whether women who have had a hysterectomy for cervical cancer may be at an increased risk of vaginal epithelial lesions. Methods: We studied 147 patients with invasive cervical carcinoma (76 squamous cell carcinomas [SCCs], 60 adenocarcinomas [ADCs], and 11 adenosquamous cell carcinomas) who were treated by hysterectomy and had vaginal pathologic follow-up for a mean period of 43.3 months. Results: Of the patients, 15.0% (22/147) developed vaginal intraepithelial neoplasia (VAIN) or recurrence after hysterectomy, including two recurrent carcinomas and eight high-grade VAINs. More important, these high-grade VAINs or recurrent carcinomas were detected only in patients with cervical SCC within the first two years after hysterectomy but not in patients with cervical ADC. Eleven (23.4%) of 47 patients had at least one positive high-risk human papillomavirus (hrHPV) testing result during the follow-up period, and VAIN was detected in 54.5% (6/11) of patients with an hrHPV-positive result compared with 16.7% (6/36) with an hrHPV-negative result. Conclusions: Our results indicate that women with cervical cancer are at an increased risk of VAIN besides recurrence, and women with cervical SCC are more prone to high-grade VAIN/recurrence, especially within the first two years after hysterectomy. The significantly increased detection rate of VAINs/recurrence in the hrHPV-positive group suggests vaginal cytology and HPV cotesting might be the preferred method for surveillance in these women. PMID:24124151

Li, Zaibo; Barron, Stacey; Hong, Wei; Karunamurthy, Arivarasan; Zhao, Chengquan

2013-11-01

24

Gastric Epithelial Stem Cells  

PubMed Central

Advances in our understanding of stem cells in the gastrointestinal tract include the identification of molecular markers of stem and early progenitor cells in the small intestine. Although gastric epithelial stem cells have been localized, little is known about their molecular biology. Recent reports describe the use of inducible Cre recombinase activity to indelibly label candidate stem cells and their progeny in the distal stomach, (ie, the antrum and pylorus). No such lineage labeling of epithelial stem cells has been reported in the gastric body (corpus). Among stem cells in the alimentary canal, those of the adult corpus are unique in that they lie close to the lumen and increase proliferation following loss of a single mature progeny lineage, the acid-secreting parietal cell. They are also unique in that they neither depend on Wnt signaling nor express the surface marker Lgr5. Because pathogenesis of gastric adenocarcinoma has been associated with abnormal patterns of gastric differentiation and with chronic tissue injury, there has been much research on the response of stomach epithelial stem cells to inflammation. Chronic inflammation, as induced by infection with Helicobacter pylori, affects differentiation and promotes metaplasias. Several studies have identified cellular and molecular mechanisms in spasmolytic polypeptide–expressing (pseudopyloric) metaplasia. Researchers have also begun to identify signaling pathways and events that take place during embryonic development that eventually establish the adult stem cells to maintain the specific features and functions of the stomach mucosa. We review the cytologic, molecular, functional, and developmental properties of gastric epithelial stem cells.

MILLS, JASON C.; SHIVDASANI, RAMESH A.

2013-01-01

25

Epithelial Cell Adhesion Molecule  

PubMed Central

The epithelial cell adhesion molecule (EpCAM, CD326) is a glycoprotein of ?40 kd that was originally identified as a marker for carcinoma, attributable to its high expression on rapidly proliferating tumors of epithelial origin. Normal epithelia express EpCAM at a variable but generally lower level than carcinoma cells. In early studies, EpCAM was proposed to be a cell-cell adhesion molecule. However, recent insights revealed a more versatile role for EpCAM that is not limited only to cell adhesion but includes diverse processes such as signaling, cell migration, proliferation, and differentiation. Cell surface expression of EpCAM may actually prevent cell-cell adhesion. Here, we provide a comprehensive review of the current knowledge on EpCAM biology in relation to other cell adhesion molecules. We discuss the implications of the newly identified functions of EpCAM in view of its prognostic relevance in carcinoma, inflammatory pathophysiology, and tissue development and regeneration as well as its role in normal epithelial homeostasis.

Trzpis, Monika; McLaughlin, Pamela M.J.; de Leij, Lou M.F.H.; Harmsen, Martin C.

2007-01-01

26

Vaginitis  

MedlinePLUS

... diagnosed? • How is vaginitis treated? • What is a yeast infection? • What causes yeast infections? • What factors increase the risk of getting a yeast infection? • What are the symptoms of a yeast ...

27

Gastric epithelial stem cells.  

PubMed

Advances in our understanding of stem cells in the gastrointestinal tract include the identification of molecular markers of stem and early progenitor cells in the small intestine. Although gastric epithelial stem cells have been localized, little is known about their molecular biology. Recent reports describe the use of inducible Cre recombinase activity to indelibly label candidate stem cells and their progeny in the distal stomach, (ie, the antrum and pylorus). No such lineage labeling of epithelial stem cells has been reported in the gastric body (corpus). Among stem cells in the alimentary canal, those of the adult corpus are unique in that they lie close to the lumen and increase proliferation following loss of a single mature progeny lineage, the acid-secreting parietal cell. They are also unique in that they neither depend on Wnt signaling nor express the surface marker Lgr5. Because pathogenesis of gastric adenocarcinoma has been associated with abnormal patterns of gastric differentiation and with chronic tissue injury, there has been much research on the response of stomach epithelial stem cells to inflammation. Chronic inflammation, as induced by infection with Helicobacter pylori, affects differentiation and promotes metaplasias. Several studies have identified cellular and molecular mechanisms in spasmolytic polypeptide-expressing (pseudopyloric) metaplasia. Researchers have also begun to identify signaling pathways and events that take place during embryonic development that eventually establish the adult stem cells to maintain the specific features and functions of the stomach mucosa. We review the cytologic, molecular, functional, and developmental properties of gastric epithelial stem cells. PMID:21144849

Mills, Jason C; Shivdasani, Ramesh A

2010-12-07

28

Vaginal T lymphocyte population kinetics during experimental vaginal candidosis: evidence for a possible role of CD8+ T cells in protection against vaginal candidosis  

PubMed Central

Vaginal candidosis represents a significant health problem to women of childbearing age worldwide. It has been postulated that localized T cells play a role in protection against vaginal candidosis. In an attempt to evaluate the role of vaginal T cells in protection against vaginal candidosis, T cell population kinetics was evaluated using an oestrogen-dependent vaginal candidosis murine model. Vaginal T lymphocytes were isolated at different time points post C. albicans inoculation, viable cells were enumerated, phenotypically analysed for the expression of CD3, CD4 and CD8 T cell markers and absolute numbers of T cell subsets were calculated. Oestrogen-induced persistence of vaginal candidosis resulted in a significant increase in the total number of vaginal lymphocytes within 24–48 h post infection; increased vaginal lymphocyte numbers persisted throughout the infection period. The number of CD3+ T cells dramatically increased following C. albicans administration and was maintained at high levels throughout the infection period. The majority of CD3+ T cells were of the CD8+ type; however, considerable numbers of both CD4+ T cells and CD4+CD8+ T cells were also observed throughout the infection period. The considerable and persistent increase in vaginal T cell numbers in general and that of CD8+ T cells in particular are evidence of the possible role played by localized T cells in protection against vaginal candidosis.

GHALEB, M; HAMAD, M; ABU-ELTEEN, K H

2003-01-01

29

Diethylstilbestrol induces vaginal adenosis by disrupting SMAD/RUNX1-mediated cell fate decision in the Müllerian duct epithelium.  

PubMed

Women exposed to diethylstilbestrol (DES) in utero frequently develop vaginal adenosis, from which clear cell adenocarcinoma can arise. Despite decades of extensive investigation, the molecular pathogenesis of DES-associated vaginal adenosis remains elusive. Here we report that DES induces vaginal adenosis by inhibiting the BMP4/Activin A-regulated vaginal cell fate decision through a downregulation of RUNX1. BMP4 and Activin A produced by vaginal mesenchyme synergistically activated the expression of ?Np63, thus deciding vaginal epithelial cell fate in the Müllerian duct epithelial cells (MDECs) via direct binding of SMADs on the highly conserved 5' sequence of ?Np63. Therefore, mice in which Smad4 was deleted in MDECs failed to express ?Np63 in vaginal epithelium and developed adenosis. This SMAD-dependent ?Np63 activation required RUNX1, a binding partner of SMADs. Conditional deletion of Runx1 in the MDECs induced adenosis in the cranial portion of vagina, which mimicked the effect of developmental DES-exposure. Furthermore, neonatal DES exposure downregulated RUNX1 in the fornix of the vagina, where DES-associated adenosis is frequently found. This observation strongly suggests that the downregulation of RUNX1 is the cause of vaginal adenosis. However, once cell fate was determined, the BMP/Activin-SMAD/RUNX1 signaling pathway became dispensable for the maintenance of ?Np63 expression in vaginal epithelium. Instead, the activity of the ?Np63 locus in vaginal epithelium was maintained by a ?Np63-dependent mechanism. This is the first demonstration of a molecular mechanism through which developmental chemical exposure causes precancerous lesions by altering cell fate. PMID:23830984

Laronda, Monica M; Unno, Kenji; Ishi, Kazutomo; Serna, Vanida A; Butler, Lindsey M; Mills, Alea A; Orvis, Grant D; Behringer, Richard R; Deng, Chuxia; Sinha, Satrajit; Kurita, Takeshi

2013-07-04

30

Integrin signaling in epithelial cells  

Microsoft Academic Search

Although most cells of adult mammals express multiple different integrins, particular types of cells have a characteristic repertoire of integrin expression. Benign and malignant epithelial cells use specific integrins to allow the epithelial microenvironment to modulate a wide variety of cell functions, including cell survival, proliferation, morphogenesis, differentiation, motility, invasion and metastasis. An important concept emerging from the data on

Michael Z. Gilcrease

2007-01-01

31

Gynecologic bleeding revealing vaginal metastasis of renal cell carcinoma  

PubMed Central

Vaginal metastases of renal cell carcinoma have been rarely described. We report a case of a 75-year old woman, who underwent radical right nephrectomy for a renal cell carcinoma. Tumour was classified pT3bN0M0 and grade III of Furhmann grading. One year later, scanner discovered mediastinal and lombo-aortic lymph nodes. She received 2 months of immunotherapy associated with bevacizumab, but stopped because of intolerance. She was readmitted in our institute for vaginal bleeding. Clinical investigations showed a vaginal mass and biopsy revealed a renal cell carcinoma metastasis. This case suggests that retrograde venous dissemination may be at the origin of vaginal metastasis of renal cell carcinoma and emphasized the preventive value of early ligature of renal vein.

Benbrahim, Zineb; Chouaib, Ali; Mazeron, Renaud; Leger-Ravet, Marie Benedicte; Lefort, Catherine; Lhomme, Catherine; El Mesbahi, Omar; Escudier, Bernard

2013-01-01

32

Pathogenesis of urinary tract infection —experimental studies of vaginal resistance to colonization  

Microsoft Academic Search

The review summarizes studies of vaginal colonization resistance againstEscherichia coli in a primate model. The genital flora surrounding the urethral orifice exerts a strong colonization resistance. Amoxicillin profoundly disturbs the normal vaginal microflora, reduces its adherence to vaginal epithelial cells in vivo and promotes a persistent vaginalE. coli colonization. Certain cephalosporins may have a similar effect. The induced ecological changes

Jan Winberg; Maria Herthelius-Elman; Roland Miillby; Carl Erik Nord

1993-01-01

33

Response of corneal epithelial cells to Staphylococcus aureus  

PubMed Central

Staphylococcus aureus is a leading cause of invasive infection. It also infects wet mucosal tissues including the cornea and conjunctiva. Conflicting evidence exists on the expression of Toll-like receptors by human corneal epithelial cells. It was therefore of interest to determine how epithelial cells from this immune privileged tissue respond to S. aureus. Further, it was of interest to determine whether cytolytic toxins, with the potential to cause ion flux or potentially permit effector molecule movement across the target cell membrane, alter the response. Microarrays were used to globally assess the response of human corneal epithelial cells to S. aureus. A large increase in abundance of transcripts encoding the antimicrobial dendritic cell chemokine, CCL20, was observed. CCL20 release into the medium was detected, and this response was found to be largely TLR2 and NOD2 independent. Corneal epithelial cells also respond to S. aureus by increasing the intracellular abundance of mRNA for inflammatory mediators, transcription factors, and genes related to MAP kinase pathways, in ways similar to other cell types. The corneal epithelial cell response was surprisingly unaffected by toxin exposure. Toxin exposure did, however, induce a stress response. Although model toxigenic and non-toxigenic strains of S. aureus were employed in the present study, the results obtained were strikingly similar to those reported for stimulation of vaginal epithelial cells by clinical toxic shock toxin expressing isolates, demonstrating that the initial epithelial cellular responses to S. aureus are largely independent of strain as well as epithelial cell tissue source.

2010-01-01

34

Vaginal Microbiome and Epithelial Gene Array in PostMenopausal Women with Moderate to Severe Dryness  

Microsoft Academic Search

After menopause, many women experience vaginal dryness and atrophy of tissue, often attributed to the loss of estrogen. An understudied aspect of vaginal health in women who experience dryness due to atrophy is the role of the resident microbes. It is known that the microbiota has an important role in healthy vaginal homeostasis, including maintaining the pH balance and excluding

Ruben Hummelen; Jean M. Macklaim; Jordan E. Bisanz; Jo-Anne Hammond; Amy McMillan; Rebecca Vongsa; David Koenig; Gregory B. Gloor; Gregor Reid

2011-01-01

35

Ion Channels in Epithelial Cells  

NASA Astrophysics Data System (ADS)

Ion channels in epithelial cells serve to move ions, and in some cases fluid, between compartments of the body. This function of the transfer of material is fundamentally different from that of the transfer of information, which is the main job of most channels in excitable cells. Nevertheless the basic construction of the channels is similar in many respects in the two tissue types. This chapter reviews the nature of channels in epithelia and discusses how their functions have evolved to accomplish the basic tasks for which they are responsible. I will focus on three channel types: epithelial Na+ channels, inward-rectifier K+ channels, and CFTR Cl- channels.

Palmer, Lawrence G.

36

Decreased cervical epithelial sensitivity to nonoxynol-9 (N-9) after four daily applications in a murine model of topical vaginal microbicide safety  

PubMed Central

Background The disappointing clinical failures of five topical vaginal microbicides have provided new insights into factors that impact microbicide safety and efficacy. Specifically, the greater risk for human immunodeficiency virus type 1 (HIV-1) acquisition associated with multiple uses of a nonoxynol-9 (N-9)-containing product has highlighted the importance of application frequency as a variable during pre-clinical microbicide development, particularly in animal model studies. Methods To evaluate an association between application frequency and N-9 toxicity, experiments were performed using a mouse model of cervicovaginal microbicide safety. In this model system, changes in cervical and vaginal epithelial integrity, cytokine release, and immune cell infiltration were assessed after single and multiple exposures to N-9. Results After the initial application of N-9 (aqueous, 1%), considerable damage to the cervical epithelium (but not the vaginal epithelium) was observed as early as 10 min post-exposure and up to 8 h post-exposure. Subsequent daily exposures (up to 4 days) were characterized by diminished cervical toxicity relative to single exposures of like duration. Levels of pro-inflammatory cytokines released into the cervicovaginal lumen and the degree of CD14-positive immune cell infiltration proximal to the cervical epithelium were also dependent on the number of N-9 exposures. Conclusions Rather than causing cumulative cervical epithelial damage, repeated applications of N-9 were characterized by decreased sensitivity to N-9-associated toxicity and lower levels of immune cell recruitment. These results provide new insights into the failure of N-9-based microbicides and illustrate the importance of considering multiple exposure protocols in pre-clinical microbicide development strategies.

2012-01-01

37

Phenotypic and Functional Characterization of Vaginal Dendritic Cells in a Rat Model of Candida albicans Vaginitis  

PubMed Central

This study analyzes the phenotype of vaginal dendritic cells (VDCs), their antigenic presentation and activation of T-cell cytokine secretion, and their protective role in a rat model of Candida vaginitis. Histological observation demonstrated a significant accumulation of OX62+ VDCs in the mucosal epithelium of Candida albicans-infected rats at the third round of infection. We identified two subsets of OX62+ VDCs differing in the expression of CD4 molecule in both noninfected and Candida-infected rats. The OX62+ CD4+ subset of VDCs displayed a lymphoid cell-like morphology and expressed the T-cell antigen CD5, whereas the OX62+ CD4? VDC subset exhibited a myeloid morphology and was CD5 negative. Candida infection resulted in VDC maturation with enhanced expression of CD80 and CD134L on both CD4+ and CD4? VDC subsets at 2 and 6 weeks after Candida infection. CD5? CD4? CD86? CD80? CD134L+ VDCs from infected, but not noninfected, rats spontaneously released large amounts of interleukin-12 (IL-12) and tumor necrosis factor alpha, whereas all VDC subsets released comparable levels of IL-10 and IL-2 cytokines. Furthermore, OX62+ VDCs from infected rats primed naïve CD4+ T-cell proliferation and release of cytokines, including gamma interferon, IL-2, IL-6, and IL-10, in response to staphylococcal enterotoxin B stimulation in vitro. Adoptive transfer of highly purified OX62+ VDCs from infected rats induced a significant acceleration of fungal clearance compared with that in rats receiving naive VDCs, suggesting a protective role of VDCs in the anti-Candida mucosal immunity. Finally, VDC-mediated protection was associated with their ability to rapidly migrate to the vaginal mucosa and lymph nodes, as assessed by adoptive transfer of OX62+ VDCs labeled with 5 (and 6-)-carboxyfluorescein diacetate succinimidyl ester.

De Bernardis, Flavia; Lucciarini, Roberta; Boccanera, Maria; Amantini, Consuelo; Arancia, Silvia; Morrone, Stefania; Mosca, Michela; Cassone, Antonio; Santoni, Giorgio

2006-01-01

38

Phenotypic and functional characterization of vaginal dendritic cells in a rat model of Candida albicans vaginitis.  

PubMed

This study analyzes the phenotype of vaginal dendritic cells (VDCs), their antigenic presentation and activation of T-cell cytokine secretion, and their protective role in a rat model of Candida vaginitis. Histological observation demonstrated a significant accumulation of OX62(+) VDCs in the mucosal epithelium of Candida albicans-infected rats at the third round of infection. We identified two subsets of OX62(+) VDCs differing in the expression of CD4 molecule in both noninfected and Candida-infected rats. The OX62(+) CD4(+) subset of VDCs displayed a lymphoid cell-like morphology and expressed the T-cell antigen CD5, whereas the OX62(+) CD4(-) VDC subset exhibited a myeloid morphology and was CD5 negative. Candida infection resulted in VDC maturation with enhanced expression of CD80 and CD134L on both CD4(+) and CD4(-) VDC subsets at 2 and 6 weeks after Candida infection. CD5(-) CD4(-) CD86(-) CD80(-) CD134L(+) VDCs from infected, but not noninfected, rats spontaneously released large amounts of interleukin-12 (IL-12) and tumor necrosis factor alpha, whereas all VDC subsets released comparable levels of IL-10 and IL-2 cytokines. Furthermore, OX62(+) VDCs from infected rats primed naïve CD4(+) T-cell proliferation and release of cytokines, including gamma interferon, IL-2, IL-6, and IL-10, in response to staphylococcal enterotoxin B stimulation in vitro. Adoptive transfer of highly purified OX62(+) VDCs from infected rats induced a significant acceleration of fungal clearance compared with that in rats receiving naive VDCs, suggesting a protective role of VDCs in the anti-Candida mucosal immunity. Finally, VDC-mediated protection was associated with their ability to rapidly migrate to the vaginal mucosa and lymph nodes, as assessed by adoptive transfer of OX62(+) VDCs labeled with 5 (and 6-)-carboxyfluorescein diacetate succinimidyl ester. PMID:16790803

De Bernardis, Flavia; Lucciarini, Roberta; Boccanera, Maria; Amantini, Consuelo; Arancia, Silvia; Morrone, Stefania; Mosca, Michela; Cassone, Antonio; Santoni, Giorgio

2006-07-01

39

Lung Epithelial Stem Cells  

Microsoft Academic Search

\\u000a The lung epithelium is structurally and functionally a complex tissue composed of different cell types. It is exposed to toxic\\u000a agents and pathogens that can with time result in various lung diseases, including lung cancer. The major cell types in the\\u000a proximal tracheobronchial part are basal cells, goblet cells, ciliated cells, and cells of the submucosal glands. Further\\u000a down the

Magnus Karl Magnusson; Olafur Baldursson; Thorarinn Gudjonsson

40

Annexin-A1 identified as the oral epithelial cell anti-Candida effector moiety  

PubMed Central

Summary Innate and adaptive immunity are considered critical to protection against mucosal candidal infections. Among innate anti-Candida mechanisms, oral and vaginal epithelial cells have antifungal activity. The mechanism is fungistatic, acid-labile, and includes a requirement for cell contact by intact, but not necessarily live, epithelial cells. The purpose of this study was to use the acid-labile property to further characterize the effector moiety. Surface material extracted from PBS-, but not acid-treated epithelial cells, significantly inhibited the growth of Candida blastoconidia in a dose-dependent manner which was abrogated by prior heat and protease treatment. Proteins extracted from PBS-treated cells bound blastoconidia and hyphae more intensely than those from acid-treated cells. Proteins from PBS-treated cells eluted from Candida revealed two unique bands of approximately 33 and 45 kDa compared to acid-treated cells. Mass spectrometry identified these proteins as Annexin-A1 and actin, respectively. Oral epithelial cells stained positive for Annexin-A1, but not actin. Western blots showed reduced Annexin-A1 in proteins from acid-treated epithelial cells compared to those from PBS-treated epithelial cells. Lastly, it was demonstrated that immunoprecipitation of Annexin-A1 from proteins extracted from PBS-treated oral epithelial cells results in abrogation of inhibitory activity. Taken together, these results indicate that Annexin-A1 is a strong candidate for the epithelial cell anti-Candida effector protein.

Lilly, Elizabeth A.; Yano, Junko; Fidel, Paul L.

2010-01-01

41

Immune Cell-Mediated Protection against Vaginal Candidiasis: Evidence for a Major Role of Vaginal CD4+ T Cells and Possible Participation of Other Local Lymphocyte Effectors  

PubMed Central

The protective roles of different lymphocyte subsets were investigated in a rat vaginal candidiasis model by adoptive transfer of vaginal lymphocytes (VL) or sorted, purified CD3+ T cells, CD4+ or CD8+ T cells, or CD3? CD5+ B cells from the vaginas of naïve or immune rats following three rounds of Candida albicans infection. The adoptive transfer of total VL from nonimmune animals did not alter the course of vaginal candidiasis of the recipient rats. In contrast, the animals receiving total VL or CD3+ T cells from immune rats showed a highly significant acceleration of fungus clearance compared with animals which received nonimmune VL. The animals with vaginal CD3? CD5+ B cells transferred from immune rats also had fewer Candida CFU than the controls, but fungal clearance was significantly retarded with respect to the animals administered immune T cells. Sorted, purified CD4+ and CD8+ vaginal T cells from immune rats were also adoptively transferred to naïve animals. Although both populations were seen to accelerate the clearance of the fungus from the vagina, CD4+ T cells were much more effective than CD8+ T cells. Overall, there was no difference between the antifungal effects of immune vaginal CD4+ T cells and those achievable with the transfer of whole, immune VL. Histological observations of the vaginal tissues of rats with adoptively transferred immune T cells demonstrated a remarkable accumulation of lymphocytes in the subepithelial lamina propria and also infiltrating the mucosal epithelium. These results strongly suggest that distinct vaginal lymphocyte subsets participate in the adaptive anti-Candida immunity at the vaginal level, with the vaginal CD4+ T cells probably playing a major role.

Santoni, Giorgio; Boccanera, Maria; Adriani, Daniela; Lucciarini, Roberta; Amantini, Consuelo; Morrone, Stefania; Cassone, Antonio; De Bernardis, Flavia

2002-01-01

42

Immune cell-mediated protection against vaginal candidiasis: evidence for a major role of vaginal CD4(+) T cells and possible participation of other local lymphocyte effectors.  

PubMed

The protective roles of different lymphocyte subsets were investigated in a rat vaginal candidiasis model by adoptive transfer of vaginal lymphocytes (VL) or sorted, purified CD3(+) T cells, CD4(+) or CD8(+) T cells, or CD3(-) CD5(+) B cells from the vaginas of naïve or immune rats following three rounds of Candida albicans infection. The adoptive transfer of total VL from nonimmune animals did not alter the course of vaginal candidiasis of the recipient rats. In contrast, the animals receiving total VL or CD3(+) T cells from immune rats showed a highly significant acceleration of fungus clearance compared with animals which received nonimmune VL. The animals with vaginal CD3(-) CD5(+) B cells transferred from immune rats also had fewer Candida CFU than the controls, but fungal clearance was significantly retarded with respect to the animals administered immune T cells. Sorted, purified CD4(+) and CD8(+) vaginal T cells from immune rats were also adoptively transferred to naïve animals. Although both populations were seen to accelerate the clearance of the fungus from the vagina, CD4(+) T cells were much more effective than CD8(+) T cells. Overall, there was no difference between the antifungal effects of immune vaginal CD4(+) T cells and those achievable with the transfer of whole, immune VL. Histological observations of the vaginal tissues of rats with adoptively transferred immune T cells demonstrated a remarkable accumulation of lymphocytes in the subepithelial lamina propria and also infiltrating the mucosal epithelium. These results strongly suggest that distinct vaginal lymphocyte subsets participate in the adaptive anti-Candida immunity at the vaginal level, with the vaginal CD4(+) T cells probably playing a major role. PMID:12183521

Santoni, Giorgio; Boccanera, Maria; Adriani, Daniela; Lucciarini, Roberta; Amantini, Consuelo; Morrone, Stefania; Cassone, Antonio; De Bernardis, Flavia

2002-09-01

43

Protection of macaques from vaginal SHIV challenge by vaginally delivered inhibitors of virus-cell fusion  

Microsoft Academic Search

Human immunodeficiency virus type 1 (HIV-1) continues to spread, principally by heterosexual sex, but no vaccine is available. Hence, alternative prevention methods are needed to supplement educational and behavioural-modification programmes. One such approach is a vaginal microbicide: the application of inhibitory compounds before intercourse. Here, we have evaluated the microbicide concept using the rhesus macaque `high dose' vaginal transmission model

Ronald S. Veazey; Per Johan Klasse; Susan M. Schader; Qinxue Hu; Thomas J. Ketas; Min Lu; Preston A. Marx; Jason Dufour; Richard J. Colonno; Robin J. Shattock; Martin S. Springer; John P. Moore

2005-01-01

44

Clara epithelial cell depletion in the lung.  

PubMed

The bronchial epithelium has been increasingly recognized as an important immunomodulatory compartment in asthma and other lung diseases. Clara cells, which comprise the nonciliated secretory epithelial cells, are an important epithelial cell type with functions in the regulation of lung homeostasis and inflammation. Using naphthalene, Clara cells can be depleted within 24 h and regenerate by 1 month, hence, providing an easy method to study the impact of Clara cells on lung inflammation. PMID:23943457

Sonar, Sanchaita S; Dudda, Jan C

2013-01-01

45

Adhesion of Tritrichomonas foetus to Bovine Vaginal  

Microsoft Academic Search

An in vitro culture system of bovine vaginal epithelial cells (BVECs) was developed to study the cytopatho- genic effects of Tritrichomonas foetus and the role of lipophosphoglycan (LPG)-like cell surface glycoconjugates in adhesion of parasites to host cells. Exposure of BVEC monolayers to T. foetus resulted in extensive damage of monolayers. Host cell disruption was measured quantitatively by a trypan

Epithelial Cells; B. N. SINGH; J. J. LUCAS; D. H. BEACH; S. T. SHIN; R. O. GILBERT

1999-01-01

46

Epithelial Cell-Extracellular Matrix Interactions and Stem Cells in Airway Epithelial Regeneration  

Microsoft Academic Search

injured,leading to theimpairmentofits defensefunctions.Therapid restoration of the epithelial barrier is crucial for these patients. The complete regeneration of the airway epithelium is a complex phe- nomenon, including not only the epithelial wound repair but also theepithelialdifferentiationtoreconstituteafullywelldifferentiated and functional epithelium. The regeneration implies two partners: the epithelial stem\\/progenitor cells and factors able to regulate this process. Among these factors, epithelial cells-extracellular

Christelle Coraux; Jacqueline Roux; Thomas Jolly; Philippe Birembaut

2008-01-01

47

Patterning Bacterial Communities on Epithelial Cells  

PubMed Central

Micropatterning of bacteria using aqueous two phase system (ATPS) enables the localized culture and formation of physically separated bacterial communities on human epithelial cell sheets. This method was used to compare the effects of Escherichia coli strain MG1655 and an isogenic invasive counterpart that expresses the invasin (inv) gene from Yersinia pseudotuberculosis on the underlying epithelial cell layer. Large portions of the cell layer beneath the invasive strain were killed or detached while the non-invasive E. coli had no apparent effect on the epithelial cell layer over a 24 h observation period. In addition, simultaneous testing of the localized effects of three different bacterial species; E. coli MG1655, Shigella boydii KACC 10792 and Pseudomonas sp DSM 50906 on an epithelial cell layer is also demonstrated. The paper further shows the ability to use a bacterial predator, Bdellovibriobacteriovorus HD 100, to selectively remove the E. coli, S. boydii and P. sp communities from this bacteria-patterned epithelial cell layer. Importantly, predation and removal of the P. Sp was critical for maintaining viability of the underlying epithelial cells. Although this paper focuses on a few specific cell types, the technique should be broadly applicable to understand a variety of bacteria-epithelial cell interactions.

Dwidar, Mohammed; Leung, Brendan M.; Yaguchi, Toshiyuki; Takayama, Shuichi; Mitchell, Robert J.

2013-01-01

48

Differentiation of cultured epithelial cells: response to toxic agents.  

PubMed Central

Cell culture systems are instrumental in elucidating regulation of normal function and mechanisms of its perturbation by toxic substances. To this end, three applications of epithelial cells cultured with 3T3 feeder layer support are described. First, treatment of the premalignant human epidermal keratinocyte line SCC-12F2 with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate suppressed cell growth and differentiation. This agent produced a biphasic growth response greatly inhibiting cell growth at 1 to 10 nM, but much less above 100 nM. Expression of the differentiated functions involucrin and transglutaminase was found to be inhibited markedly at concentrations above 10 nM. Second, 3-methylcholanthrene toxicity was surveyed in a variety of rat epithelial cell types. The two most sensitive to growth inhibition were epidermal and mammary epithelial cells, while those from bladder, prostate, thyroid, and endometrium were insensitive to growth inhibition. Great differences were evident even among those cells derived from stratified squamous epithelia (epidermal, esophageal, vaginal, forestomach) despite their expression of aryl hydrocarbon hydroxylase activities to similar degrees. Finally, expression of estrogen receptors in rat endometrial cells was shown to be stimulated by the cAMP-elevating agent forskolin. Maximal stimulation of 3- to 6-fold occurred in 6 hr, compatible with a requirement for protein synthesis. Although expressing keratinocyte character (transglutaminase activity and envelope forming ability), the cells thus retain some hormonal character that may be modulated by cAMP-dependent kinase activity. Pursuit of such results will aid in understanding differences in response among cell types and species, in elucidating mechanisms of action of known toxic substances and, ultimately, in predicting toxicity of less well understood agents. Images FIGURE 1. FIGURE 4.

Rice, R H; LaMontagne, A D; Petito, C T; Rong, X H

1989-01-01

49

Symmetry breaking mechanism for epithelial cell polarization  

NASA Astrophysics Data System (ADS)

In multicellular organisms, epithelial cells form layers separating compartments responsible for different physiological functions. At the early stage of epithelial layer formation, each cell of an aggregate defines an inner and an outer side by breaking the symmetry of its initial state, in a process known as epithelial polarization. By integrating recent biochemical and biophysical data with stochastic simulations of the relevant reaction-diffusion system, we provide evidence that epithelial cell polarization is a chemical phase-separation process induced by a local bistability in the signaling network at the level of the cell membrane. The early symmetry breaking event triggering phase separation is induced by adhesion-dependent mechanical forces localized in the point of convergence of cell surfaces when a threshold number of confluent cells is reached. The generality of the emerging phase-separation scenario is likely common to many processes of cell polarity formation.

Veglio, A.; Gamba, A.; Nicodemi, M.; Bussolino, F.; Serini, G.

2009-09-01

50

Human glomerular epithelial cell proteoglycans  

SciTech Connect

Proteoglycans synthesized by cultures of human glomerular epithelial cells have been isolated and characterized. Three types of heparan sulfate were detected. Heparan sulfate proteoglycan I (HSPG-I; Kav 6B 0.04) was found in the cell layer and medium and accounted for 12% of the total proteoglycans synthesized. HSPG-II (Kav 6B 0.25) accounted for 18% of the proteoglycans and was located in the medium and cell layer. A third population (9% of the proteoglycan population), heparan sulfate glycosaminoglycan (HS-GAG; Kav 6B 0.4-0.8), had properties consistent with single glycosaminoglycan chains or their fragments and was found only in the cell layer. HSPG-I and HSPG-II from the cell layer had hydrophobic properties; they were released from the cell layer by mild trypsin treatment. HS-GAG lacked these properties, consisted of low-molecular-mass heparan sulfate oligosaccharides, and were intracellular. HSPG-I and -II released to the medium lacked hydrophobic properties. The cells also produced three distinct types of chondroitin sulfates. The major species, chondroitin sulfate proteoglycan I (CSPG-I) eluted in the excluded volume of a Sepharose CL-6B column, accounted for 30% of the proteoglycans detected, and was found in both the cell layer and medium. Cell layer CSPG-I bound to octyl-Sepharose. It was released from the cell layer by mild trypsin treatment. CSPG-II (Kav 6B 0.1-0.23) accounted for 10% of the total 35S-labeled macromolecules and was found predominantly in the culture medium. A small amount of CS-GAG (Kav 6B 0.25-0.6) is present in the cell extract and like HS-GAG is intracellular. Pulse-chase experiments indicated that HSPG-I and -II and CSPG-I and -II are lost from the cell layer either by direct release into the medium or by internalization where they are metabolized to single glycosaminoglycan chains and subsequently to inorganic sulfate.

Thomas, G.J.; Jenner, L.; Mason, R.M.; Davies, M. (Univ. of Wales College of Medicine, Royal Infirmary, Cardiff (England))

1990-04-01

51

Airway epithelial cell responses to ozone injury  

SciTech Connect

The airway epithelial cell is an important target in ozone injury. Once activated, the airway epithelium responds in three phases. The initial, or immediate phase, involves activation of constitutive cells, often through direct covalent interactions including the formation of secondary ozonolysis products-hydroxyhydroperoxides, aldehydes, and hydrogen peroxide. Recently, we found hydroxyhydroperoxides to be potent agonists; of bioactive eicosanoid formation by human airway epithelial cells in culture. Other probable immediate events include activation and inactivation of enzymes present on the epithelial surface (e.g., neutral endopeptidase). During the next 2 to 24 hr, or early phase, epithelial cells respond by synthesis and release of chemotactic factors, including chemokines-macrophage inflammatory protein-2, RANTES, and interleukin-8. Infiltrating leukocytes during this period also release elastase, an important agonist of epithelial cell mucus secretion and additional chemokine formation. The third (late) phase of ozone injury is characterized by eosinophil or monocyte infiltration. Cytokine expression leads to alteration of structural protein synthesis, with increases in fibronectin evident by in situ hybridization. Synthesis of epithelial antiproteases, e.g., secretary leukocyte protease inhibitor, may also increase locally 24 to 48 hr after elastase concentrations become excessive. Thus, the epithelium is not merely a passive barrier to ozone injury but has a dynamic role in directing the migration, activating, and then counteracting inflammatory cells. Through these complex interactions, epithelial cells can be viewed as the initiators (alpha) and the receptors (omega) of ozone-induced airway disease. 51 refs., 2 figs., 3 tabs.

Leikauf, G.D.; Simpson, L.G.; Zhao, Qiyu [Univ. of Cincinnati Medical Center, OH (United States)] [and others

1995-03-01

52

Induction of avian ?-defensins by CpG oligodeoxynucleotides and proinflammatory cytokines in hen vaginal cells in vitro.  

PubMed

Immune function in the vagina of hen oviduct is essential to prevent infection by microorganisms colonizing in the cloaca. The aim of this study was to determine whether CpG oligodeoxynucleotides (CpG-ODN) stimulate the expression of avian ?-defensins (AvBDs) in hen vaginal cells. Specific questions were whether CpG-ODN affects the expression of AvBDs and proinflammatory cytokines and whether the cytokines affect AvBDs expression in vaginal cells. The dispersed vaginal cells of White Leghorn laying hens were cultured and stimulated by different doses of lipopolysaccharide (LPS), CpG-ODN, interleukin 1? (IL1B), or IL6. The cultured cell population contained epithelial cells, fibroblast-like cells, and CD45-positive leukocytes. The immunoreactive AvBD3, -10, and -12 were localized in the mucosal epithelium in the section of the vagina. The expression of AvBDs, IL1B, and IL6 was analyzed by quantitative RT-PCR. RT-PCR analysis showed the expression of AvBD1, -3, -4, -5, -10, and -12 in the cultured vaginal cells without stimulation. Toll-like receptors (TLRs) 4 and 21, which recognize LPS and CpG-ODN respectively and IL1 and IL6 receptors (IL1R1 and IL6R) were also expressed in them. The expression of IL1B, IL6, and AvBD10 and -12 was upregulated by LPS, whereas only IL1B and IL6 were upregulated by CpG-ODN. IL1B stimulation upregulated AvBD1 and -3 expression, whereas IL6 stimulation did not cause changes in AvBDs expression. These results suggest that CpG-ODN derived from microbes upregulates the expression of IL1B and IL6 by interaction with TLR21 and then IL1B induces AvBD1 and -3 to prevent infection in the vagina. PMID:23625580

Sonoda, Yuka; Abdel Mageed, Ahmad M; Isobe, Naoki; Yoshimura, Yukinori

2013-05-21

53

A Population-Based Study of Squamous Cell Vaginal Cancer: HPV and Cofactors  

Microsoft Academic Search

Background. Little is known about the etiology of in situ or invasive squamous cell cancer of the vagina. It is thought that some vaginal cancers may have the same etiology as cervical cancer. It is also not known whether in situ and invasive vaginal cancer share the same etiologic factors. We conducted a study to evaluate risk factors for in

Janet R. Daling; Margaret M. Madeleine; Stephen M. Schwartz; Katherine A. Shera; Joseph J. Carter; Barbara McKnight; Peggy L. Porter; Denise A. Galloway; James K. McDougall; Hisham Tamimi

2002-01-01

54

Propagation and Immortalization of Human Lens Epithelial Cells in Culture  

Microsoft Academic Search

Purpose. To establish primary and immortalized cell cultures of human lens epithelial cells for a model system investigating human lens epithelial physiology and cataract. Methods. Human lens epithelial cells in culture were grown by isolating epithelium fragments from infant human lenses from patients who underwent treatment for retinopathy of prema- turity and by allowing epithelial cells to grow from explants.

Usha P. Andley; Johng S. Rhim; Leo T. Chylack; Timothy P. Fleming

55

Human papillomavirus in vulvar and vaginal carcinoma cell lines.  

PubMed Central

A number of reports associate human papillomavirus (HPV) with cervical cancer and cancer cell lines derived from this tumour type. Considerably fewer reports have focused on the role of HPV in carcinomas from other sites of female anogenital squamous epithelia. In this study we have tested for the presence of HPV in eight low-passage vulvar carcinoma cell lines and one extensively passaged cell line, A431. One cell line from a primary vaginal carcinoma was included. The presence of the HPV was evaluated by the polymerase chain reaction (PCR), by Southern blot analysis and by two-dimensional gel electrophoresis. General primer-mediated PCR was applied by using primers from the L1 region, E1 region and HPV 16 E7 region. Southern blot hybridisation was performed under low-stringency conditions (Tm = -35 degrees C) using a whole genomic HPV 6/16/18 probe mixture and under high stringency conditions (Tm = -18 degrees C) with the whole genomic probes of HPV 16 and 33. HPV 16 E6-E7 mRNA was assessed by ribonuclease protection assay (RPA). HPV was found in only one vulvar carcinoma cell line, UM-SCV-6. The identified type, HPV 16, was integrated in the cell genome and could be amplified with all primers used. Also E6-E7 transcripts were found in these cells. Five original tumour biopsies were available from the HPV-negative cell lines for in situ hybridisation. All these were HPV negative with both the HPV 6/16/18 screening probe mixture under low stringency and the HPV 16 probe under high stringency. The results indicate that vulvar carcinoma cell lines contain HPV less frequently than cervical carcinoma cell lines and suggest that a significant proportion of vulvar carcinomas may evolve by an HPV-independent mechanism. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6

Hietanen, S.; Grenman, S.; Syrjanen, K.; Lappalainen, K.; Kauppinen, J.; Carey, T.; Syrjanen, S.

1995-01-01

56

Fusobacterium nucleatum Transports Noninvasive Streptococcus cristatus into Human Epithelial Cells  

Microsoft Academic Search

Analysis of human buccal epithelial cells frequently reveals an intracellular polymicrobial consortium of bacteria. Although several oral bacteria have been demonstrated to invade cultured epithelial cells, several others appear unable to internalize. We hypothesized that normally noninvasive bacteria may gain entry into epithelial cells via adhesion to invasive bacteria. Fusobacterium nucleatum is capable of binding to and invading oral epithelial

Andrew M. Edwards; Tracy J. Grossman; Joel D. Rudney

2006-01-01

57

Epithelial-mesenchymal transition of tubular epithelial cells in human renal biopsies  

Microsoft Academic Search

Epithelial-mesenchymal transition of tubular epithelial cells in human renal biopsies.BackgroundIn recent studies performed on cultured cells and experimental nephropathies, it has been hypothesized that tubular epithelial cells (TEC), via epithelial-mesenchymal transformation (EMT), can become collagen-producing cells. According to this theory, they should proceed through several activating steps, such as proliferation and phenotype changes, to eventually synthesize extracellular matrix (ECM).MethodsTo evaluate

Maria P Rastaldi; Franco Ferrario; Laura Giardino; Giacomo Dell'Antonio; Carlo Grillo; Paolo Grillo; Frank Strutz; Gerhard A Müller; Giuliano Colasanti; Giuseppe D'Amico

2002-01-01

58

HIV Infection in Gastric Epithelial Cells.  

PubMed

Many chronic human immunodeficiency virus (HIV) patients suffer from gastric complaints, including gastric tuberculosis and coinfection of other pathogens. Recent work has demonstrated that a variety of nonimmune cells can act as viral reservoirs, even at the early stage of HIV infection. In this study, we detect HIV viral particles, proteins, and nucleic acids in gastric epithelial cells using clinical samples. These observations are further supported by a simian immunodeficiency virus-infected macaque model. Further, the number of HIV-infected gastric epithelial cells is positively associated with blood viral load, and is negatively correlated with CD4 lymphocyte cell counts. We also demonstrate that HIV infection is accompanied by severe inflammatory response in gastric mucosa. Additionally, HIV infection activates signal transducer and activator of transcription 3 and RelA, and enhances the production of interleukin 6 and tumor necrosis factor ? in gastric epithelial cells. The present data suggest that the gastric epithelial cells are natural targets of HIV infection, and HIV infection in epithelial cells contributes to HIV-induced gastric mucosal inflammation. PMID:23852124

Liu, Rui; Huang, Lei; Li, Jingyi; Zhou, Xianzhi; Zhang, Haiyuan; Zhang, Tao; Lei, Yunlong; Wang, Kui; Xie, Na; Zheng, Yongtang; Wang, Fusheng; Nice, Edouard C; Rong, Lijun; Huang, Canhua; Wei, Yuquan

2013-07-11

59

Intrinsic ageing of gut epithelial stem cells.  

PubMed

The maintenance of integrity of gut epithelium is essential for the well being of the organism throughout life. Gut epithelium is maintained through a carefully controlled balance between cell loss and renewal, which is sustained by the proliferation of the epithelial stem cells. Although these stem cells show no intrinsic limit to their proliferative capacity, recent evidence indicates that they suffer important functional impairments during the course of ageing. This paper reviews what is currently known about intrinsic ageing of gut epithelial stem cells and its functional consequences. PMID:15563938

Kirkwood, Thomas B L

2004-12-01

60

Regulation of intestinal epithelial cells transcriptome by enteric glial cells: impact on intestinal epithelial barrier functions  

Microsoft Academic Search

BACKGROUND: Emerging evidences suggest that enteric glial cells (EGC), a major constituent of the enteric nervous system (ENS), are key regulators of intestinal epithelial barrier (IEB) functions. Indeed EGC inhibit intestinal epithelial cells (IEC) proliferation and increase IEB paracellular permeability. However, the role of EGC on other important barrier functions and the signalling pathways involved in their effects are currently

Laurianne Van Landeghem; Maxime M Mahé; Raluca Teusan; Jean Léger; Isabelle Guisle; Rémi Houlgatte; Michel Neunlist

2009-01-01

61

What Is Vaginal Cancer?  

MedlinePLUS

... cancer There are several types of vaginal cancer. Squamous cell carcinoma About 70 of every 100 cases of vaginal cancer are squamous cell carcinomas . These cancers begin in the squamous cells that ...

62

Reciprocal interference between Lactobacillus spp. and Gardnerella vaginalis on initial adherence to epithelial cells.  

PubMed

Bacterial vaginosis (BV) is the most common vaginal disorder in women of child-bearing age. It is widely accepted that the microbial switch from normal microflora to the flora commonly associated with BV is characterized by a decrease in vaginal colonization by specific Lactobacillus species together with an increase of G. vaginalis and other anaerobes. However, the order of events leading to the development of BV remains poorly characterized and it is unclear whether the decrease in lactobacilli is a cause or a consequence of the increase in the population density of anaerobes. Our goal was to characterize the interaction between two Gardnerella vaginalis strains, one of which was isolated from a healthy woman (strain 5-1) and the other from a woman diagnosed with BV (strain 101), and vaginal lactobacilli on the adherence to cervical epithelial cells. In order to simulate the transition from vaginal health to BV, the lactobacilli were cultured with the epithelial cells first, and then the G. vaginalis strain was introduced. We quantified the inhibition of G. vaginalis adherence by the lactobacilli and displacement of adherent lactobacilli by G. vaginalis. Our results confirmed that pathogenic G vaginalis 101 had a higher capacity for adhesion to the cervical epithelial cells than strain 5-1. Interestingly, strain 101 displaced L. crispatus but not L. iners whereas strain 5-1 had less of an effect and did not affect the two species differently. Furthermore, L. iners actually enhanced adhesion of strain 101 but not of strain 5-1. These results suggest that BV-causing G. vaginalis and L. iners do not interfere with one another, which may help to explain previous reports that women who are colonized with L. iners are more likely to develop BV. PMID:23935396

Castro, Joana; Henriques, Ana; Machado, António; Henriques, Mariana; Jefferson, Kimberly K; Cerca, Nuno

2013-07-20

63

Reciprocal Interference between Lactobacillus spp. and Gardnerella vaginalis on Initial Adherence to Epithelial Cells  

PubMed Central

Bacterial vaginosis (BV) is the most common vaginal disorder in women of child-bearing age. It is widely accepted that the microbial switch from normal microflora to the flora commonly associated with BV is characterized by a decrease in vaginal colonization by specific Lactobacillus species together with an increase of G. vaginalis and other anaerobes. However, the order of events leading to the development of BV remains poorly characterized and it is unclear whether the decrease in lactobacilli is a cause or a consequence of the increase in the population density of anaerobes. Our goal was to characterize the interaction between two Gardnerella vaginalis strains, one of which was isolated from a healthy woman (strain 5-1) and the other from a woman diagnosed with BV (strain 101), and vaginal lactobacilli on the adherence to cervical epithelial cells. In order to simulate the transition from vaginal health to BV, the lactobacilli were cultured with the epithelial cells first, and then the G. vaginalis strain was introduced. We quantified the inhibition of G. vaginalis adherence by the lactobacilli and displacement of adherent lactobacilli by G. vaginalis. Our results confirmed that pathogenic G vaginalis 101 had a higher capacity for adhesion to the cervical epithelial cells than strain 5-1. Interestingly, strain 101 displaced L. crispatus but not L. iners whereas strain 5-1 had less of an effect and did not affect the two species differently. Furthermore, L. iners actually enhanced adhesion of strain 101 but not of strain 5-1. These results suggest that BV-causing G. vaginalis and L. iners do not interfere with one another, which may help to explain previous reports that women who are colonized with L. iners are more likely to develop BV.

Castro, Joana; Henriques, Ana; Machado, Antonio; Henriques, Mariana; Jefferson, Kimberly K.; Cerca, Nuno

2013-01-01

64

Induced Pluripotency of Human Prostatic Epithelial Cells  

PubMed Central

Induced pluripotent stem (iPS) cells are a valuable resource for discovery of epigenetic changes critical to cell type-specific differentiation. Although iPS cells have been generated from other terminally differentiated cells, the reprogramming of normal adult human basal prostatic epithelial (E-PZ) cells to a pluripotent state has not been reported. Here, we attempted to reprogram E-PZ cells by forced expression of Oct4, Sox2, c-Myc, and Klf4 using lentiviral vectors and obtained embryonic stem cell (ESC)-like colonies at a frequency of 0.01%. These E-PZ-iPS-like cells with normal karyotype gained expression of pluripotent genes typical of iPS cells (Tra-1-81, SSEA-3, Nanog, Sox2, and Oct4) and lost gene expression characteristic of basal prostatic epithelial cells (CK5, CK14, and p63). E-PZ-iPS-like cells demonstrated pluripotency by differentiating into ectodermal, mesodermal, and endodermal cells in vitro, although lack of teratoma formation in vivo and incomplete demethylation of pluripotency genes suggested only partial reprogramming. Importantly, E-PZ-iPS-like cells re-expressed basal epithelial cell markers (CD44, p63, MAO-A) in response to prostate-specific medium in spheroid culture. Androgen induced expression of androgen receptor (AR), and co-culture with rat urogenital sinus further induced expression of prostate-specific antigen (PSA), a hallmark of secretory cells, suggesting that E-PZ-iPS-like cells have the capacity to differentiate into prostatic basal and secretory epithelial cells. Finally, when injected into mice, E-PZ-iPS-like cells expressed basal epithelial cell markers including CD44 and p63. When co-injected with rat urogenital mesenchyme, E-PZ-iPS-like cells expressed AR and expression of p63 and CD44 was repressed. DNA methylation profiling identified epigenetic changes in key pathways and genes involved in prostatic differentiation as E-PZ-iPS-like cells converted to differentiated AR- and PSA-expressing cells. Our results suggest that iPS-like cells derived from prostatic epithelial cells are pluripotent and capable of prostatic differentiation; therefore, provide a novel model for investigating epigenetic changes involved in prostate cell lineage specification.

Zhao, Hongjuan; Sun, Ning; Young, Sarah R.; Nolley, Rosalie; Santos, Jennifer; Wu, Joseph C.; Peehl, Donna M.

2013-01-01

65

Wound healing of intestinal epithelial cells  

PubMed Central

The intestinal epithelial cells (IECs) form a selective permeability barrier separating luminal content from underlying tissues. Upon injury, the intestinal epithelium undergoes a wound healing process. Intestinal wound healing is dependent on the balance of three cellular events; restitution, proliferation, and differentiation of epithelial cells adjacent to the wounded area. Previous studies have shown that various regulatory peptides, including growth factors and cytokines, modulate intestinal epithelial wound healing. Recent studies have revealed that novel factors, which include toll-like receptors (TLRs), regulatory peptides, particular dietary factors, and some gastroprotective agents, also modulate intestinal epithelial wound repair. Among these factors, the activation of TLRs by commensal bacteria is suggested to play an essential role in the maintenance of gut homeostasis. Recent studies suggest that mutations and dysregulation of TLRs could be major contributing factors in the predisposition and perpetuation of inflammatory bowel disease. Additionally, studies have shown that specific signaling pathways are involved in IEC wound repair. In this review, we summarize the function of IECs, the process of intestinal epithelial wound healing, and the functions and mechanisms of the various factors that contribute to gut homeostasis and intestinal epithelial wound healing.

Iizuka, Masahiro; Konno, Shiho

2011-01-01

66

Wnt Proteins in Mammary Epithelial Cell Transformation.  

National Technical Information Service (NTIS)

We have assessed the ability of Wnt-1, Wnt-2, Wnt-3, Wnt-3A, Wnt-4, Wnt-5A, Wnt-5B, Wnt- 6, Wnt-7A, and Wnt-7B to transform mammary epithelial cells. Epitope-tagged Wnt proteins were expressed in cells and the proteins were analyzed by immunoblots. Bxtrac...

J. Kitajewski M. Julius Z. Zheng

1995-01-01

67

Expression and gene transcript of Fc receptors for IgG, HLA class II antigens and Langerhans cells in human cervico-vaginal epithelium.  

PubMed Central

The mechanism of transmission of HIV from the male to the female genital tract or in the reverse order is not clear. CD4 glycoprotein is the receptor for HIV and Langerhans cells and the related dendritic cells could play a role in the initial transmission of HIV. Fc receptors (FcR) for IgG might be involved in antibody-mediated binding of HIV. We carried out an immunohistological study of normal human cervical and vaginal epithelia for the presence of CD4 glycoprotein, Langerhans cells and FcR to IgG. CD4+ glycoprotein was not found in the vaginal or cervical epithelium, with the exception of a few endocervical epithelial cells. A small number of CD4+ mononuclear cells were found in the endocervical epithelium of a third of the specimens but a large number of CD4+ cells was found in the submucosa of most of the cervical and vaginal specimens. Langerhans cells expressing CD4, HLA class II, Fc gamma R2 and Fc gamma R3 were detected in most vaginal, ectocervical and transformation zone epithelia and in 9/14 endocervical tissues. Fc gamma R3 was detected in about two-thirds of the columnar endocervical epithelium and the transformation zone. A smaller number of specimens expressed Fc gamma R2 in these epithelia, but Fc gamma R1 was not detected. We then demonstrated mRNA for Fc gamma R3 in the columnar endocervical epithelial cells and transformation zone by in situ hybridization, using a CD16-RNA probe. Fc gamma R3 and Fc gamma R2 gene transcripts were also found in fetal cervical tissue by applying the polymerase chain reaction to amplify portions of the Fc gamma R3 and Fc gamma R2 coding sequences in cDNA prepared from fetal RNA. HLA-DR was found in the endocervical cells, transformation zone and in Langerhans cells of all specimens. The presence of Langerhans cells, Fc gamma receptors and HLA class II antigen offers three potential mechanisms for cervico-vaginal HIV transmission: (i) direct HIV infection of Langerhans cells, (ii) binding of HIV antibody complexes to cervical epithelial Fc gamma receptors and (iii) binding of HIV infected CD4+ cells to cervical HLA class II antigen which may infect these or the adjacent CD4+ cells. Images Fig. 1 Fig. 2

Hussain, L A; Kelly, C G; Fellowes, R; Hecht, E M; Wilson, J; Chapman, M; Lehner, T

1992-01-01

68

Replication of cultured lung epithelial cells  

SciTech Connect

The authors have investigated the conditions necessary to support replication of lung type 2 epithelial cells in culture. Cells were isolated from mature fetal rabbit lungs (29d gestation) and cultured on feeder layers of mitotically inactivated 3T3 fibroblasts. The epithelial nature of the cells was demonstrated by indirect immunofluorescent staining for keratin and by polyacid dichrome stain. Ultrastructural examination during the first week showed that the cells contained myofilaments, microvilli and lamellar bodies (markers for type 2 cells). The following changes were observed after the first week: increase in cell size; loss of lamellar bodies and appearance of multivesicular bodies; increase in rough endoplasmic reticulum and golgi; increase in tonafilaments and well-defined junctions. General cell morphology was good for up to 10 wk. Cells cultured on plastic surface degenerated after 1 wk. Cell replication was assayed by autoradiography of cultures exposed to (/sup 3/H)-thymidine and by direct cell counts. The cells did not replicate during the first week; however, between 2-10 wk the cells incorporated the label and went through approximately 6 population doublings. They have demonstrated that lung alveolar epithelial cells can replicate in culture if they are maintained on an appropriate substrate. The coincidence of ability to replicate and loss of markers for differentiation may reflect the dichotomy between growth and differentiation commonly observed in developing systems.

Guzowski, D.; Bienkowski, R.

1986-03-05

69

Characterization of protocadherin-1 expression in primary bronchial epithelial cells: association with epithelial cell differentiation.  

PubMed

Protocadherin-1 (PCDH1) is a novel susceptibility gene for asthma that is expressed in airway epithelium. We aimed to characterize PCDH1 mRNA transcripts and protein expression in primary bronchial epithelial cells and to determine regulation of PCDH1 during mucociliary differentiation. Total RNA and protein were isolated from human primary bronchial epithelial cells. PCDH1 transcripts were characterized by rapid amplification of cDNA ends in bronchial epithelial cells of 4 subjects. PCDH1 expression was quantified by quantitative RT-PCR and Western blotting in bronchial epithelial cells directly ex vivo and after air liquid interface (ALI) or submerged culture. We identified 5 novel exons on the 5' end and 1 exon on the 3' end of PCDH1. Novel transcripts showed major variation in expression of intracellular conserved motifs. Expression levels of PCDH1 transcripts encoding exon 1-2 were 4-fold higher, and transcripts encoding exon 3-4 were 15-fold higher in freshly isolated bronchial epithelial cells than in submerged cultures. PCDH1 mRNA (3- to 8-fold) and protein levels (2- to 3-fold) were strongly up-regulated during mucociliary differentiation of primary bronchial epithelial cells in ALI cultures. In summary, PCDH1 transcripts display remarkable variability in expression of conserved intracellular signaling domains. Enhanced PCDH1 expression levels strongly correlate with differentiation of bronchial epithelial cells. PMID:21982948

Koning, Henk; Sayers, Ian; Stewart, Ceri E; de Jong, Debora; Ten Hacken, Nick H T; Postma, Dirkje S; van Oosterhout, Antoon J M; Nawijn, Martijn C; Koppelman, Gerard H

2011-10-07

70

Identification and Characterization of Bacterial Vaginosis-Associated Pathogens Using a Comprehensive Cervical-Vaginal Epithelial Coculture Assay  

PubMed Central

Bacterial vaginosis (BV) is the most commonly treated female reproductive tract affliction, characterized by the displacement of healthy lactobacilli by an overgrowth of pathogenic bacteria. BV can contribute to pathogenic inflammation, preterm birth, and susceptibility to sexually transmitted infections. As the bacteria responsible for BV pathogenicity and their interactions with host immunity are not understood, we sought to evaluate the effects of BV-associated bacteria on reproductive epithelia. Here we have characterized the interaction between BV-associated bacteria and the female reproductive tract by measuring cytokine and defensin induction in three types of FRT epithelial cells following bacterial inoculation. Four BV-associated bacteria were evaluated alongside six lactobacilli for a comparative assessment. While responses differed between epithelial cell types, our model showed good agreement with clinical BV trends. We observed a distinct cytokine and human ?-defensin 2 response to BV-associated bacteria, especially Atopobium vaginae, compared to most lactobacilli. One lactobacillus species, Lactobacillus vaginalis, induced an immune response similar to that elicited by BV-associated bacteria, stimulating significantly higher levels of cytokines and human ?-defensin 2 than other lactobacilli. These data provide an important prioritization of BV-associated bacteria and support further characterization of reproductive bacteria and their interactions with host epithelia. Additionally, they demonstrate the distinct immune response potentials of epithelial cells from different locations along the female reproductive tract.

Eade, Colleen R.; Diaz, Camila; Wood, Matthew P.; Anastos, Kathryn; Patterson, Bruce K.; Gupta, Phalguni; Cole, Amy L.; Cole, Alexander M.

2012-01-01

71

Nasal Epithelial Cells as Surrogates for Bronchial Epithelial Cells in Airway Inflammation Studies  

Microsoft Academic Search

and tissue inhibitor of metalloproteinase-1. Despite differences in absolute mediator levels, the responses of nasal and bronchial epithelial cells to cytokine stimulation were similar, expression of relevant surface receptors was comparable, and there were signifi- cantcorrelations betweennasaland bronchialcellmediatorrelease. Therefore, nasal epithelial cultures constitute an accessible surro- gate for studying lower airway inflammation.

Catherine M. McDougall; Morgan G. Blaylock; J. Graham Douglas; Richard J. Brooker; Peter J. Helms; Garry M. Walsh

2008-01-01

72

Control of the epithelial stem cell epigenome: the shaping of epithelial stem cell identity.  

PubMed

The squamous epithelium covering the skin and oral mucosa relies on epithelial stem cells for tissue renewal. Dynamic changes in DNA methylation, histone methylation and acetylation, and higher order chromatin structure are required to preserve their self-renewal capacity while orchestrating the timely execution of cell differentiation programs. This complex network of epigenetic modifications shapes the epithelial stem cell identity and fate. Pathological alterations can be perceived by aberrant chromatin sensors, such as the INK4/ARF locus, which initiate tumor suppressive cell senescence programs, and can often result in epithelial stem cell exhaustion. Unveiling the mechanisms controlling the epigenome in epithelial stem cells may help protect against the loss of their tissue regenerative capacity, thereby preventing premature aging without increasing cancer risk. PMID:23434069

Iglesias-Bartolome, Ramiro; Callejas-Valera, Juan Luis; Gutkind, J Silvio

2013-02-19

73

Molecular Aspects of Epithelial ?? T Cell Regulation  

PubMed Central

?? T cells lie at the interface between innate and adaptive immunity, sharing features with both arms of the immune system. The vast majority of ?? T cells reside in epithelial layers of tissues such as skin, gut, lung, tongue and reproductive tract. Here they provide a first line of defense against environmental attack. While the existence of epithelial resident ?? T cells has been known for over 20 years, understanding the molecular events regulating development and function of these cells is still in progress. Here we review recent advances in the field, with a particular emphasis on the ?? T cell population resident in mouse epidermis. These studies have enhanced our knowledge and understanding of the life cycle of this enigmatic population of cells.

Witherden, Deborah A; Havran, Wendy L

2011-01-01

74

Corneal Epithelial Stem Cells: Deficiency and Regulation  

Microsoft Academic Search

The corneal epithelium is continuously renewed by a population of stem cells that reside in the corneoscleral junction, otherwise\\u000a known as the limbus. These limbal epithelial stem cells (LESC) are imperative for corneal maintenance with deficiencies leading\\u000a to in-growth of conjunctival cells, neovascularisation of the corneal stroma and eventual corneal opacity and visual loss.\\u000a One such disease that has traditionally

Genevieve A. Secker; Julie T. Daniels

2008-01-01

75

Cell Cycle in Normal and Malignant Breast Epithelial Cells.  

National Technical Information Service (NTIS)

Breast cancer is a disease where breast epithelial cells become refractory to appropriate growth and differentiation signals. It is likely that numerous genetic changes can contribute to malignant transformation, including mutations that alter the cell cy...

1997-01-01

76

Vaginal Cancer  

MedlinePLUS

... body). There are four types of vaginal cancer: Squamous cell carcinoma. Squamous cell carcinoma is a type of skin cancer that begins in the cells lining the vagina, most often in the area closest to the cervix. Squamous cell cancer makes up 85% to 90% of ...

77

Cross-talk between intraepithelial ?? T cells and epithelial cells.  

PubMed

Intraepithelial ?? T cells play pivotal roles in homeostasis, tissue repair, inflammation, and protection from malignancy. In some tissues, ?? T cells are the only resident T cell population, whereas in others, they coexist with ?? T cells and other lymphocyte populations. ?? T cell function in the epithelium requires constant communication between cells in the form of cell-to-cell contacts and cell-to-matrix interactions. These interactions coordinate with the timely production of specific cytokines, chemokines, growth factors, and glycosaminoglycans, which have specialized effects on neighboring epithelial cells. Antigens that activate these T cells are not well-defined, and they do not express classic costimulatory or coreceptor molecules. As such, an understanding of the mechanisms used by epithelial ?? T cells to maintain homeostasis and facilitate wound repair has necessitated the identification of novel molecular interactions between ?? T cells and their neighboring epithelial cells. PMID:23620015

Witherden, Deborah A; Havran, Wendy L

2013-04-25

78

Method for Modulating Epithelial Stem Cell Lineage.  

National Technical Information Service (NTIS)

The present invention relates to methods of modulating epithelial stem cell lineage by regulating the expression of Lef1 or a BMP inhibitor and/or the stability of beta-catenin or the expression of a Wnt; regulating the expression or activity of GATA-3; o...

C. Jamora C. Kaufman E. Fuchs K. Kobielak

2004-01-01

79

Protons sensitize epithelial cells to mesenchymal transition.  

PubMed

Proton radiotherapy has gained more favor among oncologists as a treatment option for localized and deep-seated tumors. In addition, protons are a major constituent of the space radiation astronauts receive during space flights. The potential for these exposures to lead to, or enhance cancer risk has not been well studied. Our objective is to study the biological effects of low energy protons on epithelial cells and its propensity to enhance transforming growth factor beta 1 (TGF?1)-mediated epithelial-mesenchymal transition (EMT), a process occurring during tumor progression and critical for invasion and metastasis. Non-transformed mink lung epithelial cells (Mv1Lu) and hTERT- immortalized human esophageal epithelial cells (EPC) were used in this study. EMT was identified by alterations in cell morphology, EMT-related gene expression changes determined using real-time PCR, and EMT changes in specific cellular markers detected by immunostaining and western blotting. Although TGF?1 treatment alone is able to induce EMT in both Mv1Lu and EPC cells, low energy protons (5 MeV) at doses as low as 0.1 Gy can enhance TGF?1 induced EMT. Protons alone can also induce a mild induction of EMT. SD208, a potent TGF? Receptor 1 (TGF?R1) kinase inhibitor, can efficiently block TGF?1/Smad signaling and attenuate EMT induction. We suggest a model for EMT after proton irradiation in normal and cancerous tissue based on our results that showed that low and high doses of protons can sensitize normal human epithelial cells to mesenchymal transition, more prominently in the presence of TGF?1, but also in the absence of TGF?1. PMID:22844446

Wang, Minli; Hada, Megumi; Saha, Janapriya; Sridharan, Deepa M; Pluth, Janice M; Cucinotta, Francis A

2012-07-23

80

Streptococcus pneumoniae early response genes to human lung epithelial cells  

Microsoft Academic Search

BACKGROUND: Streptococcus pneumoniae infection starts from colonization of the host respiratory tract where interaction with host respiratory tract epithelial cells occurs. To investigate pneumococcal genes that are involved in the early stage of interaction with host epithelial cells, transcriptional responses of an encapsulated pathogenic pneumococcal strain TIGR4 upon exposure to human lung epithelial cells A549 for 0.5 h and 1

Xin-Ming Song; Wayne Connor; Karsten Hokamp; Lorne A Babiuk; Andrew A Potter

2008-01-01

81

Change in cell shape is required for matrix metalloproteinase-induced epithelial-mesenchymal transition of mammary epithelial cells  

Microsoft Academic Search

Cell morphology dictates response to a wide variety of stimuli, controlling cell metabolism, differentiation, proliferation, and death. Epithelial-mesenchymal transition (EMT) is a developmental process in which epithelial cells acquire migratory characteristics, and in the process convert from a 'cuboidal' epithelial structure into an elongated mesenchymal shape. We had shown previously that matrix metalloproteinase-3 (MMP3) can stimulate EMT of cultured mouse

Celeste M. Nelson; Davitte Khauv; Mina J. Bissell; Derek C. Radisky

2008-01-01

82

Th17 Cells and IL-17 in Protective Immunity to Vaginal Candidiasis  

PubMed Central

Background Th17 cells play a major role in coordinating the host defence in oropharyngeal candidiasis. In this study we investigated the involvement of the Th17 response in an animal model of vulvovaginal candidiasis (VVC). Methods To monitor the course of infection we exploited a new in vivo imaging technique. Results i) The progression of VVC leads to a strong influx of neutrophils in the vagina soon after the challenge which persisted despite the resolution of infection; ii) IL-17, produced by vaginal cells, particularly CD4 T cells, was detected in the vaginal wash during the infection, reaching a maximum 14 days after the challenge; iii) The amount and kinetics of IL-23 in vaginal fluids were comparable to those in vaginal cells; iv) The inhibition of Th17 differentiation led to significant inhibition of IL-17 production with consequent exacerbation of infection; v) An increased production of ?defensin 2 was manifested in cells of infected mice. This production was strongly reduced when Th17 differentiation was inhibited and was increased by rIL-17 treatment. Conclusions These results imply that IL-17 and Th17, along with innate antimicrobial factors, have a role in the immune response to vaginal candidiasis.

Pietrella, Donatella; Rachini, Anna; Pines, Mark; Pandey, Neelam; Mosci, Paolo; Bistoni, Francesco; d'Enfert, Cristophe; Vecchiarelli, Anna

2011-01-01

83

Epithelial cells remove apoptotic epithelial cells during post-lactation involution of the mouse mammary gland.  

PubMed

Following the cessation of lactation, the mammary gland undergoes a physiologic process of tissue remodeling called involution in which glandular structures are lost, leaving an adipose tissue compartment that takes up a much larger proportion of the tissue. A quantitative morphometric analysis was undertaken to determine the mechanisms for clearance of the epithelial cells during this process. The involution process was set in motion by removal of pups from 14-day lactating C57BL/6 mice. Within hours, milk-secreting epithelial cells were shed into the glandular lumen. These cells became apoptotic, exhibiting exposure of phosphatidylserine residues on their surfaces, activation of effector caspase-3, staining for caspase-cleaved keratin 18, loss of internal organellar structure, and nuclear breakdown, but minimal blebbing or generation of apoptotic bodies. Clearance of residual milk and the shed epithelial cells was rapid, with most of the removal occurring in the first 72 h. Intact apoptotic epithelial cells were engulfed in large numbers by residual viable epithelial cells into spacious efferosomes. This process led to essentially complete involution within 4 days, at which point estrous cycling recommenced. Macrophages and other inflammatory cells did not contribute to the clearance of either residual milk or apoptotic cells, which appeared to be due entirely to the epithelium itself. PMID:18057312

Monks, Jenifer; Smith-Steinhart, Christine; Kruk, Ellen R; Fadok, Valerie A; Henson, Peter M

2007-12-05

84

Epithelial cell proliferation in childhood enteropathies.  

PubMed Central

BACKGROUND/AIM: The aim of this study was to investigate epithelial cell turnover in childhood enteropathy to establish whether common disease related mechanisms operate. Levels of epithelial cell proliferation were measured in children with food intolerance (cows' milk protein intolerance and coeliac disease), and after infection with Giardia lamblia, Cryptosporidium, and enteropathogenic Escherichia coli. METHODS: Comparative measures of epithelial cell proliferation were performed by recording mitotic activity and MIB-1 immunoreactivity in proximal small intestinal biopsy specimens. RESULTS/CONCLUSIONS: A hyperplastic crypt response was evident in all of the disease states examined and was particularly pronounced in coeliac disease and in infection with enteropathogenic E coli, where mitotic and MIB-1 labelling indices were significantly raised above control values. In contrast with coeliac disease, increased crypt cell production rates in enteropathogenic E coli infection were also due to an expansion of the crypt proliferation compartment, without a comparable increase in crypt cell numbers. Crypt hyperplasia is therefore a common tissue response to mucosal damage in food allergy and infection, although disease specific mechanisms are evident. Images Figure 1: A-D Figure 1: E-F

Savidge, T C; Shmakov, A N; Walker-Smith, J A; Phillips, A D

1996-01-01

85

Force mapping in epithelial cell migration  

NASA Astrophysics Data System (ADS)

We measure dynamic traction forces exerted by epithelial cells on a substrate. The force sensor is a high-density array of elastomeric microfabricated pillars that support the cells. Traction forces induced by cell migration are deduced from the measurement of the bending of these pillars and are correlated with actin localization by fluorescence microscopy. We use a multiple-particle tracking method to estimate the mechanical activity of cells in real time with a high-spatial resolution (down to 2 µm) imposed by the periodicity of the post array. For these experiments, we use differentiated Madin-Darby canine kidney (MDCK) epithelial cells. Our data provide definite information on mechanical forces exerted by a cellular assembly. The maximum intensity of the forces is localized on the edge of the epithelia. Hepatocyte growth factor promotes cell motility and induces strong scattering activity of MDCK cells. Thus, we compare forces generated by MDCK cells in subconfluent epithelia versus isolated cells after hepatocyte growth factor treatment. Maximal-traction stresses at the edge of a monolayer correspond to higher values than those measured for a single cell and may be due to a collective behavior. cell mechanics | microfabrication | traction forces | multiple particle tracking

Du Roure, Olivia; Saez, Alexandre; Buguin, Axel; Austin, Robert H.; Chavrier, Philippe; Siberzan, Pascal; Ladoux, Benoit

2005-02-01

86

Gelatinase Secretion by Glomerular Epithelial Cells  

Microsoft Academic Search

A significant increase in gelatinolytic activity was observed in cultures of glomeruli from Heymann nephritic rats. Zymography of the culture medium indicated that the main gelatinase species has a molecular weight of 98 kilodaltons (kDa) and the characteristics of metalloproteinase. The 98-kDa gelatinase was detected in the culture medium of glomerular epithelial cells but not in those of endothelial and

Kazuyoshi Watanabe; Shigemi Kinoshita; Hideo Nakagawa

1990-01-01

87

12/15-lipoxygenase expressed in non-epithelial cells causes airway epithelial injury in asthma  

PubMed Central

The mechanisms underlying asthmatic airway epithelial injury are not clear. 12/15-lipoxygenase (an ortholog of human 15-LOX-1), which is induced by IL-13, is associated with mitochondrial degradation in reticulocytes at physiological conditions. In this study, we showed that 12/15-LOX expressed in nonepithelial cells caused epithelial injury in asthma pathogenesis. While 12/15-LOX overexpression or IL-13 administration to naïve mice showed airway epithelial injury, 12/15-LOX knockout/knockdown in allergic mice reduced airway epithelial injury. The constitutive expression of 15-LOX-1 in bronchial epithelia of normal human lungs further indicated that epithelial 15-LOX-1 may not cause epithelial injury. 12/15-LOX expression is increased in various inflammatory cells in allergic mice. Though non-epithelial cells such as macrophages or fibroblasts released 12/15-LOX metabolites upon IL-13 induction, bronchial epithelia didn't release. Further 12-S-HETE, arachidonic acid metabolite of 12/15-LOX leads to epithelial injury. These findings suggested 12/15-LOX expressed in non-epithelial cells such as macrophages and fibroblasts leads to bronchial epithelial injury.

Mabalirajan, Ulaganathan; Rehman, Rakhshinda; Ahmad, Tanveer; Kumar, Sarvesh; Leishangthem, Geeta Devi; Singh, Suchita; Dinda, Amit Kumar; Biswal, Shyam; Agrawal, Anurag; Ghosh, Balaram

2013-01-01

88

Characterization of mouse alveolar epithelial cell monolayers  

PubMed Central

We investigated the influence of extracellular matrix on transport properties of mouse alveolar epithelial cell (AEC) monolayers (MAECM) and transdifferentiation of isolated mouse alveolar epithelial type II (AT2) cells into an alveolar epithelial type I (AT1) cell-like phenotype. Primary mouse AT2 cells plated on laminin 5-coated polycarbonate filters formed monolayers with transepithelial resistance (RT) and equivalent short-circuit current (IEQ) of 1.8 k?·cm2 and 5.3 ?A/cm2, respectively, after 8 days in culture. Amiloride (10 ?M), ouabain (0.1 mM), and pimozide (10 ?M) decreased MAECM IEQ to 40%, 10%, and 65% of its initial value, respectively. Sequential addition of pimozide and amiloride, in either order, revealed that their inhibitory effects are additive, suggesting that cyclic nucleotide-gated channels contribute to amiloride-insensitive active ion transport across MAECM. Ussing chamber measurements of unidirectional ion fluxes across MAECM under short-circuit conditions indicated that net absorption of Na+ in the apical-to-basolateral direction is comparable to net ion flux calculated from the observed short-circuit current: 0.38 and 0.33 ?eq·cm?2·h?1, respectively. Between days 1 and 9 in culture, AEC demonstrated increased expression of aquaporin-5 protein, an AT1 cell marker, and decreased expression of pro-surfactant protein-C protein, an AT2 cell marker, consistent with transition to an AT1 cell-like phenotype. These results demonstrate that AT1 cell-like MAECM grown on laminin 5-coated polycarbonate filters exhibit active and passive transport properties that likely reflect the properties of intact mouse alveolar epithelium. This mouse in vitro model will enhance the study of AEC derived from mutant strains of mice and help define important structure-function correlations.

DeMaio, Lucas; Tseng, Wanru; Balverde, Zerlinde; Alvarez, Juan R.; Kim, Kwang-Jin; Kelley, Diane G.; Senior, Robert M.; Crandall, Edward D.; Borok, Zea

2009-01-01

89

Separation of sperm and vaginal cells with flow cytometry for DNA typing after sexual assault  

Microsoft Academic Search

Background: Successful DNA typing after rape is limited when only a few sperm and numerous vaginal cells are recovered from a swab, resulting in an extremely unfavorable ratio of male to female DNA. The goal of this study was to develop a protocol involving sperm cell sorting with flow cytometry based on differences in ploidy, major histocompatibility class I, CD45,

Wolfgang M. J Schoell; Michael Klintschar; Ramin Mirhashemi; Barbara Pertl

1999-01-01

90

Epithelial BMP signaling is required for proper specification of epithelial cell lineages and gastric endocrine cells.  

PubMed

Bone morphogenetic protein (BMP) signaling within the gastrointestinal tract is complex. BMP ligands and their receptors are expressed in both epithelial and mesenchymal compartments, suggesting bidirectional signaling between these two entities. Despite an increasing interest in BMP signaling in gut physiology and pathologies, the distinct contribution of BMP signaling in the epithelium vs. the mesenchyme in gastrointestinal homeostasis remains to be established. We aimed to investigate the role of epithelial BMP signaling in gastric organogenesis, gland morphogenesis, and maintenance of epithelial cell functions. Using the Cre/loxP system, we generated a mouse model with an early deletion during development of BMP receptor 1A (Bmpr1a) exclusively in the foregut endoderm. Bmpr1a(?GEC) mice showed no severe abnormalities in gastric organogenesis, gland epithelial proliferation, or morphogenesis, suggesting only a minor role for epithelial BMP signaling in these processes. However, early loss of BMP signaling in foregut endoderm did impact on gastric patterning, leading to an anteriorization of the stomach. In addition, numbers of parietal cells were reduced in Bmpr1a(?GEC) mice. Epithelial BMP deletion significantly increased the numbers of chromogranin A-, ghrelin-, somatostatin-, gastrin-, and serotonin-expressing gastric endocrine cells. Cancer never developed in young adult (<100 days) Bmpr1a-inactivated mice although a marker of spasmolytic polypeptide-expressing metaplasia was upregulated. Using this model, we have uncovered that BMP signaling negatively regulates the proliferation and commitment of endocrine precursor cells. Our data also indicate that loss of BMP signaling in epithelial gastric cells alone is not sufficient to induce gastric neoplasia. PMID:21415412

Maloum, Faïza; Allaire, Joannie M; Gagné-Sansfaçon, Jessica; Roy, Evelyne; Belleville, Karine; Sarret, Philippe; Morisset, Jean; Carrier, Julie C; Mishina, Yuji; Kaestner, Klaus H; Perreault, Nathalie

2011-03-17

91

Epithelial BMP signaling is required for proper specification of epithelial cell lineages and gastric endocrine cells  

PubMed Central

Bone morphogenetic protein (BMP) signaling within the gastrointestinal tract is complex. BMP ligands and their receptors are expressed in both epithelial and mesenchymal compartments, suggesting bidirectional signaling between these two entities. Despite an increasing interest in BMP signaling in gut physiology and pathologies, the distinct contribution of BMP signaling in the epithelium vs. the mesenchyme in gastrointestinal homeostasis remains to be established. We aimed to investigate the role of epithelial BMP signaling in gastric organogenesis, gland morphogenesis, and maintenance of epithelial cell functions. Using the Cre/loxP system, we generated a mouse model with an early deletion during development of BMP receptor 1A (Bmpr1a) exclusively in the foregut endoderm. Bmpr1a?GEC mice showed no severe abnormalities in gastric organogenesis, gland epithelial proliferation, or morphogenesis, suggesting only a minor role for epithelial BMP signaling in these processes. However, early loss of BMP signaling in foregut endoderm did impact on gastric patterning, leading to an anteriorization of the stomach. In addition, numbers of parietal cells were reduced in Bmpr1a?GEC mice. Epithelial BMP deletion significantly increased the numbers of chromogranin A-, ghrelin-, somatostatin-, gastrin-, and serotonin-expressing gastric endocrine cells. Cancer never developed in young adult (<100 days) Bmpr1a-inactivated mice although a marker of spasmolytic polypeptide-expressing metaplasia was upregulated. Using this model, we have uncovered that BMP signaling negatively regulates the proliferation and commitment of endocrine precursor cells. Our data also indicate that loss of BMP signaling in epithelial gastric cells alone is not sufficient to induce gastric neoplasia.

Maloum, Faiza; Allaire, Joannie M.; Gagne-Sansfacon, Jessica; Roy, Evelyne; Belleville, Karine; Sarret, Philippe; Morisset, Jean; Carrier, Julie C.; Mishina, Yuji; Kaestner, Klaus H.

2011-01-01

92

Vaginal itching  

MedlinePLUS

... and make you more susceptible to infections. Vaginal yeast infection Vaginitis . Vaginitis in girls before puberty is ... If you are sure that you have a yeast infection, try over-the-counter creams or vaginal ...

93

Analysis of epithelial barrier integrity in polarized lung epithelial cells.  

PubMed

Epithelial surfaces of the body are a key component of host defense by providing a mechanical barrier against potentially harmful substances. The respiratory tract is constantly challenged by a wide range of airborne pathogens and particulates, and provides not only a mucosal barrier, but also an intricate innate immune defense system. Disruption of the alveolar epithelial barrier can lead to acute lung injury, pneumonia, and acute respiratory distress syndrome. Protection of lung epithelial integrity, or repair of hyperpermeability with keratinocyte growth factor or Hsp90 inhibitors, is crucial for combating permeability edema. Ex vivo-differentiated lung epithelium represents a physiologically relevant tool for analyzing the effect of pathogens, chemicals, or drugs on lung barrier function. The integrity of the lung epithelial layer can be determined by several approaches. By combining two of these techniques, transepithelial electrical resistance and paracellular flux of fluorescent molecules, information about barrier integrity can be obtained in a prompt and convenient manner. As example, the virus- or bacterial toxin-mediated disruption of an ex vivo-differentiated mucociliary lung epithelial barrier is used here for assessing advantages and limitations of these methods. PMID:21874453

Strengert, Monika; Knaus, Ulla G

2011-01-01

94

Products for Human and Mouse Mammary Epithelial Cell Research  

Microsoft Academic Search

+ CD10 - ). These distinct subpopulations can be isolated from normal mammary tissue or from mammary epithelial cell cultures using either flow cytometry or immunomagnetic cell separation. The growth and differentiation potential of these cells can be monitored by their ability to form colonies when plated at low density in vitro. Mammary Epithelial Cell Enrichment and Culture StemCell Technologies

Carmine Alum

2006-01-01

95

Interferons Mediate Terminal Differentiation of Human Cortical Thymic Epithelial Cells  

Microsoft Academic Search

In the thymus, epithelial cells comprise a heterogeneous population required for the generation of functional T lymphocytes, suggesting that thymic epithelium disruption by viruses may compromise T-cell lymphopoiesis in this organ. In a previous report, we demonstrated that in vitro, measles virus induced differentiation of cortical thymic epithelial cells as characterized by (i) cell growth arrest, (ii) morphological and phenotypic

Pierre-Olivier Vidalain; David Laine; Yona Zaffran; Olga Azocar; Christine Servet-Delprat; T. Fabian Wild; Chantal Rabourdin-Combe; Hélène Valentin

2002-01-01

96

Pathways of differentiation of airway epithelial cells.  

PubMed Central

The question being examined is whether one or more morphologically distinct cell types can be identified in the conducting airways of adult rabbits possessing stem cell functions. The term "stem cell" is used to denote cells with extensive self-replicating potential and the ability to produce differentiated progeny. According to various models of cell renewal in the conducting airways that have been proposed over the years, two different cell types have to be regarded as primary candidates for the stem cell: basal cells and some type of secretory cells. The question is complicated by the fact that significant differences exist between species in the distribution and morphological characteristics of airway cell types. In addition, different airway segments may or may not be occupied by different populations of stem cells. Previously, investigators have addressed the problem by studying normal cell regeneration or injury induced cell regeneration in vivo in the whole animal. We decided to attempt a different approach, namely, to separate specific cell types and to study the proliferation and differentiation capacity of such cell isolates using in vitro and in vivo cell culture techniques. Our studies lead us to conclude that the conducting airways of adult rabbits contain at least two distinct cell populations endowed with stem cell potential, namely basal cells and bronchiolar Clara cells. From that it follows that the trachea and bronchi, on one hand, and the bronchioles, on the other hand, are occupied by two different stem cell populations governing renewal of the epithelial lining. Images PLATE 1. PLATE 2. PLATE 3. A PLATE 3. B PLATE 4. PLATE 5. PLATE 6. A PLATE 6. B PLATE 7. PLATE 8. PLATE 9. A PLATE 9. B PLATE 10. A PLATE 10. B PLATE 11. A PLATE 11. B PLATE 11. C PLATE 11. D

Nettesheim, P; Jetten, A M; Inayama, Y; Brody, A R; George, M A; Gilmore, L B; Gray, T; Hook, G E

1990-01-01

97

Spontaneous Production of Immunoglobulin M in Human Epithelial Cancer Cells  

PubMed Central

It is well known that B-1 B cells are the main cell type that is responsible for the production of natural immunoglobulin M (IgM) and can respond to infection by increasing IgM secretion. However, we unexpectedly found that some epithelial cells also can express rearranged IgM transcript that has natural IgM characteristics, such as germline-encoded and restricted rearrangement patterns. Here we studied IgM expression in human non-B cells and found that IgM was frequently expressed by many human epithelial cancer cells as well as non-cancer epithelial cells. Moreover, CD79A and CD79B, two molecules that are physically linked to membranous IgM on the surface of B cells to form the B cell antigen receptor complex, were also expressed on the cell surface of epithelial cancer cells and co-located with IgM. Like the natural IgM, the epithelial cancer cell-derived IgM recognized a series of microbial antigens, such as single-stranded DNA, double-stranded DNA, lipopolysaccharide, and the HEp-2 cell antigen. More important, stimulation of the toll-like receptor 9 (TLR9), which mimics bacterial infection, substantially increased the secretion of IgM in human epithelial cancer cells. These findings indicate that human epithelial cancer cells as well as non-cancer epithelial cells can spontaneously produce IgM with natural antibody activity.

Hu, Fanlei; Zhang, Li; Zheng, Jie; Zhao, Ling; Huang, Jing; Shao, Wenwei; Liao, Qinyuan; Ma, Teng; Geng, Li; Yin, C. Cameron; Qiu, Xiaoyan

2012-01-01

98

Transport of IGF-I across epithelial cell monolayers  

Microsoft Academic Search

Epithelial cells line the lumens of organs including the gastrointestinal tract, kidney tubules and respiratory airways, where they regulate the transport of electrolytes and the movement of macromolecules. The current study aimed to investigate the transport of IGF-I across epithelial cell barriers. Epithelial cell lines derived from gut (IEC-6), kidney (MDBK) and lung (Mv1Lu) were shown to possess high-aYnity, functional

S E P Bastian; P E Walton; F J Ballard; D A Belford

1999-01-01

99

Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties  

Microsoft Academic Search

The epithelial compartment of the human breast comprises two distinct lineages: the luminal epithelial and the myoepithelial lineage. We have shown previously that a subset of the luminal epithelial cells could convert to myoepithelial cells in culture signifying the possible existence of a progenitor cell. We therefore set out to identify and isolate the putative precursor in the luminal epithelial

Thorarinn Gudjonsson; Helga Lind Nielsen; Lone Rønnov-Jessen; Mina J. Bissell; Ole William Petersen

2002-01-01

100

Ouabain modulates ciliogenesis in epithelial cells  

PubMed Central

The exchange of substances between higher organisms and the environment occurs across transporting epithelia whose basic features are tight junctions (TJs) that seal the intercellular space, and polarity, which enables cells to transport substances vectorially. In a previous study, we demonstrated that 10 nM ouabain modulates TJs, and we now show that it controls polarity as well. We gauge polarity through the development of a cilium at the apical domain of Madin-Darby canine kidney cells (MDCK, epithelial dog kidney). Ouabain accelerates ciliogenesis in an ERK1/2-dependent manner. Claudin-2, a molecule responsible for the Na+ and H2O permeability of the TJs, is also present at the cilium, as it colocalizes and coprecipitates with acetylated ?-tubulin. Ouabain modulates claudin-2 localization at the cilium through ERK1/2. Comparing wild-type and ouabain-resistant MDCK cells, we show that ouabain acts through Na+,K+-ATPase. Taken together, our previous and present results support the possibility that ouabain constitutes a hormone that modulates the transporting epithelial phenotype, thereby playing a crucial role in metazoan life.

Larre, Isabel; Castillo, Aida; Flores-Maldonado, Catalina; Contreras, Ruben G.; Galvan, Ivan; Munoz-Estrada, Jesus; Cereijido, Marcelino

2011-01-01

101

Current perspectives in epithelial cell injury and repair: consequences for epithelial  

Microsoft Academic Search

Epithelial cells lining the airways and the respiratory compartment may, and certainly when exposed to an inflammatory milieu, display an altered functioning, which could contribute to pathophysiology of inflammatory lung\\/airway disease. In the present review paper, several issues that were discussed at an earlier European Respiratory Society Research Seminar on conditions that affect epithelial functioning have been recapitulated and updated.

R. Lutter; M. Spiteri

102

Stromal-epithelial interactions in aging and cancer: Senescent fibroblasts alter epithelial cell differentiation  

SciTech Connect

Cellular senescence suppresses cancer by arresting cells at risk for malignant tumorigenesis. However, senescent cells also secrete molecules that can stimulate premalignant cells to proliferate and form tumors, suggesting the senescence response is antagonistically pleiotropic. We show that premalignant mammary epithelial cells exposed to senescent human fibroblasts in mice irreversibly lose differentiated properties, become invasive and undergo full malignant transformation. Moreover, using cultured mouse or human fibroblasts and non-malignant breast epithelial cells, we show that senescent fibroblasts disrupt epithelial alveolar morphogenesis, functional differentiation, and branching morphogenesis. Further, we identify MMP-3 as the major factor responsible for the effects of senescent fibroblasts on branching morphogenesis. Our findings support the idea that senescent cells contribute to age-related pathology, including cancer, and describe a new property of senescent fibroblasts--the ability to alter epithelial differentiation--that might also explain the loss of tissue function and organization that is a hallmark of aging.

Parrinello, Simona; Coppe, Jean-Philippe; Krtolica, Ana; Campisi, Judith

2004-07-14

103

CsrRS and environmental pH regulate group B streptococcus adherence to human epithelial cells and extracellular matrix.  

PubMed

Streptococcus agalactiae (group B Streptococcus or GBS) is a common colonizer of the gastrointestinal and genital tracts and an important cause of invasive infections in newborn infants and in adults with predisposing chronic conditions or advanced age. Attachment to epithelial surfaces at mucosal sites is a critical step in the successful colonization of a human host, and regulation of this process is likely to play an important role in both commensalism and dissemination to cause invasive disease. We found that inactivation of the CsrRS (or CovRS) two-component system increased GBS adherence to epithelial cells derived from human vaginal, cervical, and respiratory epithelium, as well as increasing adherence to extracellular matrix proteins and increasing biofilm formation on polystyrene. Neutral (as opposed to acidic) pH enhanced GBS binding to vaginal epithelial cells and to fibrinogen and fibronectin, effects that were partially dependent on CsrRS. The regulatory effects of CsrRS and environmental pH on bacterial adherence correlated with their effects on the expression of multiple surface adhesins, as assessed by quantitative reverse transcription-PCR. We conclude that GBS adherence to epithelial and abiotic surfaces is regulated by the CsrRS two-component system and by environmental pH through their regulatory effects on the expression of bacterial surface adhesins. Dynamic regulation of GBS adherence enhances the organism's adaptability to survival in multiple niches in the human host. PMID:22949550

Park, Su Eun; Jiang, Shengmei; Wessels, Michael R

2012-09-04

104

Mapping of HNF4? target genes in intestinal epithelial cells  

Microsoft Academic Search

BACKGROUND: The role of HNF4? has been extensively studied in hepatocytes and pancreatic ?-cells, and HNF4? is also regarded as a key regulator of intestinal epithelial cell differentiation. The aim of the present work is to identify novel HNF4? target genes in the human intestinal epithelial cells in order to elucidate the role of HNF4? in the intestinal differentiation progress.

Mette Boyd; Simon Bressendorff; Jette Møller; Jørgen Olsen; Jesper T Troelsen

2009-01-01

105

Identification and Characterization of Thymic Epithelial Progenitor Cells  

Microsoft Academic Search

T cell differentiation and repertoire selection depend critically on several distinct thymic epithelial cell types, whose lineage relationships are unclear. We have investigated these relationships via functional analysis of the epithelial populations within the thymic primordium. Here, we show that mAbs MTS20 and MTS24 identify a population of cells that, when purified and grafted ectopically, can differentiate into all known

Andrea R. Bennett; Alison Farley; Natalie F. Blair; Julie Gordon; Linda Sharp; C. Clare Blackburn

2002-01-01

106

Henipavirus pathogenesis in human respiratory epithelial cells.  

PubMed

Hendra virus (HeV) and Nipah virus (NiV) are deadly zoonotic viruses for which no vaccines or therapeutics are licensed for human use. Henipavirus infection causes severe respiratory illness and encephalitis. Although the exact route of transmission in human is unknown, epidemiological studies and in vivo studies suggest that the respiratory tract is important for virus replication. However, the target cells in the respiratory tract are unknown, as are the mechanisms by which henipaviruses can cause disease. In this study, we characterized henipavirus pathogenesis using primary cells derived from the human respiratory tract. The growth kinetics of NiV-Malaysia, NiV-Bangladesh, and HeV were determined in bronchial/tracheal epithelial cells (NHBE) and small airway epithelial cells (SAEC). In addition, host responses to infection were assessed by gene expression analysis and immunoassays. Viruses replicated efficiently in both cell types and induced large syncytia. The host response to henipavirus infection in NHBE and SAEC highlighted a difference in the inflammatory response between HeV and NiV strains as well as intrinsic differences in the ability to mount an inflammatory response between NHBE and SAEC. These responses were highest during HeV infection in SAEC, as characterized by the levels of key cytokines (interleukin 6 [IL-6], IL-8, IL-1?, monocyte chemoattractant protein 1 [MCP-1], and colony-stimulating factors) responsible for immune cell recruitment. Finally, we identified virus strain-dependent variability in type I interferon antagonism in NHBE and SAEC: NiV-Malaysia counteracted this pathway more efficiently than NiV-Bangladesh and HeV. These results provide crucial new information in the understanding of henipavirus pathogenesis in the human respiratory tract at an early stage of infection. PMID:23302882

Escaffre, Olivier; Borisevich, Viktoriya; Carmical, J Russ; Prusak, Deborah; Prescott, Joseph; Feldmann, Heinz; Rockx, Barry

2013-01-09

107

Epithelial Cell Secretions from the Human Female Reproductive Tract Inhibit Sexually Transmitted Pathogens and Candida albicans but not Lactobacillus  

PubMed Central

Female reproductive tract (FRT) epithelial cells protect against potential pathogens and sexually transmitted infections. The purpose of this study was to determine if epithelial cells from the upper FRT secrete antimicrobials that inhibit reproductive tract pathogens which threaten women's health. Apical secretions from primary cultures of Fallopian tube, uterine, cervical and ectocervical epithelial cells were incubated with Neisseria gonorrhoeae, Candida albicans (yeast and hyphal forms), HIV-1, and Lactobacillus crispatus, prior to being tested for their ability to grow and/or infect target cells. Epithelial cell secretions from the upper FRT inhibit N. gonorrhoeae and both forms of Candida, as well as reduce HIV-1 (R5) infection of target cells. In contrast, none had an inhibitory effect on L. crispatus. Cytokines and chemokines analysis in uterine secretions revealed several molecules that could account for pathogen inhibition. These findings provide definitive evidence for the critical role of epithelial cells in protecting the FRT from infections, without comprising the beneficial presence of L. crispatus, which is part of the normal vaginal microflora of humans.

Wira, CR; Ghosh, M; Smith, JM; Shen, L; Connor, RI; Sundstrom, P; Frechette, Gregory M.; Hill, EM; Fahey, JV

2011-01-01

108

Epithelial cell polarity: a major gatekeeper against cancer?  

PubMed Central

The correct establishment and maintenance of cell polarity are crucial for normal cell physiology and tissue homeostasis. Conversely, loss of cell polarity, tissue disorganisation and excessive cell growth are hallmarks of cancer. In this review, we focus on identifying the stages of tumoural development that are affected by the loss or deregulation of epithelial cell polarity. Asymmetric division has recently emerged as a major regulatory mechanism that controls stem cell numbers and differentiation. Links between cell polarity and asymmetric cell division in the context of cancer will be examined. Apical–basal polarity and cell–cell adhesion are tightly interconnected. Hence, how loss of cell polarity in epithelial cells may promote epithelial mesenchymal transition and metastasis will also be discussed. Altogether, we present the argument that loss of epithelial cell polarity may have an important role in both the initiation of tumourigenesis and in later stages of tumour development, favouring the progression of tumours from benign to malignancy.

Royer, C; Lu, X

2011-01-01

109

Nuclear microscopy of rat colon epithelial cells  

NASA Astrophysics Data System (ADS)

Using Nuclear microscopy, we have investigated iron distributions in the colons of Sprague Dawley rats, in order to elucidate heme uptake. Four groups of five Sprague Dawley rats (mean weight 180 g) were fed different purified diets containing either heme diet (2.5% w/w hemoglobin), high fat diet (HFD) (18% w/w fat, 1% w/w cholesterol), 'western' diet (combination of hemoglobin 2.5% and 18% fat, 1% cholesterol) or control diet (7% w/w fat). After 4 weeks, animals were sacrificed by exsanguination after anaesthesia.Thin sections of frozen colon tissue were taken, freeze dried and scanned using nuclear microscopy utilising the techniques PIXE, RBS and STIM. The new data acquisition system (IonDaq) developed in CIBA was used to obtain high resolution images and line scans were used to map the iron distributions across the colon boundaries.The nuclear microscope results indicate that when HFD is given in addition to heme, the iron content of the epithelial cells that line the colon decreases, and the zinc in the smooth muscle wall increases. This implies that the level of heme and fat in diet has an important role in colon health, possibly by influencing epithelial cells directly or changing luminal composition such as bacterial flora or levels of metabolites and cytotoxins.

Ren, M.; Rajendran, Reshmi; Ng, Mary; Udalagama, Chammika; Rodrigues, Anna E.; Watt, Frank; Jenner, Andrew Michael

2011-10-01

110

Ouabain modulates epithelial cell tight junction  

PubMed Central

Epithelial cells treated with high concentrations of ouabain (e.g., 1 ?M) retrieve molecules involved in cell contacts from the plasma membrane and detach from one another and their substrates. On the basis of this observation, we suggested that ouabain might also modulate cell contacts at low, nontoxic levels (10 or 50 nM). To test this possibility, we analyzed its effect on a particular type of cell–cell contact: the tight junction (TJ). We demonstrate that at concentrations that neither inhibit K+ pumping nor disturb the K+ balance of the cell, ouabain modulates the degree of sealing of the TJ as measured by transepithelial electrical resistance (TER) and the flux of neutral 3 kDa dextran (JDEX). This modulation is accompanied by changes in the levels and distribution patterns of claudins 1, 2, and 4. Interestingly, changes in TER, JDEX, and claudins behavior are mediated through signal pathways containing ERK1/2 and c-Src, which have distinct effects on each physiological parameter and claudin type. These observations support the theory that at low concentrations, ouabain acts as a modulator of cell–cell contacts.

Larre, Isabel; Lazaro, Amparo; Contreras, Ruben G.; Balda, Maria S.; Matter, Karl; Flores-Maldonado, Catalina; Ponce, Arturo; Flores-Benitez, David; Rincon-Heredia, Ruth; Padilla-Benavides, Teresita; Castillo, Aida; Shoshani, Liora; Cereijido, Marcelino

2010-01-01

111

Anthrax Toxin Entry into Polarized Epithelial Cells  

PubMed Central

We examined the entry of anthrax edema toxin (EdTx) into polarized human T84 epithelial cells using cyclic AMP-regulated Cl? secretion as an index of toxin entry. EdTx is a binary A/B toxin which self assembles at the cell surface from anthrax edema factor and protective antigen (PA). PA binds to cell surface receptors and delivers EF, an adenylate cyclase, to the cytosol. EdTx elicited a strong Cl? secretory response when it was applied to the basolateral surface of T84 cells but no response when it was applied to the apical surface. PA alone had no effect when it was applied to either surface. T84 cells exposed basolaterally bound at least 30-fold-more PA than did T84 cells exposed apically, indicating that the PA receptor is largely or completely restricted to the basolateral membrane of these cells. The PA receptor did not fractionate with detergent-insoluble caveola-like membranes as cholera toxin receptors do. These findings have implications regarding the nature of the PA receptor and confirm the view that EdTx and CT coopt fundamentally different subcellular systems to enter the cell and cause disease.

Beauregard, Kathryn E.; Wimer-Mackin, Susan; Collier, R. John; Lencer, Wayne I.

1999-01-01

112

Epithelial-mesenchymal transition can suppress major attributes of human epithelial tumor-initiating cells.  

PubMed

Malignant progression in cancer requires populations of tumor-initiating cells (TICs) endowed with unlimited self renewal, survival under stress, and establishment of distant metastases. Additionally, the acquisition of invasive properties driven by epithelial-mesenchymal transition (EMT) is critical for the evolution of neoplastic cells into fully metastatic populations. Here, we characterize 2 human cellular models derived from prostate and bladder cancer cell lines to better understand the relationship between TIC and EMT programs in local invasiveness and distant metastasis. The model tumor subpopulations that expressed a strong epithelial gene program were enriched in highly metastatic TICs, while a second subpopulation with stable mesenchymal traits was impoverished in TICs. Constitutive overexpression of the transcription factor Snai1 in the epithelial/TIC-enriched populations engaged a mesenchymal gene program and suppressed their self renewal and metastatic phenotypes. Conversely, knockdown of EMT factors in the mesenchymal-like prostate cancer cell subpopulation caused a gain in epithelial features and properties of TICs. Both tumor cell subpopulations cooperated so that the nonmetastatic mesenchymal-like prostate cancer subpopulation enhanced the in vitro invasiveness of the metastatic epithelial subpopulation and, in vivo, promoted the escape of the latter from primary implantation sites and accelerated their metastatic colonization. Our models provide new insights into how dynamic interactions among epithelial, self-renewal, and mesenchymal gene programs determine the plasticity of epithelial TICs. PMID:22505459

Celià-Terrassa, Toni; Meca-Cortés, Oscar; Mateo, Francesca; de Paz, Alexia Martínez; Rubio, Nuria; Arnal-Estapé, Anna; Ell, Brian J; Bermudo, Raquel; Díaz, Alba; Guerra-Rebollo, Marta; Lozano, Juan José; Estarás, Conchi; Ulloa, Catalina; Álvarez-Simón, Daniel; Milà, Jordi; Vilella, Ramón; Paciucci, Rosanna; Martínez-Balbás, Marian; de Herreros, Antonio García; Gomis, Roger R; Kang, Yibin; Blanco, Jerónimo; Fernández, Pedro L; Thomson, Timothy M

2012-04-16

113

Goat uterine epithelial cells are susceptible to infection with Caprine Arthritis Encephalitis Virus (CAEV) in vivo  

PubMed Central

The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.

2012-01-01

114

The status of the surface enzyme guanidinobenzoatase on mature epithelial cells in sputum cell monolayers.  

PubMed

Mature epithelial cells derived from the sputum of normal healthy adults lack GB, while the epithelial cells of sputum collected from patients with lung tumours contain a spectrum of epithelial cells with active GB, some of which are clearly tumour cells, based upon their morphology. The presence of abnormal epithelial cells was confirmed by the cytologist whilst observing the fluorescent stained cells and later by independent cytological analysis of these same cells employing conventional stains. PMID:8517644

Steven, F S; Payne, P W; Palcic, B

115

Airway epithelial IL-15 transforms monocytes into dendritic cells.  

PubMed

IL-15 has recently been shown to induce the differentiation of functional dendritic cells (DCs) from human peripheral blood monocytes. Since DCs lay in close proximity to epithelial cells in the airway mucosa, we investigated whether airway epithelial cells release IL-15 in response to inflammatory stimuli and thereby induce differentiation and maturation of DCs. Alveolar (A549) and bronchial (BEAS-2B) epithelial cells produced IL-15 spontaneously and in a time- and dose-dependent manner after stimulation with IL-1beta, IFN-gamma, or TNF-alpha. Airway epithelial cell supernatants induced an increase of IL-15Ralpha gene expression in ex vivo monocytes, and stimulated DCs enhanced their IL-15Ralpha gene expression up to 300-fold. Airway epithelial cell-conditioned media induced the differentiation of ex vivo monocytes into partially mature DCs (HLA-DR+, DC-SIGN+, CD14+, CD80-, CD83+, CD86+, CCR3+, CCR6(+), CCR7-). Based on their phenotypic (CD123+, BDCA2+, BDCA4+, BDCA1(-), CD1a-) and functional properties (limited maturation upon stimulation with LPS and limited capacity to induce T cell proliferation), these DCs resembled plasmacytoid DCs. The effects of airway epithelial cell supernatants were largely blocked by a neutralizing monoclonal antibody to IL-15. Thus, our results demonstrate that airway epithelial cell-conditioned media have the capacity to differentiate monocytes into functional DCs, a process substantially mediated by epithelial-derived IL-15. PMID:17363780

Regamey, Nicolas; Obregon, Carolina; Ferrari-Lacraz, Sylvie; van Leer, Coretta; Chanson, Marc; Nicod, Laurent P; Geiser, Thomas

2007-03-15

116

Cervical mucins carry alpha(1,2)fucosylated glycans that partly protect from experimental vaginal candidiasis.  

PubMed

Cervical mucins are glycosylated proteins that form a protective cervical mucus. To understand the role of mucin glycans in Candida albicans infection, oligosaccharides from mouse cervical mucins were analyzed by liquid chromatography-mass spectrometry. Cervical mucins carry multiple alpha(1-2)fucosylated glycans, but alpha(1,2)fucosyltransferase Fut2-null mice are devoid of these epitopes. Epithelial cells in vaginal lavages from Fut2-null mice lacked Ulex europaeus agglutinin-1 (UEA-I) staining for alpha(1-2)fucosylated glycans. Hysterectomy to remove cervical mucus eliminated UEA-I and acid mucin staining in vaginal epithelial cells from wild type mice indicating the cervix as the source of UEA-I positive epithelial cells. To assess binding of alpha(1-2) fucosylated glycans on C. albicans infection, an in vitro adhesion assay was performed with vaginal epithelial cells from wild type and Fut2-null mice. Vaginal epithelial cells from Fut2-null mice were found to bind increased numbers of C. albicans compared to vaginal epithelial cells obtained from wild type mice. Hysterectomy lessened the difference between Fut2-null and wild type mice in binding of C. ablicans in vitro and susceptibility to experimental C. albicans vaginitis in vivo. We generated a recombinant fucosylated MUC1 glycanpolymer to test whether the relative protection of wild type mice compared to Fut2-null mice could be mimicked with exogenous mucin. While a small portion of the recombinant MUC1 epitopes displayed alpha(1-2)fucosylated glycans, the predominant epitopes were sialylated due to endogenous sialyltransferases in the cultured cells. Intravaginal instillation of recombinant MUC1 glycanpolymer partially reduced experimental yeast vaginitis suggesting that a large glycanpolymer, with different glycan epitopes, may affect fungal burden. PMID:19326211

Domino, Steven E; Hurd, Elizabeth A; Thomsson, Kristina A; Karnak, David M; Holmén Larsson, Jessica M; Thomsson, Elisabeth; Bäckström, Malin; Hansson, Gunnar C

2009-12-01

117

Intracellular degradation of Fusobacterium nucleatum in human gingival epithelial cells  

Microsoft Academic Search

The role of Fusobacterium nucleatum in oral health and disease is controversial. We have previously shown that F. nucleatum invades gingival epithelial cells. However, the destiny of the internalized F. nucleatum is not clear. In the present study, the intracellular destiny of F. nucleatum and its cytopathic effect on gingival epithelial cells were studied. The ability of F. nucleatum and

Suk Ji; Ji Eun Shin; Yong Cheol Kim; Youngnim Choi

2010-01-01

118

Development and characterization of rabbit proximal tubular epithelial cell lines  

Microsoft Academic Search

Development and characterization of rabbit proximal tubular epithelial cell lines. We have isolated rabbit kidney proximal tubular epithelial cell lines. The selection was based on their ability to form confluent monolayers on porous supports and to maintain receptor-mediated signal transduction and ion transport, characteristic of the proximal tubule. The isolation method consisted of several steps: (1) superficial cortical proximal tubule

Michael F Romero; Janice G Douglas; Richard L Eckert; Ulrich Hopfer; James W Jacobberger

1992-01-01

119

Rhinovirus Disrupts the Barrier Function of Polarized Airway Epithelial Cells  

Microsoft Academic Search

Rationale : Secondary bacterial infection following rhinovirus (RV) in- fection has been recognized in chronic obstructive pulmonary disease. Objectives: We sought to understand mechanisms by which RV in- fection facilitates secondary bacterial infection. Methods: Primary human airway epithelial cells grown at air-liquid interface and human bronchial epithelial (16HBE14o-) cells grown as polarized monolayers were infected apically with RV. Transmigration of

Umadevi Sajjan; Qiong Wang; Ying Zhao; Dieter C. Gruenert; Marc B. Hershenson

120

Effect of Rabbit Epididymal Antimicrobial Peptide, REHb?P, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal Cells In Vitro  

PubMed Central

Antimicrobial peptides (AMP's) protect epithelial surfaces including epididymis against pathogens and play a key role in orchestrating various defensive responses. Recently, we have identified one such AMP, rabbit epididymal hemoglobin-? subuit (REHb?P) from the epididymal fluid of rabbit, Oryctologus cuniculus. The demonstration of a protective role of REHb?P in epididymal epithelial cells (EPEC's) led us to investigate: (1) the identification of LPS interactive domain in REHb?P, and (2) whether the REHb?P of rabbit origin mediates vaginal cellular immune responses of another species (human). HeLa-S3, human vaginal epithelial cells (hVECs) were exposed to LPS or the LPS-stimulated cells treated with REHb?P or neutral peptide, nREHb?P. Effect of LPS and cytokines (IL-6 and IL-1?) and chemokines (IL-8, MCP-1) levels was determined in the culture supernatants. In response to the LPS, hVECs synthesized these mediators and the levels were significantly higher than controls. This enhancing effect was ameliorated when the LPS-induced hVECs were treated with REHb?P. Similar results were obtained on NF-?B protein and hBD-1 mRNA expression. Confocal microscopy studies revealed that REHb?P attenuated the LPS-induced internalization of E. coli by macrophages. The chemotaxis studies performed using Boyden chamber Transwell assay, which showed elevated migration of U937 cells when the supernatants of LPS-induced hVECs were used, and the effect was inhibited by REHb?P. REHb?P was found to be localized on the acrosome of rabbit spermatozoa, suggesting its role in sperm protection beside sperm function. In conclusion, REHb?P may have the potential to develop as a therapeutic agent for reproductive tract infections (RTI's).

Reddy, K. V. R.; Sukanya, D.; Patgaonkar, M. S.; Selvaakumar, C.

2012-01-01

121

Sodium selectivity of Reissner's membrane epithelial cells  

PubMed Central

Background Sodium absorption by Reissner's membrane is thought to contribute to the homeostasis of the volume of cochlear endolymph. It was previously shown that the absorptive transepithelial current was blocked by amiloride and benzamil. The most commonly-observed target of these drugs is the epithelial sodium channel (ENaC), which is composed of the three subunits ?-,?- and ?-ENaC. However, other less-selective cation channels have also been observed to be sensitive to benzamil and amiloride. The aim of this study was to determine whether Reissner's membrane epithelial cells could support parasensory K+ absorption via amiloride- and benzamil-sensitive electrogenic pathways. Results We determined the molecular and functional expression of candidate cation channels with gene array (GEO GSE6196), RT-PCR, and whole-cell patch clamp. Transcript expression analysis of Reissner's membrane detected no amiloride-sensitive acid-sensing ion channels (ASIC1a, ASIC2a, ASIC2b) nor amiloride-sensitive cyclic-nucleotide gated channels (CNGA1, CNGA2, CNGA4, CNGB3). By contrast, ?-,?- and ?-ENaC were all previously reported as present in Reissner's membrane. The selectivity of the benzamil-sensitive cation currents was observed in whole-cell patch clamp recordings under Cl--free conditions where cations were the only permeant species. The currents were carried by Na+ but not K+, and the permeability of Li+ was greater than that of Na+ in Reissner's membrane. Complete replacement of bath Na+ with the inpermeable cation NMDG+ led to the same inward current as with benzamil in a Na+ bath. Conclusions These results are consistent with the amiloride/benzamil-sensitive absorptive flux of Reissner's membrane mediated by a highly Na+-selective channel that has several key characteristics in common with ???-ENaC. The amiloride-sensitive pathway therefore absorbs only Na+ in this epithelium and does not provide a parasensory K+ efflux route from scala media.

2011-01-01

122

Urogenital Epithelial Cells as Simple Markers of Estrogen Response in Infants: Methods and Applications  

PubMed Central

Exposure to estrogen-mimicking chemicals during critical periods of development, such as infancy, may have adverse effects. However, these effects can be difficult to characterize in most epidemiologic studies. For example, growth of reproductive organs may be susceptible to estrogenic chemicals, but measuring it requires skilled ultrasound examination; timing of pubertal onset may be altered, but observing it requires long-term follow up. To address the need for a simple marker of response to estrogenic exposures in infants, we propose a novel application of a classic marker of estrogen response in adult women: cytological evaluation of urogenital epithelial cells. In this cross-sectional study of 34 female and 41 male infants, we demonstrate that epithelial cells can be obtained from swabs of the vaginal introitus (females) and urethral meatus (males), as well as from spun urine, and that these cells respond to differential estrogenic conditions, as indicated by the relative abundance of the superficial epithelial cell type. To model varying estrogen exposure, we sampled from infants who were either newborn (highly exposed to maternal estrogens), or 12 weeks old (12W) (negligibly exposed to estrogen). Newborns had a higher percentage of superficial cells (%S), as compared to 12W (mean ± standard error: 8.3 ± 1.8 vs. 0.9 ± 0.2) (p < 0.01), consistent with an estrogen response. This difference in %S from newborn to 12W was observed similarly for swab (-7.6 ± 1.7) and urine (-7.3 ± 2.6) specimens and for males (-9.6 ± 2.9) and females (-5.2 ± 2.1). Examination of urogenital epithelial cells can successfully demonstrate estrogen response in both sexes, using cell specimens collected from either swab or urine sampling. In future studies, this simple, non-invasive method may be applied to assess whether estrogen-mimicking chemicals produce an estrogenic response in infants.

Adgent, Margaret A.; Flake, Gordon P.; Umbach, David M.; Stallings, Virginia A.; Bernbaum, Judy C.; Rogan, Walter J.

2013-01-01

123

Vaginal discharge  

MedlinePLUS

... infections and sexually transmitted infections (STIs) Trichomoniasis Vaginal yeast infection ... is never recommended. Use an over-the-counter yeast infection treatment cream or vaginal suppository, if you ...

124

Desmosome dynamics in migrating epithelial cells requires the actin cytoskeleton  

PubMed Central

Re-modeling of epithelial tissues requires that the cells in the tissue rearrange their adhesive contacts in order to allow cells to migrate relative to neighboring cells. Desmosomes are prominent adhesive structures found in a variety of epithelial tissues that are believed to inhibit cell migration and invasion. Mechanisms regulating desmosome assembly and stability in migrating cells are largely unknown. In this study we established a cell culture model to examine the fate of desmosomal components during scratch wound migration. Desmosomes are rapidly assembled between epithelial cells at the lateral edges of migrating cells and structures are transported in a retrograde fashion while the structures become larger and mature. Desmosome assembly and dynamics in this system are dependent on the actin cytoskeleton prior to being associated with the keratin intermediate filament cytoskeleton. These studies extend our understanding of desmosome assembly and provide a system to examine desmosome assembly and dynamics during epithelial cell migration.

Roberts, Brett J.; Pashaj, Anjeza; Johnson, Keith R.; Wahl, James K.

2011-01-01

125

Filifactor alocis interactions with gingival epithelial cells  

PubMed Central

An association between the gram-positive anaerobe Filifactor alocis and periodontal disease has recently emerged; however, possible pathogenic mechanisms have not been investigated. In this study we examined the responses of primary cultures of gingival epithelial cells (GECs) to infection with F. alocis. Secretion of the proinflammatory cytokines IL-1?, IL-6 and TNF-? from GECs was stimulated by F. alocis infection. F. alocis also induced apoptosis in GECs through pathways that involved caspase-3 but not caspase-9. Apoptosis was coincident with inhibition of MEK (MAPK kinase) activation. These results show that F. alocis has characteristics in common with established periodontal pathogens and has the potential to contribute to periodontal tissue destruction.

Moffatt, Catherine E.; Whitmore, Sarah E.; Griffen, Ann L.; Leys, Eugene J.; Lamont, Richard J.

2011-01-01

126

Collective Epithelial Migration and Cell Rearrangements Drive Mammary Branching Morphogenesis  

PubMed Central

Summary Epithelial organs are built through the movement of groups of interconnected cells. We observed cells in elongating mammary ducts reorganize into a multilayered epithelium, migrate collectively, and rearrange dynamically, all without forming leading cellular extensions. Duct initiation required proliferation, Rac, and myosin light-chain kinase, whereas repolarization to a bilayer depended on Rho kinase. We observed that branching morphogenesis results from the active motility of both luminal and myoepithelial cells. Luminal epithelial cells advanced collectively, whereas myoepithelial cells appeared to restrain elongating ducts. Significantly, we observed that normal epithelium and neoplastic hyperplasias are organized similarly during morphogenesis, suggesting common mechanisms of epithelial growth.

Ewald, Andrew J.; Brenot, Audrey; Duong, Myhanh; Chan, Bianca S.; Werb, Zena

2009-01-01

127

Can bone marrow cells give rise to cornea epithelial cells?  

PubMed

The corneal epithelium plays a critical role in maintaining the cornea's transparency and its avascularity. Severe damage to the limbal region results in serious problems with the corneal surface such as persistent epithelial defects, conjunctivalisation with vascularisation, keratinisation, scarring, etc. with associated profound visual loss. In order to rescue such damaged ocular surfaces, corneal epithelial stem cells were used to reconstruct artificial corneas by employing tissue engineering method. This procedure, however, requires a large limbal graft from the healthy eye and it is not possible in patients who have bilateral lesions. Therefore we should find other autologous cells as a source of cells for the reconstruction of the corneal surface. c-kit+ enriched bone marrow stem cells can give rise to different epithelial cells. So we hypothesize that this might apply to the cornea as well. Cultured cell sheets composed of autologous c-kit+ enriched bone marrow stem cells may be used to reconstruct corneal surfaces and can restore vision in patients with bilateral severe disorders of the ocular surface. PMID:18495366

Liu, Yuan; Wang, Xinwen; Jin, Yan

2008-05-21

128

Epithelial Cell Rests of Malassez Contain Unique Stem Cell Populations Capable of Undergoing Epithelial-Mesenchymal Transition  

PubMed Central

The epithelial cell rests of Malassez (ERM) are odontogenic epithelial cells located within the periodontal ligament matrix. While their function is unknown, they may support tissue homeostasis and maintain periodontal ligament space or even contribute to periodontal regeneration. We investigated the notion that ERM contain a subpopulation of stem cells that could undergo epithelial–mesenchymal transition and differentiate into mesenchymal stem-like cells with multilineage potential. For this purpose, ERM collected from ovine incisors were subjected to different inductive conditions in vitro, previously developed for the characterization of bone marrow mesenchymal stromal/stem cells (BMSC). We found that ex vivo-expanded ERM expressed both epithelial (cytokeratin-8, E-cadherin, and epithelial membrane protein-1) and BMSC markers (CD44, CD29, and heat shock protein-90?). Integrin ?6/CD49f could be used for the enrichment of clonogenic cell clusters [colony-forming units-epithelial cells (CFU-Epi)]. Integrin ?6/CD49f-positive-selected epithelial cells demonstrated over 50- and 7-fold greater CFU-Epi than integrin ?6/CD49f-negative cells and unfractionated cells, respectively. Importantly, ERM demonstrated stem cell-like properties in their differentiation capacity to form bone, fat, cartilage, and neural cells in vitro. When transplanted into immunocompromised mice, ERM generated bone, cementum-like and Sharpey's fiber-like structures. Additionally, gene expression studies showed that osteogenic induction of ERM triggered an epithelial–mesenchymal transition. In conclusion, ERM are unusual cells that display the morphological and phenotypic characteristics of ectoderm-derived epithelial cells; however, they also have the capacity to differentiate into a mesenchymal phenotype and thus represent a unique stem cell population within the periodontal ligament.

Xiong, Jimin; Mrozik, Krzysztof; Gronthos, Stan

2012-01-01

129

A rare germ-cell tumor site: vaginal endodermal sinus tumor  

Microsoft Academic Search

Malignant germ-cell tumors (MGCT) are rare tumors of childhood accounting for less than 3% of pediatric malignancies. Endodermal sinus tumor (EST) forms the most common histologic subtype of MGCT. The vagina is an extremely rare site for GCTs. A 9-month-old female was admitted with a short history of vaginal bleeding, a mass protruding from the vagina, and difficulty in passing

M. Arora; R. Shrivastav; M. Jaiprakash

2002-01-01

130

Serial culturing of human bronchial epithelial cells derived from biopsies  

Microsoft Academic Search

Summary  In the present study we describe the establishment of serial cultures of human bronchial epithelial cells derived from biopsies\\u000a obtained by fiberoptic bronchoscopy. The cell cultures were initiated from small amounts of material (2 mm forceps biopsies)\\u000a using either explants or epithelial cell suspensions in combination with a feeder-layer technique. The rate of cell proliferation\\u000a and the number of passages

Petra M. de Jong; Marianne A. J. A. van Sterkenburg; Johanna A. Kempenaar; Joop H. Dijkman; Maria Ponec

1993-01-01

131

Induction of Epithelial-Mesenchymal Transition in Alveolar Epithelial Cells by Transforming Growth Factor-?1  

PubMed Central

The hallmark of idiopathic pulmonary fibrosis (IPF) is the myofibroblast, the cellular origin of which in the lung is unknown. We hypothesized that alveolar epithelial cells (AECs) may serve as a source of myofibroblasts through epithelial-mesenchymal transition (EMT). Effects of chronic exposure to transforming growth factor (TGF)-?1 on the phenotype of isolated rat AECs in primary culture and a rat type II cell line (RLE-6TN) were evaluated. Additionally, tissue samples from patients with IPF were evaluated for cells co-expressing epithelial (thyroid transcription factor (TTF)-1 and pro-surfactant protein-B (pro-SP-B), and mesenchymal (?-smooth muscle actin (?-SMA)) markers. RLE-6TN cells exposed to TGF-?1 for 6 days demonstrated increased expression of mesenchymal cell markers and a fibroblast-like morphology, an effect augmented by tumor necrosis factor-? (TNF-?). Exposure of rat AECs to TGF-?1 (100 pmol/L) resulted in increased expression of ?-SMA, type I collagen, vimentin, and desmin, with concurrent transition to a fibroblast-like morphology and decreased expression of TTF-1, aquaporin-5 (AQP5), zonula occludens-1 (ZO-1), and cytokeratins. Cells co-expressing epithelial markers and ?-SMA were abundant in lung tissue from IPF patients. These results suggest that AECs undergo EMT when chronically exposed to TGF-?1, raising the possibility that epithelial cells may serve as a novel source of myofibroblasts in IPF.

Willis, Brigham C.; Liebler, Janice M.; Luby-Phelps, Katherine; Nicholson, Andrew G.; Crandall, Edward D.; du Bois, Roland M.; Borok, Zea

2005-01-01

132

Myeloma light chains induce epithelial-mesenchymal transition in human renal proximal tubule epithelial cells  

Microsoft Academic Search

Background. To determine the role of epithelial- mesenchymal transition (EMT) as a potential mechanism contributing to the characteristic tubulointerstitial renal fi- brosisinmultiplemyeloma,weexaminedwhethermyeloma light chains (LCs) directly induce EMT in human renal proximal tubule epithelial cells (PTECs). Methods. As positive controls we used TGF-?1 and cy- closporine A (CsA), two agents known to induce EMT in PTECs. Human LCs were isolated

Min Li; Kathleen S. Hering-Smith; Eric E. Simon; Vecihi Batuman

133

Separation and Characterization of Epithelial and Mesenchymal-like Murine Mammary Tumor Cells Reveals Epithelial Cell Differentiation Plasticity and Enhanced Tumorigenicity of Epithelial-enriched Tumor Cells.  

PubMed

Tumors are composed of heterogeneous populations of cells including tumor-initiating cells (TICs) and metastatic precursors. While the origin of these cells is unknown, there is evidence that tumor cells can transdifferentiate from an epithelial to a mesenchymal phenotype, a property referred to as epithelial-to-mesenchymal transition (EMT). This cellular plasticity may explain the heterogeneous nature of tumors and differences in the tumorigenic and invasive properties of cells. Understanding the origin of these cells and the contribution of external factors that influence the acquisition of cellular properties is critical for the development of therapeutics to eradicate cancer. In this study, we show that primary murine tumor cells harvested from FVB/N Tg (MMTV/Neu) spontaneous mammary tumors possess differentiation plasticity and can be enriched to be epithelial or mesenchymal-like using selected culture media conditions, and we show evidence of EMT in a clonal population of primary epithelial tumor cells when cultured in fibroblast growth factor-1 (FGF-1) or transforming growth factor-? (TGF-?). We also determined that in contrast to the identification of mesenchymal-like tumor cells as TICs in orthotopic xenograph models of tumorigenicity, epithelial-enriched murine mammary tumor cells were more tumorigenic as compared to mesenchymal-enriched cells when transplanted back subcutaneously into syngeneic immune competent mice. Together, these data suggest that EMT plasticity can be induced in primary murine mammary tumor cells, and that tumorigenicity of epithelial or mesenchymal-like cells may be influenced by factors such as the site of tumor inoculation or the immune state of the host (xenogenic immune compromised versus syngeneic immune competent). PMID:22237886

Palen, Katie A; Jing, Weiqing; Weber, James J; Tilkens, Sara B; Chan, Andrew M; Johnson, Bryon D; Gershan, Jill A

2012-01-12

134

Stimulating Vaginal Repair in Rats Through Skeletal Muscle-Derived Stem Cells Seeded on Small Intestinal Submucosal Scaffolds  

PubMed Central

OBJECTIVES Grafts are used for vaginal repair after prolapse, but their use to carry stem cells to regenerate vaginal tissue has not been reported. In this study, we investigated whether 1) muscle-derived stem cells (MDSC) grown on small intestinal submucosa (SIS) generate smooth-muscle cells (SMC) in vitro and upon implantation in a rat model of vaginal defects; 2) express markers applicable to the in-vivo detection of vaginal endogenous stem cells; and 3) stimulate the repair of the vagina. METHODS Mouse MDSC grown on monolayer, SIS, or polymeric mesh, were tested for cell differentiation by immunocytochemistry, Western blot and real-time polymerase chain reaction (PCR). Stem cell markers were screened by DNA microarrays followed by real-time PCR, immunocytochemistry, and Western blot. Rats that underwent hysterectomy and partial vaginectomy were left as such or implanted in the vagina with 4’,6-Diamidino-2-Phenylindole (DAPI)–labeled MDSC on SIS, or SIS without MDSC, immunosuppressed, and killed at 2–8 weeks. Immunofluorescence, hematoxylin-eosin, and Masson trichrome were applied to tissue sections. RESULTS Muscle-derived stem cell cultures on monolayer and on scaffolds differentiate into SMC, as shown by ?-smooth muscle actin (ASMA), calponin, and smoothelin markers. Muscle-derived stem cells express embryonic stem cell markers Oct-4 and nanog. Dual DAPI/ASMA fluorescence indicated MDSC conversion to SMC. Muscle-derived stem cells/SIS stimulated vaginal tissue repair, including keratin-5 positive epithelium formation and prevented fibrosis at 4 and 8 weeks. Oct-4+ putative endogenous stem cells were identified. CONCLUSION Muscle-derived stem cells/SIS implants stimulate vaginal tissue repair in the rat, thus autologous MDSC on scaffolds may be a promising approach for the treatment of vaginal repair.

Ho, Matthew H.; Heydarkhan, Sanaz; Vernet, Dolores; Kovanecz, Istvan; Ferrini, Monica G.; Bhatia, Narender N.; Gonzalez-Cadavid, Nestor F.

2011-01-01

135

Tracking the intermediate stages of epithelial-mesenchymal transition in epithelial stem cells and cancer.  

PubMed

Epithelial-mesenchymal transition (EMT) is an essential developmental program that becomes reactivated in adult tissues to promote the progression of cancer. EMT has been largely studied by examining the beginning epithelial state or the ending mesenchymal state without studying the intermediate stages. Recent studies using trophoblast stem (TS) cells paused in EMT have defined the molecular and epigenetic mechanisms responsible for modulating the intermediate "metastable" stages of EMT. Targeted inactivation of MAP3K4, knockdown of CBP, or overexpression of SNAI1 in TS cells induced similar metastable phenotypes. These TS cells exhibited epigenetic changes in the histone acetylation landscape that cause loss of epithelial maintenance while preserving self-renewal and multipotency. A similar phenotype was found in claudin-low breast cancer cells with properties of EMT and stemness. This intersection between EMT and stemness in TS cells and claudin-low metastatic breast cancer demonstrates the usefulness of developmental EMT systems to understand EMT in cancer. PMID:21862874

Jordan, Nicole Vincent; Johnson, Gary L; Abell, Amy N

2011-09-01

136

Fusion between Hematopoietic and Epithelial Cells in Adult Human Intestine  

PubMed Central

Following transplantation of hematopoietic lineage cells, genetic markers unique to the transplanted cells have been detected in non-hematopoietic recipient cells of human liver, vascular endothelium, intestinal epithelium and brain. The underlying mechanisms by which this occurs are unclear. Evidence from mice suggests it is due in part to fusion between cells of hematopoietic and non-hematopoietic origins; however, direct evidence for this in humans is scant. Here, by quantitative and statistical analysis of X- and Y-chromosome numbers in epithelial and non-epithelial intestinal cells from gender-mismatched hematopoietic cell transplant patients, we provide evidence that transplanted cells of the hematopoietic lineage incorporate into human intestinal epithelium through cell fusion. This is the first definitive identification of cell fusion between hematopoietic cells and any epithelial cell type in humans, and provides the basis for further understanding the physiological and potential pathological consequences of cell fusion in humans.

Silk, Alain D.; Gast, Charles E.; Davies, Paige S.; Fakhari, Farnaz D.; Vanderbeek, Gretchen E.; Mori, Motomi; Wong, Melissa H.

2013-01-01

137

Fibronectin Expression Modulates Mammary Epithelial Cell Proliferation during Acinar Differentiation  

Microsoft Academic Search

The mammary gland consists of a polarized epithelium surrounded by a basement membrane matrix that forms a series of branching ducts ending in hollow, sphere-like acini. Essential roles for the epithelial basement membrane during acinar differentiation, in particular laminin and its integrin receptors, have been identified using mammary epithelial cells cultured on a reconstituted basement membrane. Contributions from fibronectin, which

Courtney M. Williams; Adam J. Engler; R. Daniel Slone; Leontine L. Galante; Jean E. Schwarzbauer

2008-01-01

138

Primary airway epithelial cell culture from lung transplant recipients  

Microsoft Academic Search

Long-term survival in lung transplantation is limited by the development of obliterative bronchiolitis, a condition characterised by inflammation, epithelial injury, fibroproliferation and obliteration of bronchioles leading to airflow obstruction. To investigate the role of the bronchial epithelium in the pathogenesis of obliterative bronchiolitis the current study aimed to establish primary bronchial epithelial cell cultures (PBEC) from lung allografts. Four to

I. A. Forrest; D. M. Murphy; C. Ward; D. Jones; G. E. Johnson; L. Archer; F. K. Gould; T. E. Cawston; J. L. Lordan; P. A. Corris

2005-01-01

139

Differentiation capacity of epithelial cells in the sponge Suberites domuncula  

Microsoft Academic Search

Sponges (phylum Porifera) represent the oldest metazoans. Their characteristic metazoan adhesion molecules and transcription factors enable them to establish a complex “Bauplan”; three major differentiated cell types (epithelial cells, skeletal cells\\/sclerocytes, and contractile cells) can be distinguished. Since no molecular markers are as yet available to distinguish these somatic cells or the corresponding embryonic cells from which they originate, we

Heinz C. Schröder; Sanja Perovi?-Ottstadt; Matthias Wiens; Renato Batel; Isabel M. Müller; Werner E. G. Müller

2004-01-01

140

Growth of corneal epithelial cells over in situ therapeutic contact lens after simple limbal epithelial transplantation (SLET).  

PubMed

An 11-year-old boy underwent simple limbal epithelial transplantation (SLET) from the healthy right eye to his left eye for total limbal stem cell deficiency. One month later, corneal surface epithelialised and whitish plaques overlying the transplants were seen inferiorly. Those plaques were adherent to the surface of the contact lens and underlying corneal surface had smooth elevations. Similar findings were noted in a 23-year man following cyanoacrylate glue application for corneal perforation. On histological and immunohistochemical analysis, cells lining the contact lenses were identified as corneal epithelial cells. These cases illustrate epithelial cell growth on the contact lens and epithelial hyperplasia on corresponding surface of the cornea. Exorbitant proliferation of the epithelial cells may be owing to young age; therefore, early contact lens removal after SLET in young age, can possibly avoid epithelial hyperplasia. This also reiterates the possibility of using contact lens as a scaffold to grow epithelial cells. PMID:23814196

Bhalekar, Swapnil; Sangwan, Virender S; Basu, Sayan

2013-06-27

141

Regulation of lung fibroblast tropoelastin expression by alveolar epithelial cells.  

PubMed

Epithelial-mesenchymal interactions are of critical importance during tissue morphogenesis and repair. Although the cellular and molecular aspects of many of these interactions are beginning to be understood, the ability of epithelial cells to regulate fibroblast interstitial matrix production has not been extensively studied. We report here that cultured alveolar epithelial cells are capable of modulating the expression of tropoelastin, the soluble precursor of the interstitial lung matrix component elastin, by lung fibroblasts. Phorbol ester-stimulated alveolar epithelial cells secrete a soluble factor that causes a time- and dose-dependent repression of lung fibroblast tropoelastin mRNA expression. This alveolar epithelial cell-mediated repressive activity is specific for tropoelastin, is effective on lung fibroblasts from multiple stages of development, and acts at the level of transcription. Partial characterization of the repressive activity indicates it is an acid-stable, pepsin-labile protein. Gel fractionation of alveolar epithelial cell conditioned medium revealed two peaks of activity with relative molecular masses of approximately 25 and 50 kDa. These data support a role for epithelial cells in the regulation of fibroblast interstitial matrix production. PMID:9458800

Mariani, T J; Dunsmore, S E; Li, Q; Ye, X; Pierce, R A

1998-01-01

142

Airway Epithelial IL15 Transforms Monocytes into Dendritic Cells  

Microsoft Academic Search

IL-15 has recently been shown to induce the differentiation of func- tional dendritic cells (DCs) from human peripheral blood mono- cytes.SinceDCslay inclose proximity toepithelialcells inthe airway mucosa, we investigated whether airway epithelial cells release IL-15 in response to inflammatory stimuli and thereby induce differ- entiation and maturation of DCs. Alveolar (A549) and bronchial (BEAS-2B) epithelial cells produced IL-15 spontaneously and

Nicolas Regamey; Carolina Obregon; Sylvie Ferrari-Lacraz; Coretta van Leer; Marc Chanson; Laurent P. Nicod; Thomas Geiser

143

GROWTH OF POSTEMBRYONIC SKIN EPITHELIAL CELLS ON COLLAGEN GELS  

Microsoft Academic Search

A procedure to extract and to purify acid-soluble collagen from rabbit skin and a technique to prepare a semi-transparent collagen gel to support growth of postembryonic rabbit, mouse, and human skin epithelial cells are presented. When a suspension of trypsin-released epithelial cells is plated on a collagen gel, the reorganization and growth of cells takes place in 3 stages. In

Marvin A. Karasek; Mary Ellen Charlton

1971-01-01

144

Genetics and epithelial cell dysfunction in cystic fibrosis  

SciTech Connect

This book examines the advances being made in the study of the physiology, cell biology, and molecular genetics of cystic fibrosis. Emphasis is placed on various areas of research that involve epithelial cells (e.g., the CF-specific phenotypes exhibited by epithelial cells, abnormalities in epithelium ion transport, chloride channel regulation in CF epithelial.) Coverage is presented on the current status of CF, including data on the incidence of the disease, its mode of inheritance, chromosomal localization, genetic heterogeneity, and screening and management.

Riordan, J.R.; Buchwald, M.

1987-01-01

145

Cell Cycle Arrest by Kynurenine in Lens Epithelial Cells  

PubMed Central

Purpose Indolemine 2,3-dioxygenase (IDO)-mediated oxidation of tryptophan produces kynurenines (KYNs), which may play a role in cataract formation. The molecular mechanisms by which KYNs cause cellular changes are poorly understood. The effects of KYNs on mouse lens epithelial cells by overexpression of human IDO were investigated. Methods Lens epithelial cells (mLECs) derived from human IDO-overexpressing hemizygous transgenic (hemTg) and wild-type (Wt) mice were used. IDO activity was measured by quantifying kynurenine (KYN) by HPLC. KYN-mediated protein modifications were detected by immunocytochemistry and measured by ELISA. Cell proliferation and apoptosis were measured with commercially available kits. Cell distribution between cell cycle phases was examined with flow cytometric analysis. Immunoprecipitation followed by LC/MS was used to identify kynurenine-modified proteins. Results mLECs derived from hemTg animals exhibited considerable IDO immunoreactivity and enzyme activity, which were barely detectable in Wt mLECs. KYN and KYN-mediated protein modification were detected in hemTg but not in Wt mLECs; the modified proteins were myosin II and ?/?-actin. HemTg mLECs displayed reduced viability and proliferation. Cell cycle analysis of hemTg mLEC cultures showed approximately a twofold increase in cells at G2/M or in both phases, relative to Wt mLECs. Blocking IDO activity with 1-methyl-d,l-tryptophan in hemTg mLECs prevented KYN formation, KYN-mediated protein modification, and G2/M arrest. Conclusions Excess IDO activity in mLECs results in KYN production, KYN-mediated modification of myosin II and ?/?-actin, and cell cycle perturbation. Modification of myosin II and ?-actin by KYN may interfere with cytokinesis, leading to defective epithelial cell division and thus a decreased number of fiber cells.

Mailankot, Maneesh; Smith, Dawn; Howell, Scott; Wang, Benlian; Jacobberger, James W.; Stefan, Tammy; Nagaraj, Ram H.

2008-01-01

146

Transcriptional Responses of Candida albicans to Epithelial and Endothelial Cells? †  

PubMed Central

Candida albicans interacts with oral epithelial cells during oropharyngeal candidiasis and with vascular endothelial cells when it disseminates hematogenously. We set out to identify C. albicans genes that govern interactions with these host cells in vitro. The transcriptional response of C. albicans to the FaDu oral epithelial cell line and primary endothelial cells was determined by microarray analysis. Contact with epithelial cells caused a decrease in transcript levels of genes related to protein synthesis and adhesion, whereas contact with endothelial cells did not significantly influence any specific functional category of genes. Many genes whose transcripts were increased in response to either host cell had not been previously characterized. We constructed mutants with homozygous insertions in 22 of these uncharacterized genes to investigate their function during host-pathogen interaction. By this approach, we found that YCK2, VPS51, and UEC1 are required for C. albicans to cause normal damage to epithelial cells and resist antimicrobial peptides. YCK2 is also necessary for maintenance of cell polarity. VPS51 is necessary for normal vacuole formation, resistance to multiple stressors, and induction of maximal endothelial cell damage. UEC1 encodes a unique protein that is required for resistance to cell membrane stress. Therefore, some C. albicans genes whose transcripts are increased upon contact with epithelial or endothelial cells are required for the organism to damage these cells and withstand the stresses that it likely encounters during growth in the oropharynx and bloodstream.

Park, Hyunsook; Liu, Yaoping; Solis, Norma; Spotkov, Joshua; Hamaker, Jessica; Blankenship, Jill R.; Yeaman, Michael R.; Mitchell, Aaron P.; Liu, Haoping; Filler, Scott G.

2009-01-01

147

Puromycin aminonucleoside suppresses integrin expression in cultured glomerular epithelial cells.  

PubMed

Puromycin aminonucleoside (PAN)-induced nephrosis is a well-described model of human idiopathic nephrotic syndrome, but the mechanism of PAN's effect is not completely understood. Because PAN injection into rats results in retraction of glomerular epithelial cell foot processes and glomerular epithelial cell detachment, it was hypothesized that PAN might alter the contacts between these cells and the glomerular basement membrane. The major integrin expressed by glomerular epithelial cells is alpha3beta1, which mediates attachment of these cells to extracellular matrix proteins including type IV collagen. T-SV 40 immortalized human glomerular epithelial cells were used to study PAN's effects on alpha3beta1 expression, as well as that of podocalyxin and the slit diaphragm component ZO-1. Glomerular epithelial cells were seeded into plastic flasks and allowed to attach and proliferate for 48 h. The cells were then incubated for another 48 h in media containing 0, 0.5, or 5.0 microg/ml PAN. PAN exposure resulted in dose-dependent decreases in alpha3 and beta1 expression, both at the protein level and at the mRNA level. This was accompanied by a significant decrease in the adhesion of glomerular epithelial cells to type IV collagen. PAN did not affect ZO-1 protein expression. Treatment with PAN increased the expression of podocalyxin at the protein and mRNA levels. Reduced glomerular epithelial cell expression of alpha3beta1 integrins and impaired adhesion to type IV collagen may contribute to the glomerular epithelial cell detachment from glomerular basement membrane seen in the PAN nephrosis model. PMID:11274237

Krishnamurti, U; Zhou, B; Fan, W W; Tsilibary, E; Wayner, E; Kim, Y; Kashtan, C E; Michael, A

2001-04-01

148

The Properties of Retinal Pigment Epithelial Cells in Proliferative Vitreoretinopathy Compared with Cultured Retinal Pigment Epithelial Cells  

Microsoft Academic Search

Retinal pigment epithelial (RPE) cells, which proliferate and dedifferentiate under several pathological conditions, and cultured RPE cells have been considered a good model for comparison. In this investigation, we compared the properties of RPE cells in proliferative vitreoretinopathy with that of cultured human RPE cells. mRNAs of RPE cells from patients with proliferative vitreoretinopathy and from cultured human RPE cells

TOSHIAKI ABE; YUSUF K. DURLU; MAKOTO TAMAI

1996-01-01

149

Role for Dendritic Cells in Immunoregulation during Experimental Vaginal Candidiasis  

Microsoft Academic Search

Vulvovaginal candidiasis (VVC) caused by the commensal organism Candida albicans remains a significant problem among women of childbearing age, with protection against and susceptibility to infection still poorly understood. While cell-mediated immunity by CD4 Th1-type cells is protective against most forms of mucosal candidiasis, no protective role for adaptive immunity has been identified against VVC. This is postulated to be

Dana M. LeBlanc; Melissa M. Barousse; Paul L. Fidel

2006-01-01

150

Multipotent Capacity of Immortalized Human Bronchial Epithelial Cells  

Microsoft Academic Search

While the adult murine lung utilizes multiple compartmentally restricted progenitor cells during homeostasis and repair, much less is known about the progenitor cells from the human lung. Translating the murine stem cell model to humans is hindered by anatomical differences between species. Here we show that human bronchial epithelial cells (HBECs) display characteristics of multipotent stem cells of the lung.

Oliver Delgado; Aadil A. Kaisani; Monica Spinola; Xian-Jin Xie; Kimberly G. Batten; John D. Minna; Woodring E. Wright; Jerry W. Shay

2011-01-01

151

Characteristics of Helicobacter pylori attachmentto human primary antral epithelial cells  

Microsoft Academic Search

Conventional cell lines are commonly used to study infection characteristics of the human gastric pathogen Helicobacter pylori. We sought to investigate bacterial attachment to human antral primary epithelial cells, a cell model that more closely resembles the human stomach than transformed cell lines. Primary cells were infected for 24 and 48 h with H. pylori. Morphological appearance of both the pathogen and

Ursula Heczko; Valerie C Smith; R Mark Meloche; Alison M. J Buchan; B. Brett Finlay

2000-01-01

152

Crystal violet staining to quantify Candida adhesion to epithelial cells.  

PubMed

In vitro studies of adhesion capability are essential to characterise the virulence of Candida species. However, the assessment of adhesion by traditional methods is time-consuming. The aim of the present study is the development of a simple methodology using crystal violet staining to quantify in vitro adhesion of different Candida species to epithelial cells. The experiments are performed using Candida albicans (ATCC 90028), C. glabrata (ATCC 2001), C. parapsilosis (ATCC 22019) and C. tropicalis (ATCC 750). A human urinary bladder epithelial cell line (TCC-SUP) is used. Yeast and epithelial cells were stained with crystal violet, epithelial cells were then destained using intermediate washing, and the dye in the yeast cells was extracted with acetic acid. The method was validated for the different Candida reference species by comparison with traditional microscope observation and enumeration. The method was then used to assess Candida adhesion to epithelial cells and also to silicone. For all Candida spp. high correlation values (r2= 0.9724-0.9997) between the number of adherent yeasts (microscope enumeration) and absorbance values were obtained for an inoculum concentration >10(6) cells/mL. The proposed technique was easy to perform and reproducible, enabling the determination of adhesion ability of Candida species to an epithelial cell line. PMID:20973406

Negri, M; Gonçalves, V; Silva, S; Henriques, M; Azeredo, J; Oliveira, R

2010-01-01

153

Epithelial stem cell mutations that promote squamous cell carcinoma metastasis  

PubMed Central

Squamous cell carcinomas (SCCs) originate in stratified epithelia, with a small subset becoming metastatic. Epithelial stem cells are targets for driver mutations that give rise to SCCs, but it is unknown whether they contribute to oncogenic multipotency and metastasis. We developed a mouse model of SCC by targeting two frequent genetic mutations in human SCCs, oncogene KrasG12D activation and Smad4 deletion, to mouse keratin 15–expressing (K15+) stem cells. We show that transgenic mice developed multilineage tumors, including metastatic SCCs. Among cancer stem cell–enriched (CSC-enriched) populations, those with increased side population (SP) cells correlated with epithelial-mesenchymal transition (EMT) and lung metastasis. We show that microRNA-9 (miR-9) contributed to SP expansion and metastasis, and miR-9 inhibition reduced the number of SP cells and metastasis. Increased miR-9 was detected in metastatic human primary SCCs and SCC metastases, and miR-9–transduced human SCC cells exhibited increased invasion. We identified ?-catenin as a predominant miR-9 target. Increased miR-9 in human SCC metastases correlated with ?-catenin loss but not E-cadherin loss. Our results demonstrate that stem cells with KrasG12D activation and Smad4 depletion can produce tumors that are multipotent and susceptible to EMT and metastasis. Additionally, tumor initiation and metastatic properties of CSCs can be uncoupled, with miR-9 regulating the expansion of metastatic CSCs.

White, Ruth A.; Neiman, Jill M.; Reddi, Anand; Han, Gangwen; Birlea, Stanca; Mitra, Doyel; Dionne, Laikuan; Fernandez, Pam; Murao, Kazutoshi; Bian, Li; Keysar, Stephen B.; Goldstein, Nathaniel B.; Song, Ningjing; Bornstein, Sophia; Han, Zheyi; Lu, Xian; Wisell, Joshua; Li, Fulun; Song, John; Lu, Shi-Long; Jimeno, Antonio; Roop, Dennis R.; Wang, Xiao-Jing

2013-01-01

154

HIV is inactivated after transepithelial migration via adult oral epithelial cells but not fetal epithelial cells  

PubMed Central

Oral transmission of human immunodeficiency virus (HIV) in adult populations is rare. However, HIV spread across fetal/neonatal oropharyngeal epithelia could be important in mother-to-child transmission. Analysis of HIV transmission across polarized adult and fetal oral epithelial cells revealed that HIV transmigrates through both adult and fetal cells. However, only virions that passed through the fetal cells – and not those that passed through the adult cells – remained infectious. Analysis of expression of anti-HIV innate proteins beta-defensins 2 and 3, and secretory leukocyte protease inhibitor in adult, fetal, and infant oral epithelia showed that their expression is predominantly in the adult oral epithelium. Retention of HIV infectivity after transmigration correlated inversely with the expression of these innate proteins. Inactivation of innate proteins in adult oral keratinocytes restored HIV infectivity. These data suggest that high-level innate protein expression may contribute to the resistance of the adult oral epithelium to HIV transmission.

Tugizov, Sharof M.; Herrera, Rossana; Veluppillai, Piri; Greenspan, Deborah; Soros, Vanessa; Greene, Warner C.; Levy, Jay A.; Palefsky, Joel M.

2010-01-01

155

Epithelial cell characteristics of cultured human limbal explants  

PubMed Central

Aim: To determine the immunohistochemical characteristics of putative corneal epithelial stem cells remaining on limbal explants maintained in culture. Methods: Human limbal explant cultures were generated from 25 residual corneoscleral donor rims following penetrating keratoplasty. Serial sections of these explants were studied using immunohistochemical techniques with a panel of antibodies, on day 0 and 1, 2, and 3 weeks. Results: The number of epithelial cells expressing cytokeratin 19 and vimentin increased with duration in culture, while the number of cells expressing cytokeratin 3 decreased. Connexin 43 expression was lost by 1 week in culture. p63 was expressed by cells that had migrated around the explant and the number of p63 positive cells decreased with longer duration in culture. The explants were initially negative for Ki67, but the epithelial cells were positive at 1 week, and expression of Ki67 was progressively lost with increasing duration in culture. The initial uniform staining of the epithelium for epidermal growth factor receptor and ? enolase remained unchanged at 3 weeks. Conclusions: There is an expansion of less differentiated (cytokeratin 3 negative and CK19/vimentin positive) epithelial cells on corneoscleral explants maintained in culture for 3 weeks. The pattern of expression of p63 noted in this study does not support the suggestion that it is a marker of limbal stem cells. The decline in p63 and Ki67 expression among the epithelial cells of the cultured explant button implies that as the epithelial sheet outgrowing from the explant button reaches confluence, the proliferative status of the cells remaining on the explant button declines. These findings are of clinical relevance as explants of limbal tissue are used in limbal stem cell transplantation. There is no information available to date on the fate of epithelial cells on such explants. This study provides some insight into this and suggests that an expansion of the stem cell pool or its progeny may occur in limbal explants.

Joseph, A; Powell-Richards, A O R; Shanmuganathan, V A; Dua, H S

2004-01-01

156

Neutrophils rescue gingival epithelial cells from bacterial-induced apoptosis  

PubMed Central

In the pathogenesis of chronic inflammatory periodontal disease, neutrophils are recognized as a major cellular component from the histopathology of the periodontal lesion around teeth and from clinical cases where absence or dysfunction of neutrophils results in major periodontal destruction. Neutrophils are recruited in vast numbers into the gingival crevice during periodontal inflammation, attracted by microbial plaque chemoattractants and chemokines released following microbial perturbation of gingival epithelial cells. Porphyromonas gingivalis, a major periodontopathogen, triggers a vast array of cellular responses in gingival epithelial cells but also induces apoptosis. We demonstrate here that neutrophils, when combined in a P. gingivalis challenge assay of epithelial cells, prevent epithelial cell apoptosis by phagocytosing P. gingivalis and later undergoing apoptosis themselves. By removing P. gingivalis by phagocytosis, neutrophils also protect the host from the harmful effects of its microbial proteases, which degrade inflammatory cytokines and other host molecules.

Galicia, Johnah C.; Benakanakere, Manjunatha R.; Stathopoulou, Panagiota G.; Kinane, Denis F.

2009-01-01

157

Cooperative Interactions During Human Mammary Epithelial Cell Immortalization.  

National Technical Information Service (NTIS)

Our laboratory has pioneered the development and use of cultured human mammary epithelial cells (HMEC) to gain information on the defects in growth control processes that allow finite lifespan HMEC to overcome all senescence barriers, reactivate telomeras...

P. Yaswen

2003-01-01

158

Role of p53 in Mammary Epithelial Cell Senescence.  

National Technical Information Service (NTIS)

Preselect ion and postselection human mammary epithelial cells (HMECs) differ greatly in replicative potential. In this report, we studied the role of p53 in senescence of preselection and postselection HMECs. Western blot analysis of early (proliferating...

G. P. Dimri

2003-01-01

159

CYTOKINE REPERTOIRE OF EPITHELIAL CELLS LINING THE HUMAN URINARY TRACT  

Microsoft Academic Search

PurposeTo examine the cytokine profile of epithelial cells lining the human urinary tract with the aim of differentiating between the constitutive and disease-related cytokine production in these tissues.

LONG HANG; BJORN WULLT; ZHENXIN SHEN; DIANA KARPMAN; CATHARINA SVANBORG

1998-01-01

160

Epithelial-mesenchymal transition in primary human bronchial epithelial cells is Smad-dependent and enhanced by fibronectin and TNF-?  

Microsoft Academic Search

BACKGROUND: Defective epithelial repair, excess fibroblasts and myofibroblasts, collagen overproduction and fibrosis occur in a number of respiratory diseases such as asthma, chronic obstructive pulmonary disease (COPD) and pulmonary fibrosis. Pathological conversion of epithelial cells into fibroblasts (epithelial-mesenchymal transition, EMT) has been proposed as a mechanism for the increased fibroblast numbers and has been demonstrated to occur in lung alveolar

Joana Câmara; Gabor Jarai

2010-01-01

161

Characteristics and EGFP expression of porcine mammary gland epithelial cells.  

PubMed

The aims of this study were to establish a porcine mammary gland epithelial (PMGE) cell line, and to determine if these PMGE cells could be maintained long-term in culture by continuous subculturing following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Primary culture of PMGE cells was achieved by outgrowth of migrating cells from the fragments of the mammary gland tissue of a lactating pig. The passage sixteen PMGE cells were transfected with EGFP gene using lipofection. The expression of Cell keratins of epithelial cells in PMGE cells was tested by immunofluorescence. ?eta-Casein gene mRNA was tested for PMGE cells by RT-PCR. The results showed that PMGE cells could form dome-like structure which looked like nipple, and the cells contained different cell types. The expression of Cell keratins demonstrated the property of epithelial cells, and the PMGE cells could express transcript encoding a ?eta-Casein protein. EGFP gene was successfully transferred into the PMGE cells, and the transfected cells could be maintained long-term in culture by continuous subculturing. In conclusion, we have established a EGFP gene transfected porcine mammary gland epithelial (ET-PMGE) cell line. PMID:20400167

Zheng, Yue-Mao; He, Xiao-Ying

2010-04-18

162

Establishment of a human endometrial cell culture system and characterization of its polarized hormone responsive epithelial cells  

Microsoft Academic Search

Uterine epithelial cells from normal human endometrium were cultured as a primary cell culture in a dual-chambered system. The epithelial cells were isolated from endometrial tissue of the proliferative phase obtained by hysterectomy. The epithelial cells were seeded on Millicell CM filters coated with the extracellular matrix Matrigel. Depending on the culture conditions, the epithelial cells formed a polarized cell

I. Classen-Linke; M. Kusche; R. Knauthe; H. M. Beier

1996-01-01

163

Isolation of two keratinocyte cell lines derived from HPV-positive dysplastic vaginal lesions.  

PubMed

Explant cultures were started from human papillomavirus (HPV)-infected genital lesions in order to isolate and propagate abnormally differentiating cells from squamous intraepithelial neoplasia. A medium with high calcium concentration was used to induce terminal differentiation of cells from surrounding normal epithelium. Two cell lines with extended life-spans were established. The UT-DEC-1 cell line was derived from an HPV-33-positive mild vaginal dysplasia (VAIN I). In cultured UT-DEC-1 cells, HPV 33 DNA was detected with Southern-blot hybridization and the polymerase chain reaction (PCR) technique. The restriction pattern of HPV 33 changed during early passages and flow cytometric analysis detected a decrease in chromosomal DNA content. HPV 33 RNA from the E6-E7 region could be amplified by PCR at late passage. UT-DEC-2 cell line was derived from an HPV-16-positive moderate vaginal dysplasia (VAIN II). HPV 16 DNA was also detected in cultured cells by the PCR technique. The senescence of normal keratinocytes and growth selection in favor of aneuploid cells was observed by flow cytometric analysis at subsequent passages. Karyotype analysis showed clonal chromosomal abnormalities in both cell lines. To date, UT-DEC-1 cells have undergone 40 and UT-DEC-2 cells 25 passages. This study shows that the isolation of HPV-infected dysplastic cells can be achieved by culturing the cells in a medium with high calcium concentration. The cell lines presented provide the opportunity of evaluating the early stages of squamous-cell carcinogenesis. PMID:1328068

Hietanen, S; Auvinen, E; Grénman, S; Lakkala, T; Sajantila, A; Klemi, P; Mäenpää, J

1992-09-30

164

Generation of a complete thymic microenvironment by MTS24+ thymic epithelial cells  

Microsoft Academic Search

The epithelial component of the thymic microenvironment is indispensable for the generation of T lymphocytes. Although the heterogeneity of this epithelium is well documented, little is known about precursor-progeny relationships between distinct thymic epithelial lineages. Here we characterized a thymic epithelial cell subpopulation identified by the cell surface glycoprotein MTS24. These cells contained epithelial progenitor cells that were competent and

Mark Malin; Georg A. Holländer; Richard Boyd; Jason Gill

2002-01-01

165

Regulation of local immunity by airway epithelial cells  

Microsoft Academic Search

Epithelial cells are the first line of defense against invading microbial pathogens. They are important contributors to innate\\u000a mucosal immunity and generate various and sophisticated anti-microbial defense mechanisms, including the formation of a tight\\u000a barrier and secretion of anti-microbial substances as well as inflammatory mediators. To provide these active defense mechanisms,\\u000a epithelial cells functionally express various pattern-recognition receptors. Toll-like receptors

Anja K. Mayer; Alexander H. Dalpke

2007-01-01

166

Mammary stem cell repertoire: new insights in aging epithelial populations  

Microsoft Academic Search

The proliferative lifespan of mammary stem cells was examined in serially transplanted clonal-dominant epithelial populations. Five successive transplant generations were done. The epithelial cell number in each outgrowth expands ?500-fold in nulliparous hosts and ?10?000-fold in impregnated hosts. Despite this, all resulting mammary outgrowths showed lineal identity with the original. Growth senescence was observed in some implants beginning at the

Gilbert H Smith; Corinne A Boulanger

2002-01-01

167

A new culture medium for human skin epithelial cells  

Microsoft Academic Search

Summary  A new culture medium, NCTC 168, has been designed for human skin epithelial cells. This medium formulation was developed,\\u000a by combining and testing at various concentrations, components of media NCTC 135 and 163, since a 1?1 mixture of these two\\u000a media with 10% horse serum supplement was found to promote epithelial cell outgrowth from human skin explants. The buffer\\u000a system

Floyd M. Price; Richard F. Camalier; Raymond Gantt; William G. Taylor; Gilbert H. Smith; Katherine K. Sanford

1980-01-01

168

Interaction of bacteria and bacterial toxins with intestinal epithelial cells  

Microsoft Academic Search

The epithelium of the intestinal tract is a key barrier between the external environment and the internal body environment.\\u000a Intestinal epithelial cells are targets for luminal bacteria and viruses and must discriminate between pathogenic and nonpathogenic\\u000a commensal organisms. Pathogenic bacteria and their secreted products influence epithelial cell function and induce diarrhea\\u000a by numerous mechanisms that range from an effect on

Asma Nusrat; Shanthi V. Sitaraman; Andrew Neish

2001-01-01

169

Regulated Mucin Secretion from Airway Epithelial Cells  

PubMed Central

Secretory epithelial cells of the proximal airways synthesize and secrete gel-forming polymeric mucins. The secreted mucins adsorb water to form mucus that is propelled by neighboring ciliated cells, providing a mobile barrier which removes inhaled particles and pathogens from the lungs. Several features of the intracellular trafficking of mucins make the airway secretory cell an interesting comparator for the cell biology of regulated exocytosis. Polymeric mucins are exceedingly large molecules (up to 3?×?106?Da per monomer) whose folding and initial polymerization in the ER requires the protein disulfide isomerase Agr2. In the Golgi, mucins further polymerize to form chains and possibly branched networks comprising more than 20 monomers. The large size of mucin polymers imposes constraints on their packaging into transport vesicles along the secretory pathway. Sugar side chains account for >70% of the mass of mucins, and their attachment to the protein core by O-glycosylation occurs in the Golgi. Mature polymeric mucins are stored in large secretory granules ?1??m in diameter. These are translocated to the apical membrane to be positioned for exocytosis by cooperative interactions among myristoylated alanine-rich C kinase substrate, cysteine string protein, heat shock protein 70, and the cytoskeleton. Mucin granules undergo exocytic fusion with the plasma membrane at a low basal rate and a high stimulated rate. Both rates are mediated by a regulated exocytic mechanism as indicated by phenotypes in both basal and stimulated secretion in mice lacking Munc13-2, a sensor of the second messengers calcium and diacylglycerol (DAG). Basal secretion is induced by low levels of activation of P2Y2 purinergic and A3 adenosine receptors by extracellular ATP released in paracrine fashion and its metabolite adenosine. Stimulated secretion is induced by high levels of the same ligands, and possibly by inflammatory mediators as well. Activated receptors are coupled to phospholipase C by Gq, resulting in the generation of DAG and of IP3 that releases calcium from apical ER. Stimulated secretion requires activation of the low affinity calcium sensor Synaptotagmin-2, while a corresponding high affinity calcium sensor in basal secretion is not known. The core exocytic machinery is comprised of the SNARE proteins VAMP8, SNAP23, and an unknown Syntaxin protein, together with the scaffolding protein Munc18b. Common and distinct features of this exocytic system in comparison to neuroendocrine cells and neurons are highlighted.

Adler, Kenneth B.; Tuvim, Michael J.; Dickey, Burton F.

2013-01-01

170

Effect of freezing on lens epithelial cell growth.  

PubMed

The effect of freezing on the growth of rat lens epithelial cells was studied in vitro. We found that 80% of the lens epithelial cells died after freezing at -45 degrees C for two hours and that the surviving cells could grow with the addition of growth factors or when placed on a sheet of type 4 collagen, but not when placed on a plain plastic culture dish. These results suggest that the surviving cells are at the Go phase of the cell cycle and that type 4 collagen or growth factors can initiate cell division. PMID:3294380

Fukaya, Y; Hara, T; Hara, T; Iwata, S

1988-05-01

171

Epimorphin Functions as a Key Morphoregulator for Mammary Epithelial Cells  

PubMed Central

Hepatocyte growth factor (HGF) and EGF have been reported to promote branching morphogenesis of mammary epithelial cells. We now show that it is epimorphin that is primarily responsible for this phenomenon. In vivo, epimorphin was detected in the stromal compartment but not in lumenal epithelial cells of the mammary gland; in culture, however, a subpopulation of mammary epithelial cells produced significant amounts of epimorphin. When epimorphin-expressing epithelial cell clones were cultured in collagen gels they displayed branching morphogenesis in the presence of HGF, EGF, keratinocyte growth factor, or fibroblast growth factor, a process that was inhibited by anti-epimorphin but not anti-HGF antibodies. The branch length, however, was roughly proportional to the ability of the factors to induce growth. Accordingly, epimorphin-negative epithelial cells simply grew in a cluster in response to the growth factors and failed to branch. When recombinant epimorphin was added to these collagen gels, epimorphin-negative cells underwent branching morphogenesis. The mode of action of epimorphin on morphogenesis of the gland, however, was dependent on how it was presented to the mammary cells. If epimorphin was overexpressed in epimorphin-negative epithelial cells under regulation of an inducible promoter or was allowed to coat the surface of each epithelial cell in a nonpolar fashion, the cells formed globular, alveoli-like structures with a large central lumen instead of branching ducts. This process was enhanced also by addition of HGF, EGF, or other growth factors and was inhibited by epimorphin antibodies. These results suggest that epimorphin is the primary morphogen in the mammary gland but that growth factors are necessary to achieve the appropriate cell numbers for the resulting morphogenesis to be visualized.

Hirai, Yohei; Lochter, Andre; Galosy, Sybille; Koshida, Shogo; Niwa, Shinichiro; Bissell, Mina J.

1998-01-01

172

Candida albicans triggers interleukin-8 secretion by oral epithelial cells.  

PubMed

Oropharyngeal candidiasis is a frequent opportunistic infection associated with immunocompromised hosts. Candida albicans is the principal species responsible for this infection. Production of interleukin-8 (IL-8), by oral epithelial cells can be expected to play a major role in the recruitment and activation of professional phagocytes at the infected site. The purpose of this study was to determine whether C. albicans triggers secretion of IL-8 by oral epithelial cells in vitro and investigate mechanisms of host cell-fungal interactions that trigger such responses. Oral epithelial cell lines (SCC4, SCC15, and OKF6/TERT-2) as well as primary gingival epithelial cells were used. Epithelial cells were cocultured with C. albicans, strains SC5314, ATCC28366 or ATCC32077, for 24-48 hr, and supernatants were analyzed for IL-8 content by ELISA. A germination-deficient mutant (efg1/efg1 cph1/cph1), otherwise isogenic to strain SC5314, was used to assess the requirement for germination in triggering IL-8 responses. In order to ascertain whether direct contact of yeast with host cells is required to trigger cytokine production, epithelial cells were separated from yeast using cell culture inserts. To test whether IL-8 secretion is dependent on IL-1alpha activity, epithelial cells were challenged with viable C. albicans in the presence or absence of neutralizing anti-IL-1alpha antibody or IL-1ra, and IL-8 secretion was measured in the supernatants. All cell lines and primary cultures responded to C. albicans with an increase in IL-8 secretion. IL-8 responses were contact-dependent, strain-specific, required yeast viability and germination into hyphae, and were in part autoregulated by IL-1alpha. PMID:12668140

Dongari-Bagtzoglou, A; Kashleva, H

2003-04-01

173

ONCOGENE ALTERNATIONS IN IN VITRO TRANSFORMED RAT TRACHEAL EPITHELIAL CELLS  

EPA Science Inventory

Ten derivations of rat tracheal epithelial (RTE) cells, including normal cells, normal primary cultures, 7 tumorigenic cell lines and 1 non-tumorigenic cell line transformed by treatment with 7,12-dimethylbenz(a)anthracene (DMBA), benzo(a)pyrene (BP) and/or 12-0-tetradecanoylphor...

174

Rabbit uterine epithelial cells: Co-culture with spermatozoa  

SciTech Connect

A primary culture of rabbit uterine epithelial cells was established and their effects on sperm function were examined in vitro. Epithelial cells were isolated from uteri of estrous rabbits and cultured on floating collagen gels in phenol red-free medium supplemented with 5% fetal bovine serum. Light microscopy and keratin staining showed that the epithelial cell population established in culture had morphological characteristics similar to that seen in the intact endometrium. Cells were cultured with {sup 3}H-leucine and uptake of label by cells and its incorporation into cellular and secretory proteins determined. When compared to cells cultured for 24-48 h, incorporation of label into cellular protein was lower at 72-96 h, but secretion increased. Estradiol 17-{beta} did not affect label uptake or incorporation, but did enhance proliferation of cells as judged by total DNA content of the cell population. Analysis of proteins in media by sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography suggested that epithelial and stromal cells synthesis proteins that may be secretory in nature during 72-96 h culture. Twenty-nine to thirty-one h after initiation of epithelial cultures, 1-2 {times} 10{sup 6} sperm were co-incubated with cells and sperm viability, motility, loss of acrosome and fertilizing ability determined.

Boice, M.L.

1988-01-01

175

Diesel exhaust enhances influenza virus infections in respiratory epithelial cells.  

PubMed

Several factors, such as age and nutritional status, can affect the susceptibility to influenza infections. Moreover, exposure to air pollutants, such as diesel exhaust (DE), has been shown to affect respiratory virus infections in rodent models. Influenza virus primarily infects and replicates in respiratory epithelial cells, which are also a major targets for inhaled DE. Using in vitro models of human respiratory epithelial cells, we determined the effects of an aqueous-trapped solution of DE (DE(as)) on influenza infections. Differentiated human nasal and bronchial epithelial cells, as well as A549 cells, were exposed to DE(as) and infected with influenza A/Bangkok/1/79. DE(as) enhanced the susceptibility to influenza virus infection in all cell models and increased the number of influenza-infected cells within 24 h post-infection. This was not caused by suppressing antiviral mediator production, since interferon (IFN) beta levels, IFN-dependent signaling, and IFN-stimulated gene expression were also enhanced by exposure to DE(as). Many of the adverse effects induced by DE exposure are mediated by oxidative stress. Exposure to DE(as) used in these studies generated oxidative stress in respiratory epithelial cells, and addition of the antioxidant glutathione-ethylester (GSH-ET) reversed the effects of DE(as) on influenza infections. Furthermore, DE(as) increased influenza virus attachment to respiratory epithelial cells within 2 h post-infection. Taken together, the results presented here suggest that in human respiratory epithelial cells oxidative stress generated by DE(as) increases the susceptibility to influenza infection and that exposure to DE(as) increases the ability of the virus to attach to and enter respiratory epithelial cells. PMID:15772371

Jaspers, Ilona; Ciencewicki, Jonathan M; Zhang, Wenli; Brighton, Luisa E; Carson, Johnny L; Beck, Melinda A; Madden, Michael C

2005-03-16

176

The Epithelial Cell in Lung Health and Emphysema Pathogenesis  

PubMed Central

Cigarette smoking is the primary cause of the irreversible lung disease emphysema. Historically, inflammatory cells such as macrophages and neutrophils have been studied for their role in emphysema pathology. However, recent studies indicate that the lung epithelium is an active participant in emphysema pathogenesis and plays a critical role in the lung’s response to cigarette smoke. Tobacco smoke increases protease production and alters cytokine expression in isolated epithelial cells, suggesting that these cells respond potently even in the absence of a complete inflammatory program. Tobacco smoke also acts as an immunosuppressant, reducing the defense function of airway epithelial cells and enhancing colonization of the lower airways. Thus, the paradigm that emphysema is strictly an inflammatory-cell based disease is shifting to consider the involvement of resident epithelial cells. Here we review the role of epithelial cells in lung development and emphysema. To better understand tobacco-epithelial interactions we performed microarray analyses of RNA from human airway epithelial cells exposed to smoke extract for 24 hours. These studies identified differential regulation of 425 genes involved in diverse biological processes, such as apoptosis, immune function, cell cycle, signal transduction, proliferation, and antioxidants. Some of these genes, including VEGF, glutathione peroxidase, IL-13 receptor, and cytochrome P450, have been previously reported to be altered in the lungs of smokers. Others, such as pirin, cathepsin L, STAT1, and BMP2, are shown here for the first time to have a potential role in smoke-associated injury. These data broaden our understanding of the importance of epithelial cells in lung health and cigarette smoke-induced emphysema.

Mercer, Becky A.; Lemaitre, Vincent; Powell, Charles A.; D'Armiento, Jeanine

2009-01-01

177

Characteristics and EGFP expression of goat mammary gland epithelial cells.  

PubMed

The aims of this study were (i) to establish a goat mammary gland epithelial (GMGE) cell line, and (ii) to determine if these GMGE cells could be maintained long-term in culture by continuous subculturing following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Primary culture of GMGE cells was achieved by outgrowth of migrating cells from the fragments of the mammary gland tissue of a lactating goat. The passage 16 GMGE cells were transfected with EGFP gene using lipofection. The expression of Cell keratins of epithelial cells in GMGE cells was test by immunofluorescence. ?eta-Casein gene mRNA was test for GMGE cells by RT-PCR. The results showed that when grown at low density on a plastic substratum, the GMGE cells formed islands, and when grown to confluency, the cells formed a monolayer and aggregated with the characteristic cobble-stone morphology of epithelial cells. GMGE cells could form dome-like structure which looked like nipple, and the lumen-like structures formed among the cells. Several blister-like structures appeared in the appearance of the cells. The GMGE cells contained different cell types, majority of the cells were short shuttle-like or polygon which were beehive-like. A part of cells were round and flat, a small number of cells were elongated. Some of the GMGE cells contained milk drops. The cell nuclei were round which had 2-4 obvious cores. The expression of Cell keratins demonstrated the property of epithelial cells in GMGE cells by immunofluorescence. The GMGE cells could express transcript encoding a ?eta-Casein protein. EGFP gene was successfully transferred into the GMGE cells, and the transfected cells could be maintained long-term in culture by continuous subculturing. In conclusion, we have established a EGFP gene transfected GMGE (ET-GMGE) cell line and maintained it long-term in culture by continuous subculturing. PMID:20113446

Zheng, Y-M; He, X-Y; Zhang, Y

2010-12-01

178

Anatomical location and culture of equine corneal epithelial stem cells.  

PubMed

OBJECTIVE: To identify morphologically the locations of equine corneal epithelial stem cells (CESCs) and to culture these cells. ANIMALS STUDIED: We studied the eyes of 12 adult thoroughbred horses. PROCEDURES: Eye tissues were immunostained for two positive stem cell markers (p63, CK14) and one negative marker (CK3) to identify the locations of CESCs, so we could compare their immunostaining patterns with those of human stem cells previously reported. We compared the proliferation rates and morphological features of epithelial cells isolated from the corneal limbus and central cornea. RESULTS: Undifferentiated cells expressing the same immunostaining pattern as human CESCs were present in the equine corneal limbus. Cultured epithelial cells isolated from the limbus expressed the same immunostaining pattern that CESCs show histologically, but cells isolated from the central cornea did not proliferate and could not be evaluated. CONCLUSIONS: Equine CESCs were localized in the epithelial basal layer of the corneal limbus, where melanocytes reside. They could be cultured without loss of their undifferentiated nature. When collecting such stem cells, it may be useful to harvest and culture corneal epithelial tissues in the limbus where melanocytes serve as an indicator of the collecting area. PMID:23710670

Moriyama, Hidekazu; Kasashima, Yoshinori; Kuwano, Atsutoshi; Wada, Shinya

2013-05-28

179

Culture and characterisation of epithelial cells from human pterygia  

PubMed Central

BACKGROUND/AIMS—Pterygia are a common disorder of the ocular surface. The disease represents a chronic fibrovascular and degenerative process thought to originate at the conjunctival-corneal junction, where altered limbal stem cells are proposed to be the cell of origin. Extensive epidemiological evidence exists to implicate ultraviolet B irradiation in the pathogenesis of pterygia. To date no animal or in vitro culture model has been developed to test such an hypothesis. The aim of this study was to establish and characterise a pure population of epithelial cells derived from pterygium tissue.?METHODS—Tissue specimens were obtained from patients undergoing pterygium excision. Explants were cultured in either serum free or serum supplemented medium. Primary and passaged cells were processed for light microscopy, analysed by flow cytometry, and characterised immunohistochemically using specific antibodies.?RESULTS—In serum free culture, cuboidal cells with typical morphology of epithelial cells migrated from the pterygium explants from 3 days onwards and eventually formed a cohesive monolayer. Passaged cells consisted of 98.4% cytokeratin positive cells and demonstrated immunoreactivity for multiple cytokeratins, including AE1, AE3, AE5, but were negative for AE8. These cells also expressed an epithelial specific antigen, together with vimentin and mucin, as did epithelial cells in sections of pterygia.?CONCLUSIONS—A relatively simple method of isolating pterygium epithelial cells has been established. Cultured pterygium epithelial cells are phenotypically and functionally similar to their in vivo counterparts with respect to keratin, vimentin, and mucin expression. In vitro assays using these cells may aid in elucidating the pathogenesis of pterygia.??

Di, G; Tedla, N.; Kumar, R.; McCluskey, P.; Lloyd, A.; Coroneo, M.; Wakefield, D.

1999-01-01

180

Epithelial to Mesenchymal Transition of Mesothelial Cells in Tuberculous Pleurisy  

PubMed Central

Purpose Tuberculous pleurisy is the most frequent extrapulmonary manifestation of tuberculosis. In spite of adequate treatment, pleural fibrosis is a common complication, but the mechanism has not been elucidated. This study is to determine whether epithelial to mesenchymal transition (EMT) of mesothelial cells occurs in tuberculous pleurisy. Materials and Methods Normal pleural mesothelial cells, isolated from irrigation fluids during operations for primary spontaneous pneumothorax, were characterized by immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). These cells were treated in vitro with various cytokines, which were produced in the effluents of tuberculous pleurisy. The isolated cells from the effluents of tuberculous pleurisy were analyzed by immunofluorescence and RT-PCR analysis. Results The isolated cells from the irrigation fluid of primary spontaneous pneumothorax had epithelial characteristics. These cells, with transforming growth factor-?1 and/or interleukin-1? treatment, underwent phenotypic transition from epithelial to mesenchymal cells, with the loss of epithelial morphology and reduction in cytokeratin and E-cadherin expression. Effluent analysis from tuberculous pleurisy using immunofluorescence and RT-PCR demonstrated two phenotypes that showed mesenchymal characteristics and both epithelial & mesencymal characteristics. Conclusion Our results suggest that pleural mesothelial cells in tuberculous pleurisy have been implicated in pleural fibrosis through EMT.

Kim, Changhwan; Park, Sung-Hoon; Hwang, Yong Il; Jang, Seung Hun; Kim, Cheol Hong; Jung, Ki-Suck; Min, Kwangseon; Lee, Jae Woong; Jang, Young Sook

2011-01-01

181

Epithelial cell guidance by self-generated EGF gradients.  

PubMed

Cancer epithelial cells often migrate away from the primary tumor to invade into the surrounding tissues. Their migration is commonly assumed to be directed by pre-existent spatial gradients of chemokines and growth factors in the target tissues. Unexpectedly however, we found that the guided migration of epithelial cells is possible in vitro in the absence of pre-existent chemical gradients. We observed that both normal and cancer epithelial cells can migrate persistently and reach the exit along the shortest path from microscopic mazes filled with uniform concentrations of media. Using microscale engineering techniques and biophysical models, we uncovered a self-guidance strategy during which epithelial cells generate their own guiding cues under conditions of biochemical confinement. The self-guidance strategy depends on the balance between three interdependent processes: epidermal growth factor (EGF) uptake by the cells (U), the restricted transport of EGF through the structured microenvironment (T), and cell chemotaxis toward the resultant EGF gradients (C). The UTC self-guidance strategy can be perturbed by inhibition of signalling through EGF-receptors and appears to be independent from chemokine signalling. Better understanding of the UTC self-guidance strategy could eventually help devise new ways for modulating epithelial cell migration and delaying cancer cell invasion or accelerating wound healing. PMID:22314635

Scherber, Cally; Aranyosi, Alexander J; Kulemann, Birte; Thayer, Sarah P; Toner, Mehmet; Iliopoulos, Othon; Irimia, Daniel

2012-02-08

182

Characterization of a continuous feline mammary epithelial cell line susceptible to feline epitheliotropic viruses  

Microsoft Academic Search

Mucosal epithelial cells are the primary targets for many common viral pathogens of cats. Viral infection of epithelia can damage or disrupt the epithelial barrier that protects underlying tissues. In vitro cell culture systems are an effective means to study how viruses infect and disrupt epithelial barriers, however no true continuous or immortalized feline epithelial cell culture lines are available.

Patricia Pesavento; Hongwei Liu; Robert J. Ossiboff; Karla M. Stucker; Anna Heymer; Lee Millon; Jason Wood; Deborah van der List; John S. L. Parker

2009-01-01

183

Expression of vimentin by rabbit corneal epithelial cells during wound repair  

Microsoft Academic Search

Intermediate filaments of epithelial cells generally consist of specific combinations of keratins. However, cultured epithelial cells from certain tissues and some epithelial tumors have been shown also to express vimentin. In the present study, the expression of vimentin by epithelial cells in healing corneal wounds (partial thickness penetrating wounds) and in tissue culture was analyzed. Both immunohistochemical and immunotransblot analyses

N. SundarRaj; J. D. Rizzo; S. C. Anderson; J. P. Gesiotto

1992-01-01

184

Trans-sialidase Stimulates Eat Me Response from Epithelial Cells  

PubMed Central

Epithelial cell invasion by the protozoan parasite Trypanosoma cruzi is enhanced by the presence of an enzyme expressed on its cell surface during the trypomastigote life cycle stage. The enzyme, trans-sialidase (TS), is a member of one of the largest gene families expressed by the parasite and the role of its activity in mediating epithelial cell entry has not hitherto been understood. Here we show that the T. cruzi TS generates an eat me signal which is capable of enabling epithelial cell entry. We have utilized purified, recombinant, active (TcTS) and inactive (TcTS2V0) TS coated onto beads to challenge an epithelial cell line. We find that TS activity acts upon G protein coupled receptors present at the epithelial cell synapse with the coated bead, thereby enhancing cell entry. By so doing, we provide evidence that TS proteins bind glycans, mediate the formation of distinct synaptic domains and promote macropinocytotic uptake of microparticles into a perinuclear compartment in a manner which may emulate entosis.

Butler, Claire E; de Carvalho, Tecia M U; Grisard, Edmundo C; Field, Robert A; Tyler, Kevin M

2013-01-01

185

Niche regulation of corneal epithelial stem cells at the limbus  

Microsoft Academic Search

Among all adult somatic stem cells, those of the corneal epithelium are unique in their exclusive location in a defined limbal structure termed Palisades of Vogt. As a result, surgical engraftment of limbal epithelial stem cells with or without ex vivo expansion has long been practiced to restore sights in patients inflicted with limbal stem cell deficiency. Nevertheless, compared to

Wei Li; Yasutaka Hayashida; Ying-Ting Chen; Scheffer CG Tseng

2007-01-01

186

Cholera toxin stimulation of human mammary epithelial cells in culture  

SciTech Connect

Addition of cholera toxin to human mammary epithelial cultures derived from reduction mammoplasties and primary carcinomas greatly stimulated cell growth and increased the number of times the cells could be successfully subcultured. Other agents known to increase intracellular cAMP levels were also growth stimulatory. The increased growth potential conferred by cholera toxin enhances the usefulness of this cell culture system.

Stampfer, M.R.

1982-06-01

187

Organization of the cytokeratin network in an epithelial cell  

Microsoft Academic Search

The cytoskeleton is a dynamic three-dimensional structure mainly located in the cytoplasm. It is involved in many cell functions such as mechanical signal transduction and maintenance of cell integrity. Among the three cytoskeletal components, intermediate filaments (the cytokeratin in epithelial cells) are the best candidates for this mechanical role. A model of the establishment of the cytokeratin network of an

Stéphanie Portet; Ovide Arino; Jany Vassy; Damien Schoëvaërt

2003-01-01

188

Menstrual effluent induces epithelial-mesenchymal transitions in mesothelial cells  

Microsoft Academic Search

BACKGROUND: Menstrual effluent affects mesothelial cell (MC) morphology. We evaluated whether these changes were consistent with epithelial-mesenchymal transitions (EMT). METHODS: Monolayer cultures of MC were incubated overnight in conditioned media, prepared from cells isolated form menstrual effluent, with or with- out kinase and ATP inhibitors. Changes in cell morphology were monitored using time-lapse video microscopy and immunohistochemistry. Effects on the

A. Y. Demir; P. G. Groothuis; A. W. Nap; C. Punyadeera; A. F. P. M. de Goeij; J. L. H. Evers; G. A. J. Dunselman

2004-01-01

189

Intestinal Epithelial Cells Secrete Exosome-like Vesicles  

Microsoft Academic Search

Background & Aims: Given the observations that intestinal epithelial cells (IECs) can present antigens to CD4+ T lymphocytes and that professional antigen-presenting cells secrete exosomes (antigen-presenting vesicles), we hypothesized that IECs may secrete exosomes carrying molecules implicated in antigen presentation, which may be able to cross the basement membrane and convey immune information to noncontiguous immune cells. Methods: Human IEC

Guillaume Van Niel; Graça Raposo; Céline Candalh; Muriel Boussac; Robert Hershberg; Nadine Cerf-Bensussan; Martine Heyman

2001-01-01

190

Cytokine Secretion by Cystic Fibrosis Airway Epithelial Cells  

Microsoft Academic Search

It is controversial whether mutations in cystic fibrosis transmembrane conductance regulator intrinsically dysregulate inflammation. We characterized passage 2 human tracheobronchial epithelial cell cul- tures morphologically and physiologically and determined whether cytokine production or nuclear factor-B activation was systemati- cally altered in cystic fibrosis (CF) cells. Non-CF and CF cells originat- ing from a total of 33 and 25 lungs, respectively,

Marie N. Becker; Mariam S. Sauer; Marianne S. Muhlebach; Andrew J. Hirsh; Qi Wu; Margrith W. Verghese; Scott H. Randell

191

ISOLATION AND SERIAL CULTIVATION OF RABBIT SKIN EPITHELIAL CELLS  

Microsoft Academic Search

A method to isolate and to serially cultivate rabbit skin epithelial cells from adult trunk skin has been developed. Using a collagen gel as substrate and trypsin and EDTA to dissociate cells, nonproliferative primary cultures of rabbit cells may be converted to proliferative populations, and at least 3 serial passages achieved. In the presence of large concentrations of methotrexate (up

Su-Chin Liu; Marvin Karasek

1978-01-01

192

Establishment of a corneal epithelial cell line spontaneously derived from human limbal cells  

Microsoft Academic Search

The objective of this study was to establish a spontaneously derived human corneal epithelial cell line from a normal human limbus that retains differentiation potential and proliferative properties under continuous cell culture. After 50 passages of epithelial cells obtained from human limbal tissue a cell line spontaneously emerged. The immortalized cells showed a cobblestone appearance and displayed dense microvilli on

Jingbo liu; Ge Song; Zhichong Wang; Bing Huang; Qianying Gao; Bingqian Liu; Ying Xu; Xuanwei Liang; Ping Ma; Nan Gao; Jian Ge

2007-01-01

193

Detection of ?-defensins secreted by human oral epithelial cells  

Microsoft Academic Search

Human ?-defensins are antimicrobial peptides that may be critical in the innate immune response to infection. hBD1 and hBD2 are expressed in oral epithelial cells and are detected near the surface of oral tissue, consistent with a role in the epithelial protective barrier function. In this report, we examine secretion of ?-defensins in vitro and in biological fluid using ProteinChip®

Deborah L Diamond; Janet R Kimball; Suttichai Krisanaprakornkit; Tomas Ganz; Beverly A Dale

2001-01-01

194

Ozone exposed epithelial cells modify cocultured natural killer cells.  

PubMed

Ozone (O3) causes significant adverse health effects worldwide. Nasal epithelial cells (NECs) are among the first sites within the respiratory system to be exposed to inhaled air pollutants. They recruit, activate, and interact with immune cells via soluble mediators and direct cell-cell contacts. Based on our recent observation demonstrating the presence of natural killer (NK) cells in nasal lavages, the goal of this study was to establish a coculture model of NECs and NK cells and examine how exposure to O3 modifies this interaction. Flow cytometry analysis was used to assess immunophenotypes of NK cells cocultured with either air- or O3-exposed NECs. Our data show that coculturing NK cells with O3-exposed NECs decreased intracellular interferon-? (IFN-?), enhanced, albeit not statistically significant, IL-4, and increased CD16 expression on NK cells compared with air controls. Additionally, the cytotoxicity potential of NK cells was reduced after coculturing with O3-exposed NECs. To determine whether soluble mediators released by O3-exposed NECs caused this shift, apical and basolateral supernatants of air- and O3-exposed NECs were used to stimulate NK cells. While the conditioned media of O3-exposed NECs alone did not reduce intracellular IFN-?, O3 enhanced the expression of NK cell ligands ULBP3 and MICA/B on NECs. Blocking ULBP3 and MICA/B reversed the effects of O3-exposed NECs on IFN-? production in NK cells. Taken together, these data showed that interactions between NECs and NK cells in the context of O3 exposure changes NK cell activity via direct cell-cell interactions and is dependent on ULBP3/MICA/B expressed on NECs. PMID:23241529

Müller, Loretta; Brighton, Luisa E; Jaspers, Ilona

2012-12-14

195

Change in cell shape is required for matrix metalloproteinase-induced epithelial-mesenchymal transition of mammary epithelial cells  

SciTech Connect

Cell morphology dictates response to a wide variety of stimuli, controlling cell metabolism, differentiation, proliferation, and death. Epithelial-mesenchymal transition (EMT) is a developmental process in which epithelial cells acquire migratory characteristics, and in the process convert from a 'cuboidal' epithelial structure into an elongated mesenchymal shape. We had shown previously that matrix metalloproteinase-3 (MMP3) can stimulate EMT of cultured mouse mammary epithelial cells through a process that involves increased expression of Rac1b, a protein that stimulates alterations in cytoskeletal structure. We show here that cells treated with MMP-3 or induced to express Rac1b spread to cover a larger surface, and that this induction of cell spreading is a requirement of MMP-3/Rac1b-induced EMT. We find that limiting cell spreading, either by increasing cell density or by culturing cells on precisely defined micropatterned substrata, blocks expression of characteristic markers of EMT in cells treated with MMP-3. These effects are not caused by general disruptions in cell signaling pathways, as TGF-{beta}-induced EMT is not affected by similar limitations on cell spreading. Our data reveal a previously unanticipated cell shape-dependent mechanism that controls this key phenotypic alteration and provide insight into the distinct mechanisms activated by different EMT-inducing agents.

Nelson, Celeste M.; Khauv, Davitte; Bissell, Mina J.; Radisky, Derek C.

2008-06-26

196

Reactive oxygen molecule-mediated injury in endothelial and renal tubular epithelial cells in vitro  

Microsoft Academic Search

Reactive oxygen molecule-mediated injury in endothelial and renal tubular epithelial cells in vitro. To investigate renal tubular epithelial cell injury mediated by reactive oxygen molecules and to explore the relative susceptibility of epithelial cells and endothelial cells to oxidant injury, we determined cell injury in human umbilical vein endothelial cells and in four renal tubular epithelial cell lines including LLC-PK1,

Sharon P Andreoli; James A McAteer; Coleen Mallett

1990-01-01

197

Epithelial Cancer Cell Migration: A Role for Chemokine Receptors?  

Microsoft Academic Search

We investigated the possibility that chemokine gradients influence migration of human ovarian epithelial tumor cells. Of 14 chemokine receptors investigated, only CXCR4 was expressed on ovarian cancer cells. CXCR4 mRNA localized to a subpopulation of tumor cells in ovarian cancer biopsies. Ovarian cancer cell lines and cells freshly isolated from ascites expressed CXCR4 protein. The CXCR4 ligand, CXCL12, was found

Chris J. Scotton; Julia L. Wilson; David Milliken; Gordon Stamp; Frances R. Balkwill

2001-01-01

198

Epithelial cells and macrophages in myasthenia gravis thymus culture.  

PubMed

Monolayer culture of thymic nonlymphoid cells derived from female patients with myasthenia gravis (MG) and individuals who underwent heart surgery was established to investigate the cellular composition of the thymic microenvironment and the interaction of nonlymphoid cells with autologous thymocytes. Thymic epithelial cells were identified by immunoperoxidase staining using monoclonal antibodies (mAbs) specific for cytokeratin and MR6 and MR19 antigens expressed on cortical and medullary epithelial cells, respectively. Macrophages were characterized by determination of alpha-naphthyl acetate esterase activity and detection of M1 antigen by mAb. It was demonstrated that in MG thymus cultures the number of cortical MR6+ epithelial cells is significantly reduced, and the ability of the remaining MR6+ cells to bind autologous thymocytes is markedly affected. On the other hand, the number of macrophages and the interaction of those cells with thymocytes were similar in MG and control thymus cultures. Since MR6+ epithelial cells are numerically and functionally affected in MG, maturational events of T cells occurring in the inner cortex may be altered. The mechanisms underlying the induction and expansion of T helper clones in MG are discussed. PMID:2390810

Popeskovic, L; Apostolski, S; Isakovic, K

1990-09-01

199

In vitro interaction of Mycobacterium avium with intestinal epithelial cells.  

PubMed Central

Human intestinal epithelial cell monolayers were inoculated with cultures of Mycobacterium avium serotype 2, 8, or 10 that were viable, autoclaved, Formalin killed, exposed to UV light, or suspended in anti-M. avium serotype 2 serum. The effects of four reagents known to block phagocytosis or endocytosis (cytochalasin B, dibutyryl cyclic adenosine monophosphate, iodoacetate, and 2,4-dinitrophenol) on the bacteria-cell interaction were also studied. The maximum uptake of pathogenic M. avium by human intestinal epithelial cells occurred after 2 to 3 h of incubation. Serotype 2 was taken up in greater quantity than serotype 8 or 10. Saprophytic mycobacteria did not attach to or penetrate the host cells. The data showed that viable mycobacteria are ingested by host cells, whereas dead organisms are not. Components of the bacterial cells are partially, but not solely, responsible for the phagocytosis of M. avium serotype 2 by human intestinal epithelial cells. Furthermore, uptake of M. avium by human intestinal epithelial cells was suppressed by reagents which inhibit uptake by known phagocytic cells, suggesting that the mechanism of uptake is an endocytic process induced by virulent mycobacteria.

Mapother, M E; Songer, J G

1984-01-01

200

Proliferating cells in HIV and pamidronate-associated collapsing focal segmental glomerulosclerosis are parietal epithelial cells  

Microsoft Academic Search

Collapsing focal segmental glomerulosclerosis (cFSGS) is characterized by hyperplasia of glomerular epithelial cells. In a mouse model of FSGS and in a patient with recurrent idiopathic FSGS, we identified the proliferating cells as parietal epithelial cells (PECs). In the present study, we have evaluated the origin of the proliferating cells in cFSGS associated with human immunodeficiency virus (HIV) and pamidronate.

H. B. P. M. Dijkman; J. J. Weening; B. Smeets; K. C. N. Verrijp; T H van Kuppevelt; K. K. J. M. Assmann; E. J. Steenbergen; J. F. M. Wetzels; HBPM Dijkman

2006-01-01

201

Cell-adhesion molecule uvomorulin is localized in the intermediate junctions of adult intestinal epithelial cells  

Microsoft Academic Search

Uvomorulin is a cell-adhesion molecule implicated in the compaction process of mouse preimplantation embryos and the aggregation of embryonal carcinoma cells. A rabbit antiserum against purified uvomorulin also reacts with epithelial cells of various adult tissues. In this study, we investigated the localization of uvomorulin on adult intestinal epithelial cells using electron micro- scopic analyses. Uvomorulin was shown to exhibit

K. Boller; D. VESTWEBER; R. KEMLER

1985-01-01

202

Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification.  

PubMed

A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the production of new mouse models of disease and investigations into the hormonal regulation of tissue degeneration (or regeneration) following pathological challenge. The murine reproductive (or estrous) cycle is divided into 4 stages: proestrus, estrus, metestrus, and diestrus. Defined fluctuations in circulating levels of the ovarian steroids 17-?-estradiol and progesterone, the gonadotropins luteinizing and follicle stimulating hormones, and the luteotropic hormone prolactin signal transition through these reproductive stages. Changes in cell typology within the murine vaginal canal reflect these underlying endocrine events. Daily assessment of the relative ratio of nucleated epithelial cells, cornified squamous epithelial cells, and leukocytes present in vaginal smears can be used to identify murine estrous stages. The degree of invasiveness, however, employed in collecting these samples can alter reproductive status and elicit an inflammatory response that can confound cytological assessment of smears. Here, we describe a simple, non-invasive protocol that can be used to determine the stage of the estrous cycle of a female mouse without altering her reproductive cycle. We detail how to differentiate between the four stages of the estrous cycle by collection and analysis of predominant cell typology in vaginal smears and we show how these changes can be interpreted with respect to endocrine status. PMID:23007862

McLean, Ashleigh C; Valenzuela, Nicolas; Fai, Stephen; Bennett, Steffany A L

2012-09-15

203

Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification  

PubMed Central

A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the production of new mouse models of disease and investigations into the hormonal regulation of tissue degeneration (or regeneration) following pathological challenge. The murine reproductive (or estrous) cycle is divided into 4 stages: proestrus, estrus, metestrus, and diestrus. Defined fluctuations in circulating levels of the ovarian steroids 17-?-estradiol and progesterone, the gonadotropins luteinizing and follicle stimulating hormones, and the luteotropic hormone prolactin signal transition through these reproductive stages. Changes in cell typology within the murine vaginal canal reflect these underlying endocrine events. Daily assessment of the relative ratio of nucleated epithelial cells, cornified squamous epithelial cells, and leukocytes present in vaginal smears can be used to identify murine estrous stages. The degree of invasiveness, however, employed in collecting these samples can alter reproductive status and elicit an inflammatory response that can confound cytological assessment of smears. Here, we describe a simple, non-invasive protocol that can be used to determine the stage of the estrous cycle of a female mouse without altering her reproductive cycle. We detail how to differentiate between the four stages of the estrous cycle by collection and analysis of predominant cell typology in vaginal smears and we show how these changes can be interpreted with respect to endocrine status.

McLean, Ashleigh C.; Valenzuela, Nicolas; Fai, Stephen; Bennett, Steffany A.L.

2012-01-01

204

Extracellular matrix proteins regulate epithelial-mesenchymal transition in mammary epithelial cells.  

PubMed

Mouse mammary epithelial cells undergo transdifferentiation via epithelial-mesenchymal transition (EMT) upon treatment with matrix metalloproteinase-3 (MMP3). In rigid microenvironments, MMP3 upregulates expression of Rac1b, which translocates to the cell membrane to promote induction of reactive oxygen species and EMT. Here we examine the role of the extracellular matrix (ECM) in this process. Our data show that the basement membrane protein laminin suppresses the EMT response in MMP3-treated cells, whereas fibronectin promotes EMT. These ECM proteins regulate EMT via interactions with their specific integrin receptors. ?6-integrin sequesters Rac1b from the membrane and is required for inhibition of EMT by laminin. In contrast, ?5-integrin maintains Rac1b at the membrane and is required for the promotion of EMT by fibronectin. Understanding the regulatory role of the ECM will provide insight into mechanisms underlying normal and pathological development of the mammary gland. PMID:23660532

Chen, Qike K; Lee, Kangae; Radisky, Derek C; Nelson, Celeste M

2013-05-06

205

Genes involved in TGF?1-driven epithelial-mesenchymal transition of renal epithelial cells are topologically related in the human interactome map  

Microsoft Academic Search

BACKGROUND: Understanding how mesenchymal cells arise from epithelial cells could have a strong impact in unveiling mechanisms of epithelial cell plasticity underlying kidney regeneration and repair. In primary human tubular epithelial cells (HUTEC) under different TGF?1 concentrations we had observed epithelial-to-mesenchymal transition (EMT) but not epithelial-myofibroblast transdifferentiation. We hypothesized that the process triggered by TGF?1 could be a dedifferentiation event.

Stefano Campanaro; Simone Picelli; Rossella Torregrossa; Laura Colluto; Monica Ceol; Dorella Del Prete; Angela D'Angelo; Giorgio Valle; Franca Anglani

2007-01-01

206

Role of Epithelial-Mesenchymal Transition in the Formation of Normal and Neoplastic Mammary Epithelial Stem Cells.  

National Technical Information Service (NTIS)

The epithelial-mesenchymal transition (EMT) has been found by us and others to induce normal and neoplastic mammary epithelial cells (MECs) to acquire mesenchymal traits and, in addition, many of the characteristics of stem cells. However, none of these o...

Z. Keckesova

2011-01-01

207

Purification and unique properties of mammary epithelial stem cells  

Microsoft Academic Search

Elucidation of the cellular and molecular mechanisms that maintain mammary epithelial tissue integrity is of broad interest and paramount to the design of more effective treatments for breast cancer. Evidence from both in vitro and in vivo experiments suggests that mammary cell differentiation is a hierarchical process originating in an uncommitted stem cell with self-renewal potential. However, analysis of the

John Stingl; Peter Eirew; Ian Ricketson; Mark Shackleton; François Vaillant; David Choi; Haiyan I. Li; Connie J. Eaves

2006-01-01

208

Nipah virus entry and egress from polarized epithelial cells.  

PubMed

Highly pathogenic Nipah virus (NiV) infections are transmitted via airway secretions and urine, commonly via the respiratory route. Epithelial surfaces represent important replication sites in both primary and systemic infection phases. NiV entry and spread from polarized epithelial cells therefore determine virus entry and dissemination within a new host and influence virus shedding via mucosal surfaces in the respiratory and urinary tract. To date, there is no knowledge regarding the entry and exit sites of NiV in polarized epithelial cells. In this report, we show for the first time that NiV can infect polarized kidney epithelial cells (MDCK) from both cell surfaces, while virus release is primarily restricted to the apical plasma membrane. Substantial amounts of basolateral infectivity were detected only after infection with high virus doses, at time points when the integrity of the cell monolayer was largely disrupted as a result of cell-to-cell fusion. Confocal immunofluorescence analyses of envelope protein distribution at early and late infection stages suggested that apical virus budding is determined by the polarized sorting of the NiV matrix protein, M. Studies with stably M-expressing and with monensin-treated cells furthermore demonstrated that M protein transport is independent from the glycoproteins, implying that the M protein possesses an intrinsic apical targeting signal. PMID:23283941

Lamp, Boris; Dietzel, Erik; Kolesnikova, Larissa; Sauerhering, Lucie; Erbar, Stephanie; Weingartl, Hana; Maisner, Andrea

2013-01-02

209

Vectorial secretion of proteoglycans by polarized rat uterine epithelial cells  

Microsoft Academic Search

We have studied proteoglycan secretion using a recently developed system for the preparing of polarized primary cultures of rat uterine epithelial cells. To mimic their native environment better and provide a system for discriminating apical from baso- lateral compartments, we cultured cells on semiperme- able supports impregnated with biomatrix. Keratan sulfate proteoglycans (KSPG) as well as heparan sul- fate-containing molecules

Daniel D. Carson; Jy-Ping Tang; Joanne Julian; Stanley R. Glasser

1988-01-01

210

Human respiratory epithelial cell culture for drug delivery applications  

Microsoft Academic Search

Recent developments in delivering drugs to the lung are driving the need for in vitro methods to evaluate the fate of inhaled medicines. Constraints on experimentation using animals have promoted the use of human respiratory epithelial cell cultures to model the absorption barrier of the lung; with two airway cell lines, 16HBE14o- and Calu-3, and primary cultured human alveolar type

Ben Forbes; Carsten Ehrhardt

2005-01-01

211

Biosystem for the culture and characterisation of epithelial cell tissues  

Microsoft Academic Search

We have designed and realised a new type of microsystem for the electrical characterisation of epithelial cell layers for biomedical diagnostic purposes. We have used deep plasma and other micromachining procedures for the fabrication of vias and microfluidic channels in silicon and glass substrates. Miniaturised cell culture devices have been realised by gluing nano-porous polycarbonate membranes in between two structured

S. Hediger; J. Fontannaz; A. Sayah; W. Hunziker; M. A. M. Gijs

2000-01-01

212

Identification of Phosphorylation Sites on Extracellular Corneal Epithelial Cell Maspin  

PubMed Central

Maspin, a 42-kDa non classical serine protease inhibitor (serpin) is expressed by epithelial cells of various tissues including the cornea. The protein localizes to the nucleus and cytosol, and is present in the extracellular space. While extracellular maspin regulates corneal stromal fibroblast adhesion and inhibits angiogenesis during wound healing in the cornea, the molecular mechanism of its extracellular functions is unclear. We hypothesized that identifying post-translational modifications of maspin, such as phosphorylation, may help decipher its mode of action. The focus of this study was on the identification of phosphorylation sites on extracellular maspin, since the extracellular form of the molecule is implicated in several functions. Multi-stage fragmentation mass spectrometry was used to identify sites of phosphorylation on extracellular corneal epithelial cell maspin. A total of eight serine and threonine phosphorylation sites (Thr50, Ser97, Thr118, Thr157, Ser240, Ser298, Thr310, Ser316) were identified on the extracellular forms of the molecule. Phosphorylation of tyrosine residues on extracellular maspin was not detected on extracellular maspin from corneal epithelial cell, in contrast to breast epithelial cells. This study provides the basis for further investigation into the functional role of phosphorylation of corneal epithelial maspin.

Narayan, Malathi; Mirza, Shama P.; Twining, Sally S.

2011-01-01

213

Growth regulation in Hydra: Relationship between epithelial cell cycle length and growth rate*1  

Microsoft Academic Search

The relationship between epithelial cell production and growth rate was investigated in Hydra attenwcta under different feeding regimes. The increase of epithelial cell number was compared to the duration of the epithelial cell cycle using standard methods of cell cycle analysis. The results indicate that cell cycle changes accompanying changes in feeding regime are not sufficient to explain the altered

THOMAS C. G. BOSCH; CHARLES N. DAVID

1984-01-01

214

Phenotypic characterization of human corneal epithelial cells expanded ex vivo from limbal explant and single cell cultures  

Microsoft Academic Search

Cultivated human corneal epithelial cells have been successfully used for corneal reconstruction. Explant and single cell systems are currently used for human corneal epithelial cultivation. This study was conducted to characterize the phenotypes of human corneal epithelial cells expanded ex vivo by these two culture systems with regard to their growth potential, morphology and antigen expression patterns. Human corneal epithelial

Hyun-Seung Kim; Xiu Jun Song; Cintia S de Paiva; Zhuo Chen; Stephen C Pflugfelder; De-Quan Li

2004-01-01

215

Equine bronchial epithelial cells differentiate into ciliated and mucus producing cells in vitro  

Microsoft Academic Search

We describe a method for creating differentiated equine bronchial epithelial cell cultures that can be used for in vitro studies\\u000a including airway disease mechanisms and pathogen–host interactions. Our method is based on the culturing of human tracheobronchial\\u000a epithelial cells at an air–liquid interface (ALI) in specific serum-free, hormone-supplemented medium. Bronchial epithelial\\u000a cells are isolated and grown on T-Clear® insert membranes.

Ute E. Schwab; M. Leslie Fulcher; Scott H. Randell; M. Julia Flaminio; David G. Russell

2010-01-01

216

Early Response of Mucosal Epithelial Cells During Toxoplasma gondii Infection  

PubMed Central

The innate immune response of mucosal epithelial cells during pathogen invasion plays a central role in immune regulation in the gut. Toxoplasma gondii (T. gondii) is a protozoan intracellular parasite that is usually transmitted through oral infection. Although much of the information on immunity to T. gondii has come from intraperitoneal infection models, more recent studies have revealed the importance of studying immunity following infection through the natural per-oral route. Oral infection studies have identified many of the key players in the intestinal response; however, they have relied on responses detected days to weeks following infection. Much less is known about how the gut epithelial layer senses and reacts during initial contact with the pathogen. Given the importance of epithelial cells during pathogen invasion, this study uses an in vitro approach to isolate the key players and examine the early response of intestinal epithelial cells during infection by T. gondii. We show that human intestinal epithelial cells infected with T. gondii elicit rapid MAPK phosphorylation, NF-?B nuclear translocation, and secretion of interleukin (IL)-8. Both ERK1/2 activation and IL-8 secretion responses were shown to be MyD88 dependent and TLR2 was identified to be involved in the recognition of the parasite regardless of the parasite genotype. Furthermore, we were able to identify additional T. gondii-regulated genes in the infected cells using a pathway-focused array. Together, our findings suggest that intestinal epithelial cells were able to recognize T. gondii during infection, and the outcome is important for modulating intestinal immune responses.

Ju, Chia-Hsin; Chockalingam, Annapoorani; Leifer, Cynthia A.

2012-01-01

217

Modeling epithelial cell homeostasis: Assessing recovery and control mechanisms  

Microsoft Academic Search

Critical to epithelial cell viability is prompt and direct recovery, following a perturbation of cellular conditions. Although\\u000a a number of transporters are known to be activated by changes in cell volume, cell pH, or cell membrane potential, their importance\\u000a to cellular homeostasis has been difficult to establish. Moreover, the coordination among such regulated transporters to enhance\\u000a recovery has received no

Alan M. Weinstein

2004-01-01

218

Infection of Primary Human Bronchial Epithelial Cells by Haemophilus influenzae: Macropinocytosis as a Mechanism of Airway Epithelial Cell Entry  

Microsoft Academic Search

Nontypeable Haemophilus influenzae is an exclusive human pathogen which infects the respiratory epithe- lium. We have initiated studies to explore the interaction of the nontypeable H. influenzae strain 2019 with primary human airway epithelial cells by electron and confocal microscopy. Primary human airway cell cultures were established as monolayers on glass collagen-coated coverslips or on semipermeable membranes at an air-fluid

MARGARET R. KETTERER; JIAN Q. SHAO; DOUGLAS B. HORNICK; BEN BUSCHER; VENKATA K. BANDI; MICHAEL A. APICELLA

1999-01-01

219

Cytolytic vaginosis: misdiagnosed as candidal vaginitis.  

PubMed Central

OBJECTIVES: In this study, 210 women with vaginal discharge and other symptoms/signs of genital pathology suggestive of vulvovaginal candidiasis (VVC) were involved in order to distinguish true WC and cytolytic vaginosis (CV) cases. METHODS: Fungal cultures, 10% potassium hydroxide (KOH) and Gram stained preparations and pH measurements were performed on the vaginal discharge material of each patient. RESULTS: Fifteen patients (7.1%) were diagnosed with cytolytic vaginosis according to their clinical and microbiological findings, including abundant lactobacilli, fragmented epithelial cells and/or free nuclei due to cytolysis, seen in their discharge materials on microscopic examination, but no fungal growth. CONCLUSIONS: The results of this study may contribute to the reports in the literature indicating the importance of such disorders, which are generally misdiagnosed as candidiasis.

Cerikcioglu, Nilgun; Beksac, M Sinan

2004-01-01

220

Epithelial neoplasia in Drosophila entails switch to primitive cell states  

PubMed Central

Only select cell types in an organ display neoplasia when targeted oncogenically. How developmental lineage hierarchies of these cells prefigure their neoplastic propensities is not yet well-understood. Here we show that neoplastic Drosophila epithelial cells reverse their developmental commitments and switch to primitive cell states. In a context of alleviated tissue surveillance, for example, loss of Lethal giant larvae (Lgl) tumor suppressor in the wing primordium induced epithelial neoplasia in its Homothorax (Hth)-expressing proximal domain. Transcriptional profile of proximally transformed mosaic wing epithelium and functional tests revealed tumor cooperation by multiple signaling pathways. In contrast, lgl? clones in the Vestigial (Vg)-expressing distal wing epithelium were eliminated by cell death. Distal lgl? clones, however, could transform when both tissue surveillance and cell death were compromised genetically and, alternatively, when the transcription cofactor of Hippo signaling pathway, Yorkie (Yki), was activated, or when Ras/EGFR signaling was up-regulated. Furthermore, transforming distal lgl? clones displayed loss of Vg, suggesting reversal of their terminal cell fate commitment. In contrast, reinforcing a distal (wing) cell fate commitment in lgl? clones by gaining Vg arrested their neoplasia and induced cell death. We also show that neoplasia in both distal and proximal lgl? clones could progress in the absence of Hth, revealing Hth-independent wing epithelial neoplasia. Likewise, neoplasia in the eye primordium resulted in loss of Elav, a retinal cell marker; these, however, switched to an Hth-dependent primitive cell state. These results suggest a general characteristic of “cells-of-origin” in epithelial cancers, namely their propensity for switch to primitive cell states.

Khan, Sumbul J.; Bajpai, Anjali; Alam, Mohammad Atif; Gupta, Ram P.; Harsh, Sneh; Pandey, Ravi K.; Goel-Bhattacharya, Surbhi; Nigam, Aditi; Mishra, Arati; Sinha, Pradip

2013-01-01

221

Epithelial neoplasia in Drosophila entails switch to primitive cell states.  

PubMed

Only select cell types in an organ display neoplasia when targeted oncogenically. How developmental lineage hierarchies of these cells prefigure their neoplastic propensities is not yet well-understood. Here we show that neoplastic Drosophila epithelial cells reverse their developmental commitments and switch to primitive cell states. In a context of alleviated tissue surveillance, for example, loss of Lethal giant larvae (Lgl) tumor suppressor in the wing primordium induced epithelial neoplasia in its Homothorax (Hth)-expressing proximal domain. Transcriptional profile of proximally transformed mosaic wing epithelium and functional tests revealed tumor cooperation by multiple signaling pathways. In contrast, lgl(-) clones in the Vestigial (Vg)-expressing distal wing epithelium were eliminated by cell death. Distal lgl(-) clones, however, could transform when both tissue surveillance and cell death were compromised genetically and, alternatively, when the transcription cofactor of Hippo signaling pathway, Yorkie (Yki), was activated, or when Ras/EGFR signaling was up-regulated. Furthermore, transforming distal lgl(-) clones displayed loss of Vg, suggesting reversal of their terminal cell fate commitment. In contrast, reinforcing a distal (wing) cell fate commitment in lgl(-) clones by gaining Vg arrested their neoplasia and induced cell death. We also show that neoplasia in both distal and proximal lgl(-) clones could progress in the absence of Hth, revealing Hth-independent wing epithelial neoplasia. Likewise, neoplasia in the eye primordium resulted in loss of Elav, a retinal cell marker; these, however, switched to an Hth-dependent primitive cell state. These results suggest a general characteristic of "cells-of-origin" in epithelial cancers, namely their propensity for switch to primitive cell states. PMID:23708122

Khan, Sumbul J; Bajpai, Anjali; Alam, Mohammad Atif; Gupta, Ram P; Harsh, Sneh; Pandey, Ravi K; Goel-Bhattacharya, Surbhi; Nigam, Aditi; Mishra, Arati; Sinha, Pradip

2013-05-24

222

Respiratory epithelial cell lines exposed to anoxia produced inflammatory mediator  

PubMed Central

Human epithelial cell lines were utilized to examine the effects of anoxia on cellular growth and metabolism. Three normal human epithelial cells lines (A549, NHBE, and BEAS-2B) as well as a cystic fibrosis cell line (IB3-1) and its mutation corrected cell line (C38) were grown in the presence and absence of oxygen for varying periods of time. Interleukin-8 (IL-8) levels were measured by enzyme-linked immunosorbent assay technique. Cellular metabolism and proliferation were assayed by determining mitochondrial oxidative burst activity by tetrazolium compound reduction. The viability of cells was indirectly measured by lactate dehydrogenase release. A549, NHBE, and BEAS-2B cells cultured in the absence of oxygen showed a progressive decrease in metabolic activity and cell proliferation after one to three days. There was a concomitant increase in IL-8 production. Cell lines from cystic fibrosis (CF) patients did not show a similar detrimental effect of anoxia. However, the IL-8 level was significantly increased only in IB3-1 cells exposed to anoxia after two days. Anoxia appears to affect certain airway epithelial cell lines uniquely with decreased cellular proliferation and a concomitant increased production of a cytokine with neutrophilic chemotactic activity. The increased ability of the CF cell line to respond to anoxia with increased secretion of inflammatory cytokines may contribute to the inflammatory damage seen in CF bronchial airway. This study indicates the need to use different cell lines in in vitro studies investigating the role of epithelial cells in airway inflammation and the effects of environmental influences.

Shahriary, Cyrus M.; Nussbaum, Eliezer

2012-01-01

223

Airway epithelial cell response to human metapneumovirus infection  

SciTech Connect

Human metapneumovirus (hMPV) is a major cause of lower respiratory tract infections (LRTIs) in infants, elderly and immunocompromised patients. In this study, we show that hMPV can infect in a similar manner epithelial cells representative of different tracts of the airways. hMPV-induced expression of chemokines IL-8 and RANTES in primary small alveolar epithelial cells (SAE) and in a human alveolar type II-like epithelial cell line (A549) was similar, suggesting that A549 cells can be used as a model to study lower airway epithelial cell responses to hMPV infection. A549 secreted a variety of CXC and CC chemokines, cytokines and type I interferons, following hMPV infection. hMPV was also a strong inducer of transcription factors belonging to nuclear factor (NF)-{kappa}B, interferon regulatory factors (IRFs) and signal transducers and activators of transcription (STATs) families, which are known to orchestrate the expression of inflammatory and immunomodulatory mediators.

Bao, X.; Liu, T.; Spetch, L.; Kolli, D. [Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas (United States); Garofalo, R.P. [Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas (United States); Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas (United States); Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas (United States); Casola, A. [Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas (United States); Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas (United States); Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas (United States)], E-mail: ancasola@utmb.edu

2007-11-10

224

Vaginal Smears and the OEstrous Cycle of the Cat and the Lioness  

Microsoft Academic Search

A study of vaginal smears taken from ten cats during a period of a year showed that their oestrous cycle consists of the following four chief phases: (1) Pro-oeslrus.Characterized by the presence of very numerous nucleated epithelial cells, of fairly uniform shape.

H. Liche; Kazimierz Wodzicki

1939-01-01

225

The Epithelial-to-Mesenchymal Transition and Cancer Stem Cells  

Microsoft Academic Search

\\u000a The epithelial-to-mesenchymal transition (EMT) is a developmental process which is reactivated during carcinoma progression,\\u000a providing tumor cells with enhanced migratory properties, the capacity to invade the stroma, and the ability to metastasize.\\u000a Tumor cells undergoing EMT also acquire stem cell characteristics, suggesting that there is crosstalk between pathways promoting\\u000a EMT and self-renewal, and that the EMT process contributes to the

Jonas Fuxe

226

Protection of human colon epithelial cells against deoxycholate by rottlerin  

Microsoft Academic Search

The bile salt, deoxycholate (DOC), can harm cells and cause disease. Hence, there is interest in identifying compounds capable\\u000a of protecting cells against DOC. In HCT-116 colon epithelial cells, DOC increased generation of reactive oxygen species and\\u000a caused DNA damage and apoptosis. These effects of DOC were inhibited by rottlerin, which is a phenolic compound of plant origin.\\u000a In elucidating

Jennifer M. Longpre; George Loo

2008-01-01

227

Epithelial mesenchymal transition traits in human breast cancer cell lines  

Microsoft Academic Search

Epithelial mesenchymal transition (EMT) has long been associated with breast cancer cell invasiveness and evidence of EMT\\u000a processes in clinical samples is growing rapidly. Genome-wide transcriptional profiling of increasingly larger numbers of\\u000a human breast cancer (HBC) cell lines have confirmed the existence of a subgroup of cell lines (termed Basal B\\/Mesenchymal)\\u000a with enhanced invasive properties and a predominantly mesenchymal gene

T. Blick; E. Widodo; H. Hugo; M. Waltham; M. E. Lenburg; R. M. Neve; E. W. Thompson

2008-01-01

228

Human Alveolar Epithelial Cell Injury Induced by Cigarette Smoke  

Microsoft Academic Search

BackgroundCigarette smoke (CS) is a highly complex mixture and many of its components are known carcinogens, mutagens, and other toxic substances. CS induces oxidative stress and cell death, and this cell toxicity plays a key role in the pathogenesis of several pulmonary diseases.Methodology\\/Principal FindingsWe studied the effect of cigarette smoke extract (CSE) in human alveolar epithelial type I-like (ATI-like) cells.

Beata Kosmider; Elise M. Messier; Hong Wei Chu; Robert J. Mason

2011-01-01

229

Acid Production by Vaginal Flora In Vitro Is Consistent with the Rate and Extent of Vaginal Acidification  

PubMed Central

Perinatally, and between menarche and menopause, increased levels of estrogen cause large amounts of glycogen to be deposited in the vaginal epithelium. During these times, the anaerobic metabolism of the glycogen, by the epithelial cells themselves and/or by vaginal flora, causes the vagina to become acidic (pH ?4). This study was designed to test whether the characteristics of acid production by vaginal flora in vitro can account for vaginal acidity. Eight vaginal Lactobacillus isolates from four species—L. gasseri, L. vaginalis, L. crispatus, and L. jensenii—acidified their growth medium to an asymptotic pH (3.2 to 4.8) that matches the range seen in the Lactobacillus-dominated human vagina (pH 3.6 to 4.5 in most women) (B. Andersch, L. Forssman, K. Lincoln, and P. Torstensson, Gynecol. Obstet. Investig. 21:19–25, 1986; L. Cohen, Br. J. Vener. Dis. 45:241–246, 1969; J. Paavonen, Scand. J. Infect. Dis. Suppl. 40:31–35, 1983; C. Tevi-Bénissan, L. Bélec, M. Lévy, V. Schneider-Fauveau, A. Si Mohamed, M.-C. Hallouin, M. Matta, and G. Grésenguet, Clin. Diagn. Lab. Immunol. 4:367–374, 1997). During exponential growth, all of these Lactobacillus species acidified their growth medium at rates on the order of 106 protons/bacterium/s. Such rates, combined with an estimate of the total number of lactobacilli in the vagina, suggest that vaginal lactobacilli could reacidify the vagina at the rate observed postcoitally following neutralization by the male ejaculate (W. H. Masters and V. E. Johnson, Human sexual response, p. 93, 1966). During bacterial vaginosis (BV), there is a loss of vaginal acidity, and the vaginal pH rises to >4.5. This correlates with a loss of lactobacilli and an overgrowth of diverse bacteria. Three BV-associated bacteria, Gardnerella vaginalis, Prevotella bivia, and Peptostreptococcus anaerobius, acidified their growth medium to an asymptotic pH (4.7 to 6.0) consistent with the characteristic elevated vaginal pH associated with BV. Together, these observations are consistent with vaginal flora, rather than epithelial cells, playing a primary role in creating the acidity of the vagina.

Boskey, E. R.; Telsch, K. M.; Whaley, K. J.; Moench, T. R.; Cone, R. A.

1999-01-01

230

Renal epithelial cells can release ATP by vesicular fusion  

PubMed Central

Renal epithelial cells have the ability to release nucleotides as paracrine factors. In the intercalated cells of the collecting duct, ATP is released by connexin30 (cx30), which is selectively expressed in this cell type. However, ATP is released by virtually all renal epithelia and the aim of the present study was to identify possible alternative nucleotide release pathways in a renal epithelial cell model. We used MDCK (type1) cells to screen for various potential ATP release pathways. In these cells, inhibition of the vesicular H+-ATPases (bafilomycin) reduced both the spontaneous and hypotonically (80%)-induced nucleotide release. Interference with vesicular fusion using N-ethylamide markedly reduced the spontaneous nucleotide release, as did interference with trafficking from the endoplasmic reticulum to the Golgi apparatus (brefeldin A1) and vesicular transport (nocodazole). These findings were substantiated using a siRNA directed against SNAP-23, which significantly reduced spontaneous ATP release. Inhibition of pannexin and connexins did not affect the spontaneous ATP release in this cell type, which consists of ~90% principal cells. TIRF-microscopy of either fluorescently-labeled ATP (MANT-ATP) or quinacrine-loaded vesicles, revealed that spontaneous release of single vesicles could be promoted by either hypoosmolality (50%) or ionomycin. This vesicular release decreased the overall cellular fluorescence by 5.8 and 7.6% respectively. In summary, this study supports the notion that spontaneous and induced ATP release can occur via exocytosis in renal epithelial cells.

Bjaelde, Randi G.; Arnadottir, Sigrid S.; Overgaard, Morten T.; Leipziger, Jens; Praetorius, Helle A.

2013-01-01

231

Immunohistochemical demonstration of airway epithelial cell markers of Guinea pig  

PubMed Central

The guinea pig (Cavea porcellus) is a mammalian non-rodent species in the Caviidae family. The sensitivity of the respiratory system and the susceptibility to infectious diseases allows the guinea pig to be a useful model for both infectious and non-infectious lung diseases such as asthma and tuberculosis. In this report, we demonstrated for the first time, the major cell types and composition in the guinea pig airway epithelium, using cell type-specific markers by immunohistochemical staining using the commercial available immunological reagents that cross-react with guinea pig. Our results revealed the availability of antibodies cross-reacting with airway epithelial cell types of basal, non-ciliated columnar, ciliated, Clara, goblet and alveolar type II cells, as well as those cells expressing Mucin 5AC, Mucin 2, Aquaporin 4 and Calcitonin Gene Related Peptide. The distribution of these various cell types were quantified in the guinea pig airway by immunohistochemical staining and were comparable with morphometric studies using an electron microscopy assay. Moreover, this study also demonstrated that goblet cells are the main secretory cell type in the guinea pig's airway, distinguishing this species from rats and mice. These results provide useful information for the understanding of airway epithelial cell biology and mechanisms of epithelial–immune integration in guinea pig models.

Li, Yong; Wang, Jing; He, Hai Yan; Ma, Ling Jie; Zeng, Jin; Deng, Guang Cun; Liu, Xiaoming; Engelhardt, John F.; Wang, Yujiong

2013-01-01

232

Human epithelial cell death caused by Actinobacillus actinomycetemcomitans infection.  

PubMed

The gingival sulcus is the shallow crevice around the tooth, and its epithelium is a gateway for initial bacterial infection in periodontal disease. Recent studies have shown that Actinobacillus actinomycetemcomitans invades an epithelial cell line, KB cells, in vitro. The aim of the present study was to clarify the changes in KB cells after A. actinomycetemcomitans infection. The cytotoxic effects of A. actinomycetemcomitans on KB cells were determined at 72, 96 and 120 h after infection by an MTT assay. Nuclear morphological changes were observed by staining with Hoechst 33258. Cytoplasmic histone-associated DNA fragmentation in the infected KB cells was determined by ELISA. A. actinomycetemcomitans was cytotoxic on KB cells, and condensation and degradation of the nuclei were observed. DNA fragmentation was increased after the infection. In addition, A. actinomycetemcomitans showed similar cytotoxic effects on human gingival epithelial cells. The present study demonstrated that A. actinomycetemcomitans induces apoptotic cell death of oral epithelial cells in an in-vitro culture system. This induced apoptosis might be involved in the initiation and progression of periodontitis. PMID:10933260

Kato, S; Nakashima, K; Inoue, M; Tomioka, J; Nonaka, K; Nishihara, T; Kowashi, Y

2000-08-01

233

Molecular characterization of TGF?-induced epithelial-mesenchymal transition in normal finite lifespan human mammary epithelial cells  

Microsoft Academic Search

Epithelial-mesenchymal transition (EMT) is a morphogenetic program essential for embryonic development and wound healing, but can adversely cause fibrosis and metastatic cancer progression when deregulated. Here, we established a model of efficient EMT induction in normal finite lifespan human mammary epithelial cells (HMEC) using transforming growth factor beta (TGF?). We demonstrate that EMT in HMEC occurs in three distinctive phases

Linsey E. Lindley; Karoline J. Briegel

2010-01-01

234

Expression of proliferating cell nuclear antigen (PCNA) in oral submucous fibrosis, oral epithelial hyperkeratosis and oral epithelial dysplasia in Taiwan  

Microsoft Academic Search

To test whether the oral epithelia of oral submucous fibrosis (OSF), epithelial hyperkeratosis (EH) and epithelial dysplasia (ED) may have increased proliferative activity under the long-term exposure to areca quid ingredients and whether there is an increased expression of proliferating cell nuclear antigen (PCNA) in oral premalignant lesions with disease progression, we used an immunohistochemical technique with the mouse monoclonal

C. P. Chiang; M. J. Lang; B. Y. Liu; J. T. Wang; J. S. Leu; L. J. Hahn; M. Y. P. Kuo

2000-01-01

235

Ionizing Radiation Predisposes Nonmalignant Human Mammary Epithelial Cells to Undergo Transforming Growth Factor B-Induced Epithelial to Mesenchymal Transition  

Microsoft Academic Search

Transforming growth factor B1 (TGFB) is a tumor suppressor during the initial stage of tumorigenesis, but it can switch to a tumor promoter during neoplastic progression. Ionizing radiation (IR), both a carcinogen and a therapeutic agent, induces TGFB activation in vivo. We now show that IR sensitizes human mammary epithelial cells (HMEC) to undergo TGFB-mediated epithelial to mesenchymal transition (EMT).

Kumari L. Andarawewa; Anna C. Erickson; William S. Chou; Sylvain V. Costes; Philippe Gascard; Joni D. Mott; Mina J. Bissell; Mary Helen Barcellos-Hoff

2007-01-01

236

The influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats  

Microsoft Academic Search

To investigate the influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats and explore the mechanisms of this effect. Wistar rats were randomly divided into the following three groups: control, diabetes and fasudil-treatment. All rats were sacrificed after three months of feeding with or without fasudil treatment. Pathological changes to the glomeruli and renal

Ganlin Wu; Yafang Tu; Ruhan Jia

2010-01-01

237

Prostatic intra-epithelial neoplasia: Expression and location of proliferating cell nuclear antigen in epithelial, endothelial and stromal nuclei  

Microsoft Academic Search

The expression and location of proliferating cell nuclear antigen (PCNA) immunostaining in epithelial, endothelial and stromal nuclei were assessed in prostatic intra-epithelial neoplasia (PIN). It was then compared with patterns in benign lesions and in invasive adenocarcinomas of the prostate. The PCNA-positive nuclei showed homogeneous or granular types of staining, or a mixture of both, and a gradation in the

R. Montironi; C. Magi Galluzzi; L. Diamanti; I. Giannulis; E. Pisani; M. Scarpelli

1993-01-01

238

NITROTYROSINE ATTENUATES RSV-INDUCED INFLAMMATION IN AIRWAY EPITHELIAL CELLS  

EPA Science Inventory

Nitrotyrosine attenuates RSV-induced inflammation in airway epithelial cells. Joleen Soukup, Zuowei Li, Susanne Becker and Yuh-Chin Huang. NHEERL, ORD, USEPA, RTP, North Carolina, CEMALB, University of North Carolina, Chapel Hill, North Carolina Nitrotyrosine (NO2Tyr) is a...

239

Klebsiella pneumoniae triggers a cytotoxic effect on airway epithelial cells  

Microsoft Academic Search

BACKGROUND: Klebsiella pneumoniae is a capsulated Gram negative bacterial pathogen and a frequent cause of nosocomial infections. Despite its clinical relevance, little is known about the features of the interaction between K. pneumoniae and lung epithelial cells on a cellular level, neither about the role of capsule polysaccharide, one of its best characterised virulence factors, in this interaction. RESULTS: The

Victoria Cano; David Moranta; Enrique Llobet-Brossa; José Antonio Bengoechea; Junkal Garmendia

2009-01-01

240

Human epithelial cell immortalization as a step in carcinogenesis  

Microsoft Academic Search

Human epithelial cells encounter two senescence barriers that enforce a limited proliferative potential. A first barrier is mediated by the retinoblastoma protein, and can be overcome by multiple types of errors, many of which are observed in human cancers. A second, extremely stringent telomere-dependent barrier, is a consequence of repression of telomerase activity. Although relieved by ectopic hTERT expression, the

Martha R. Stampfer; Paul Yaswen

2003-01-01

241

Adherence of Peptostreptococcus micros morphotypes to epithelial cells in vitro.  

PubMed

Peptostreptococcus micros, which is associated with oral and non-oral mixed anaerobic infections, occurs in three colony morphotypes, the smooth type, the rough type and the smooth variant of the rough type. These types differ in surface structures; the rough type expresses large fibrillar surface appendages, which are absent on the surface of both the smooth and the smooth variant of the rough type. To determine the role of these surface structures in adherence we characterized the adherence of the three morphotypes of P. micros to epithelial cells in vitro. Although all three types adhered well to epithelial cells, adhering numbers of the rough type were significantly lower than those of the smooth and the smooth variant of the rough type. Protease treatment increased the adherence of the rough type of the level of the two other types. The adherence of all three types was reduced more than 85% by treatment with 10 mM sodium periodate. Furthermore, the adherence was pH independent and could not be blocked by incubation with antisera to the bacteria. In addition, we determined the capacity to invade epithelial cells by P. micros. In an acridine orange assay such invasion could not be detected. Our results suggest that the adherence of P. micros to epithelial cells is mediated by periodate-sensitive extracellular polysaccharides and that the protruding fibril-like protein surface structures of the rough type have an obstructive effect on the adherence. PMID:10204480

Kremer, B H; Herscheid, A J; Papaioannou, W; Quirynen, M; van Steenbergen, T J

1999-02-01

242

Ultrastructural Study of Epithelial Cell Attachment to Implant Materials  

Microsoft Academic Search

Attachment of gingival tissues to the protruding post of an endosseous dental implant is of great importance for the prognosis of its clinical success. Little is known about the requirements an implant material must meet to secure a durable permucosal seal. The objective of this study, therefore, was to gain insight into the morphology of epithelial cell-implant interfaces for a

J. A. Jansen; J. R. De Wijn; J. M. L. Wolters-Lutgerhorst; P. J. Van Mullem

1985-01-01

243

Transcriptional profiling of mucociliary differentiation in human airway epithelial cells.  

PubMed

When cultured at an air-liquid interface (ALI) in the appropriate medium, primary human airway epithelial cells form a polarized, pseudostratified epithelium composed of ciliated and mucus-secreting cells. This culture system provides a useful tool for the in vitro study of airway epithelial biology and differentiation. We have performed microarray analysis on ALI cultures of human bronchial epithelial cells (HBECs) grown over a 28-d period to identify genes involved in mucociliary differentiation. We identified over 2,000 genes that displayed statistically significant 2-fold or greater changes in expression during the time course. Of the genes showing the largest increases, many are involved in processes associated with airway epithelial biology, such as cell adhesion, immunity, transport, and cilia formation; however, many novel genes were also identified. We compared our results with data from proteomic analyses of the ciliary axoneme and identified candidate genes that may have roles in cilia formation or function. Gene networks were generated using Ingenuity Pathways Analysis (Ingenuity Systems, Redwood City, CA) to identify signaling pathways involved in mucociliary cell differentiation or function. Networks containing genes involved in TGF-beta, WNT/beta-catenin, and epidermal growth factor receptor (EGFR) pathways were identified, suggesting potential roles for these families in airway epithelia. Microarray results were validated by real-time RT-PCR for a number of representative genes. This work has provided extensive information about gene expression changes during differentiation of airway epithelial cells, and will be a useful resource for researchers interested in respiratory function, pathology, and toxicology. PMID:17413031

Ross, Andrea J; Dailey, Lisa A; Brighton, Luisa E; Devlin, Robert B

2007-04-05

244

Human nasal and tracheo-bronchial respiratory epithelial cell culture.  

PubMed

Human airway epithelial (hAE) cell cultures are instrumental for studying basic and applied aspects of respiratory tract biology, disease, and therapy. When primary epithelial cells from the human nasal passages or tracheo-bronchial airways are grown on porous supports at an air-liquid interface (ALI) they undergo mucociliary differentiation, reproducing both the in vivo morphology and key physiologic processes. These cultures are useful for studying basic biology, disease pathogenesis, gene therapy and aerosol administration of drugs. This chapter gives detailed protocols for tissue procurement, cell isolation, production of complex media, and cell culture initiation and maintenance needed for hAE cell ALI cultures with non-proprietary reagents. PMID:23097104

Fulcher, M Leslie; Randell, Scott H

2013-01-01

245

In vitro culture and expansion of human limbal epithelial cells.  

PubMed

Limbal stem cells (LSCs) have an important role in the maintenance of the corneal surface epithelium, and autologous cultured limbal epithelial cell transplantations have contributed substantially to the treatment of the visually disabling condition known as LSC deficiency. In this protocol, we describe a method of establishing human limbal epithelial cell cultures by a feeder-free explant culture technique using a small limbal biopsy specimen and human amniotic membrane (hAM) as the culture substrate. This protocol is free of animal-derived products and involves the use of human recombinant growth factors. In addition, the recombinant cell dissociation enzyme TrypLE is used to replace trypsin and autologous serum replaces FBS. It takes approximately 2 weeks to establish a confluent monolayer from which approximately 3 x 10(6) cells can be harvested. This procedure can be adopted for both basic research purposes and clinical applications. PMID:20671730

Mariappan, Indumathi; Maddileti, Savitri; Savy, Soumya; Tiwari, Shubha; Gaddipati, Subhash; Fatima, Anees; Sangwan, Virender S; Balasubramanian, Dorairajan; Vemuganti, Geeta K

2010-07-29

246

The similarity between human embryonic stem cell-derived epithelial cells and ameloblast-lineage cells.  

PubMed

This study aimed to compare epithelial cells derived from human embryonic stem cells (hESCs) to human ameloblast-lineage cells (ALCs), as a way to determine their potential use as a cell source for ameloblast regeneration. Induced by various concentrations of bone morphogenetic protein 4 (BMP4), retinoic acid (RA) and lithium chloride (LiCl) for 7 days, hESCs adopted cobble-stone epithelial phenotype (hESC-derived epithelial cells (ES-ECs)) and expressed cytokeratin 14. Compared with ALCs and oral epithelial cells (OE), ES-ECs expressed amelogenesis-associated genes similar to ALCs. ES-ECs were compared with human fetal skin epithelium, human fetal oral buccal mucosal epithelial cells and human ALCs for their expression pattern of cytokeratins as well. ALCs had relatively high expression levels of cytokeratin 76, which was also found to be upregulated in ES-ECs. Based on the present study, with the similarity of gene expression with ALCs, ES-ECs are a promising potential cell source for regeneration, which are not available in erupted human teeth for regeneration of enamel. PMID:23538640

Zheng, Li-Wei; Linthicum, Logan; DenBesten, Pamela K; Zhang, Yan

2013-03-29

247

Radical-containing ultrafine particulate matter initiates epithelial-to-mesenchymal transitions in airway epithelial cells.  

PubMed

Environmentally persistent free radicals (EPFRs) in combustion-generated particulate matter (PM) are capable of inducing pulmonary pathologies and contributing to the development of environmental asthma. In vivo exposure of infant rats to EPFRs demonstrates their ability to induce airway hyperresponsiveness to methacholine, a hallmark of asthma. However, the mechanisms by which combustion-derived EPFRs elicit in vivo responses remain elusive. In this study, we used a chemically defined EPFR consisting of approximately 0.2 ?m amorphrous silica containing 3% cupric oxide with the organic pollutant 1,2-dichlorobenzene (DCB-230). DCB-230 possesses similar radical content to urban-collected EPFRs but offers several advantages, including lack of contaminants and chemical uniformity. DCB-230 was readily taken up by BEAS-2B and at high doses (200 ?g/cm(2)) caused substantial necrosis. At low doses (20 ?g/cm(2)), DCB-230 particles caused lysosomal membrane permeabilization, oxidative stress, and lipid peroxidation within 24 hours of exposure. During this period, BEAS-2B underwent epithelial-to-mesenchymal transition (EMT), including loss of epithelial cell morphology, decreased E-cadherin expression, and increased ?-smooth muscle actin (?-SMA) and collagen I production. Similar results were observed in neonatal air-liquid interface culture (i.e., disruption of epithelial integrity and EMT). Acute exposure of infant mice to DCB-230 resulted in EMT, as confirmed by lineage tracing studies and evidenced by coexpression of epithelial E-cadherin and mesenchymal ?-SMA proteins in airway cells and increased SNAI1 expression in the lungs. EMT in neonatal mouse lungs after EPFR exposure may provide an explanation for epidemiological evidence supporting PM exposure and increased risk of asthma. PMID:23087054

Thevenot, Paul T; Saravia, Jordy; Jin, Nili; Giaimo, Joseph D; Chustz, Regina E; Mahne, Sarah; Kelley, Matthew A; Hebert, Valeria Y; Dellinger, Barry; Dugas, Tammy R; Demayo, Francesco J; Cormier, Stephania A

2012-10-18

248

A novel closed cell culture device for fabrication of corneal epithelial cell sheets.  

PubMed

Automation technology for cell sheet-based tissue engineering would need to optimize the cell sheet fabrication process, stabilize cell sheet quality and reduce biological contamination risks. Biological contamination must be avoided in clinical settings. A closed culture system provides a solution for this. In the present study, we developed a closed culture device called a cell cartridge, to be used in a closed cell culture system for fabricating corneal epithelial cell sheets. Rabbit limbal epithelial cells were cultured on the surface of a porous membrane with 3T3 feeder cells, which are separate from the epithelial cells in the cell cartridges and in the cell-culture inserts as a control. To fabricate the stratified cell sheets, five different thicknesses of the membranes which were welded to the cell cartridge, were examined. Multilayered corneal epithelial cell sheets were fabricated in cell cartridges that were welded to a 25?µm-thick gas-permeable membrane, which was similar to the results with the cell-culture inserts. However, stratification of corneal epithelial cell sheets did not occur with cell cartridges that were welded to 100-300?µm-thick gas-permeable membranes. The fabricated cell sheets were evaluated by histological analyses to examine the expression of corneal epithelial-specific markers. Immunohistochemical analyses showed that a putative stem cell marker, p63, a corneal epithelial differentiation maker, CK3, and a barrier function marker, Claudin-1, were expressed in the appropriate position in the cell sheets. These results suggest that the cell cartridge is effective for fabricating corneal epithelial cell sheets. Copyright © 2012 John Wiley & Sons, Ltd. PMID:23239605

Nakajima, Ryota; Kobayashi, Toyoshige; Moriya, Noboru; Mizutani, Manabu; Kan, Kazutoshi; Nozaki, Takayuki; Saitoh, Kazuo; Yamato, Masayuki; Okano, Teruo; Takeda, Shizu

2012-12-14

249

Use of Percoll Density Gradients for Studying the Attachment of Bacteria to Oral Epithelial Cells  

Microsoft Academic Search

An assay for studying the attachment of bacteria to oral epithelial cells has been developed which utilizes Percoll density gradient centrifugation to separate bacteria and epithelial cells. 3H-thymidinelabeled bacteria were incubated with suspensions of buccal epithelial cells in microtitration plates for 2.5 hr at 35°C. The mixtures were then subjected to density gradient centrifugation in 50% Percoll. Epithelial cells with

W. C. Childs; R. J. Gibbons

1988-01-01

250

Vaginal Yeast Infections  

MedlinePLUS

... HIV/AIDS Sexually transmitted infections fact sheet Vaginal yeast infections fact sheet What is a vaginal yeast ... on vaginal yeast infections What is a vaginal yeast infection? A vaginal yeast infection is irritation of ...

251

Cadherin and integrin regulation of epithelial cell migration  

PubMed Central

These studies quantified the relative effects of E-cadherin expression and homophilic ligation on the integrin-mediated motility of epithelial cells. Micropatterned proteins were used to quantitatively titrate the ligation of E-cadherin and integrin receptors, in order to assess their coordinate influence on the migration velocities of MDA-MB-231 breast tumor epithelial cells. Fibronectin, E-cadherin, and mixtures of fibronectin and E-cadherin were covalently patterned on solid surfaces at defined compositions and mass coverages. The migration velocities of parental epithelial cells and of cells engineered to express E-cadherin under tetracycline control show that E-cadherin expression reduces cell motility by both adhesion-dependent and adhesion-independent mechanisms. Increasing E-cadherin expression levels also suppresses the dependence of cell velocity on the fibronectin coverage. On E-cadherin-containing substrata, the cell velocity decreases both with the E-cadherin expression level and with the immobilized E-cadherin surface density. These studies thus identified conditions under which E-cadherin preferentially suppresses cell migration by adhesion independent versus adhesion dependent mechanisms.

Silvestre, Jonathan; Kenis, Paul J.A.; Leckband, Deborah E.

2013-01-01

252

K+ channel inhibition accelerates intestinal epithelial cell wound healing.  

PubMed

Restitution is the process by which superficial interruptions in the gastrointestinal mucosa are repaired by the flattening and spreading of epithelial cells surrounding the damage. During this process, mucosal epithelial cells undergo extensive reshaping and cytoskeletal remodeling. K(+) channels, located primarily on the basolateral surface of gut epithelial cells, are central to both actin polymerization, via their control of membrane potential, and cell volume regulation. We questioned whether K(+) channels are involved in restitution using an in vitro model of intestinal epithelium, monolayers of the human colon carcinoma cell line T84. We report that pharmacologic K(+) channel inhibition accelerates wound healing in T84 cell monolayers. Both Ca(++)-dependent and constitutively active channels are involved, as indicated by the sensitivity to clotrimazole, charybdotoxin, and barium. The ability of clotrimazole to accelerate wound resealing was also observed in Caco-2 cell sheets. Pharmacologic stimulation of K(+) channel activity had no effect on the repair rate. Analysis of the resealing process by time lapse and confocal microscopy revealed that K(+) channel inhibitors abolished the initial wound retraction, briefly accelerated the repair rate, and altered the shape of the cell sheet abutting the injury during the early phase of resealing. We hypothesize that K(+) channel inactivation interrupts the coregulation of f-actin polymerization and volume control that is initiated by the healing process. PMID:15453839

Lotz, Margaret M; Wang, Helen; Song, Jaekyung Cecilia; Pories, Susan E; Matthews, Jeffrey B

253

Collective Epithelial and Mesenchymal Cell Migration During Gastrulation  

PubMed Central

Gastrulation, the process that puts the three major germlayers, the ectoderm, mesoderm and endoderm in their correct topological position in the developing embryo, is characterised by extensive highly organised collective cell migration of epithelial and mesenchymal cells. We discuss current knowledge and insights in the mechanisms controlling these cell behaviours during gastrulation in the chick embryo. We discuss several ideas that have been proposed to explain the observed large scale vortex movements of epithelial cells in the epiblast during formation of the primitive streak. We review current insights in the control and execution of the epithelial to mesenchymal transition (EMT) underlying the formation of the hypoblast and the ingression of the mesendoderm cells through the streak. We discuss the mechanisms by which the mesendoderm cells move, the nature and dynamics of the signals that guide these movements, as well as the interplay between signalling and movement that result in tissue patterning and morphogenesis. We argue that instructive cell-cell signaling and directed chemotactic movement responses to these signals are instrumental in the execution of all phases of gastrulation.

Chuai, Manli; Hughes, David; Weijer, Cornelis J

2012-01-01

254

Differentiation of cultured epithelial cells: Response to toxic agents  

SciTech Connect

Cell culture systems are instrumental in elucidating regulation of normal function and mechanisms of its perturbation by toxic substances. To this end, three applications of epithelial cells cultured with 3T3 feeder layer support are described. First, treatment of the premalignant human epidermal keratinocyte line SCC-12F2 with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate suppressed cell growth and differentiation. This agent produced a biphasic growth response greatly inhibiting cell growth at 1 to 10 nM, but much less above 100 nM. Expression of the differentiated functions involucrin and transglutaminase was found to be inhibited markedly at concentrations above 10 nM. Second, 3-methylcholanthrene toxicity was surveyed in a variety of rat epithelial cell types. The two most sensitive to growth inhibition were epidermal and mammary epithelial cells, while those from bladder, prostate, thyroid, and endometrium were insensitive to growth inhibition. Finally, expression of estrogen receptors in rat endometrial cells was shown to be stimulated by the cAmP-elevating agent forskolin. Maximal stimulation of 3- to 6-fold occurred in 6 hr, compatible with a requirement for protein synthesis. Pursuit of such results will aid in understanding differences in response among cell types and species, in elucidating mechanisms of action of known toxic substances and, ultimately, in predicting toxicity of less well understood agents.

Rice, R.H.; LaMontagne, A.D.; Petito, C.T.; Rong, Xianhui (Charles A. Dana Laboratory of Toxicology, Boston, MA (USA))

1989-03-01

255

Hexagonal Packing of Drosophila Wing Epithelial Cells by the Planar Cell Polarity Pathway  

Microsoft Academic Search

Summary The mechanisms that order cellular packing geometry are critical for the functioning of many tissues, but they are poorly understood. Here, we investigate this problem in the developing wing of Drosophila. The surface of the wing is decorated by hexagonally packed hairs that are uniformly oriented by the planar cellpolaritypathway.Theyareconstructedbyahexag- onal array of wing epithelial cells. Wing epithelial cells

Anne-Kathrin Classen; Kurt I. Anderson; Eric Marois; Suzanne Eaton

2005-01-01

256

Bone marrow mesenchymal stem cells can differentiate into type II alveolar epithelial cells in vitro.  

PubMed

In this study, we demonstrate that BMSCs (bone marrow mesenchymal stem cells) can be successfully differentiated into type II alveolar epithelial cells in vitro under mimic pulmonary microenvironment. BMSCs were co-cultured with MRC-5 cells in modified SAGM (small airway growth medium). The BMSC-derived type II alveolar epithelial cells morphologically resemble human lung epithelial cells. They began to appear after 10 days in co-culture and became morphologically dominant after day 15. Correspondingly, SPC (surfactant protein C), a specific functional marker of human type II alveolar epithelial cells, was detected in differentiated cells by RT-PCR (reverse transcription-PCR) analysis after day 15. Immunostaining analysis revealed the present of scattered SPC-positive cells with a differentiation efficiency of 2.43-4.21%. Our study further showed that the SPC gene expression level in differentiated cells was related to the ratio of BMSCs to MRC-5 cells and the components of modified SAGM. PMID:21542803

Ma, Nan; Gai, Hui; Mei, Ju; Ding, Fang-Bao; Bao, Chun-Rong; Nguyen, David M; Zhong, Hong

2011-12-01

257

Isolation and characterization of mouse and human esophageal epithelial cells in 3D organotypic culture  

Microsoft Academic Search

This protocol describes the isolation and characterization of mouse and human esophageal epithelial cells and the application of 3D organotypic culture (OTC), a form of tissue engineering. This model system permits the interrogation of mechanisms underlying epithelial-stromal interactions. We provide guidelines for isolating and cultivating several sources of epithelial cells and fibroblasts, as well as genetic manipulation of these cell

Jiri Kalabis; Gabrielle S Wong; Maria E Vega; Mitsuteru Natsuizaka; Erle S Robertson; Meenhard Herlyn; Hiroshi Nakagawa; Anil K Rustgi

2012-01-01

258

Arkadia regulates TGF-? signaling during renal tubular epithelial to mesenchymal cell transition  

Microsoft Academic Search

Transforming growth factor-? (TGF-?) signaling has been linked with tubular epithelial to mesenchymal cell transition. In this study, we examined the role of Arkadia, an E3 ubiquitin ligase that is critically required for TGF-? signaling during epithelial to mesenchymal cell transition. We found that when normal human renal tubular epithelial cells in culture were stimulated with TGF-?1, which increased their

F-Y Liu; X-Z Li; Y-M Peng; H Liu; Y-H Liu

2008-01-01

259

The fate of Treponema denticola within human gingival epithelial cells.  

PubMed

Treponema denticola is one of the major pathogens associated with chronic periodontitis. Bacterial invasion into gingival tissues is a critical process in the pathogenesis of periodontal disease. We recently reported that T. denticola can invade gingival epithelial cells. The aim of this study is to determine the fate of internalized T. denticola in gingival epithelial cells. Immortalized human gingival epithelial HOK-16B cells were infected with 5- (and 6-) carboxy-fluorescein diacetate succinimidyl ester (CFSE)-labeled live or heat-killed T. denticola for 24 h, and the presence of bacteria inside the cells was confirmed by confocal microscopy. Live T. denticola, but not heat-killed bacteria, invaded HOK-16B cells. Confocal microscopy also revealed that internalized T. denticola rarely colocalized with either endosomes or lysosomes. Transmission electron microscopy of infected cells showed that intracellular T. denticola was localized inside endosome-like structures. Although a culture-based antibiotics protection assay could not detect viable intracellular T. denticola 12 h after infection, a substantial number of bacteria were observed by confocal microscopy and weak expression of bacterial 16S ribosomal RNA was detected 48 h after infection. In addition, flow cytometric analysis of HOK-16B cells infected with CFSE-labeled T. denticola showed no loss of fluorescence over 48 h. Collectively, T. denticola invades gingival epithelial cells and remains within the host cells for many hours by resisting endolysosomal degradation. These findings may provide new insight into the role of T. denticola in the pathogenesis of periodontitis. PMID:23134612

Shin, J; Choi, Y

2012-07-21

260

Modulation of epithelial tissue and cell migration by microgrooves.  

PubMed

We used a polystyrene substratum to study the response of migrating epithelium to 1- or 5-microm depth microgrooves with groove/ridge widths of 1, 2, 5, or 10 microm. The migration of a tissue sheet was enhanced along the microgrooves, while migration across the microgrooves was inhibited. Changing the depth of the microgrooves had a greater effect on migration than alteration of the groove/ridge width. The migration of epithelial cells from a confluent monolayer culture followed a similar pattern to that of intact epithelial tissue. Cellular extensions generally followed the microgroove direction by tracking along the top of the ridges or following the ridge walls, as revealed by scanning electron microscopy. Actin filaments within the basal cell layer of the tissue were aligned with the microgrooves, unlike filaments in the superficial layers that did not appear to be affected by the presence of underlying microgrooves. The basal cell layer of the tissue conformed to the contours of the microgroove following migration. However, the ultrastructure of the tissue above the ridges resembled that of tissue on a flat surface. We concluded that surface microgrooves have the potential to direct the migration of immediately adjacent epithelial tissue, the effect of which is to guide epithelial tissue on the surface of implanted biomaterials. PMID:11340589

Dalton, B A; Walboomers, X F; Dziegielewski, M; Evans, M D; Taylor, S; Jansen, J A; Steele, J G

2001-08-01

261

Lectin-resistant mutants of polarized epithelial cells.  

PubMed

Two lectin-resistant mutants derived from Madin Darby canine kidney cells, with constitutive alterations in the asparagine-linked carbohydrate moieties, retained the characteristic structural and functional epithelial polarity of the parental cells. A ricin-resistant cell line was unable to incorporate galactose-sialic acid into glycoproteins and, from the pattern of cross-resistance to other lectins, appears to be different from previously described lines resistant to this lectin: the mutation in a concanavalin A-resistant line results, probably, in the production of defective carbohydrate cores of glycoproteins. In spite of glycosylation defects which result in an increased electrophoretic mobility of many cellular glycoproteins, both mutants retained the typical asymmetric structure of the plasma membrane (microvilli on the apical surface, junctional elements on the basolateral surface), functional tight junctions, and unidirectional active transport of electrolytes and water. These results suggest that glycoproteins with terminal galactose-sialic acid moieties are not critically involved in the development and maintenance of polarity in epithelial cells. The mutant cells, particularly the ricin-resistant line, exhibited, however, morphological and electrophysiological changes which suggest a quantitative effect of the mutations on intracellular traffic of membranes and tight junction formation. The cell lines described in this paper, the first lectin-resistant mutants of epithelial lineage, should prove useful tools for studying the peculiarities of glycosylating pathways in polarized cells. PMID:7177111

Meiss, H K; Green, R F; Rodriguez-Boulan, E J

1982-10-01

262

Collective and single cell behavior in epithelial contact inhibition  

PubMed Central

Control of cell proliferation is a fundamental aspect of tissue physiology central to morphogenesis, wound healing, and cancer. Although many of the molecular genetic factors are now known, the system level regulation of growth is still poorly understood. A simple form of inhibition of cell proliferation is encountered in vitro in normally differentiating epithelial cell cultures and is known as “contact inhibition.” The study presented here provides a quantitative characterization of contact inhibition dynamics on tissue-wide and single cell levels. Using long-term tracking of cultured Madin-Darby canine kidney cells we demonstrate that inhibition of cell division in a confluent monolayer follows inhibition of cell motility and sets in when mechanical constraint on local expansion causes divisions to reduce cell area. We quantify cell motility and cell cycle statistics in the low density confluent regime and their change across the transition to epithelial morphology which occurs with increasing cell density. We then study the dynamics of cell area distribution arising through reductive division, determine the average mitotic rate as a function of cell size, and demonstrate that complete arrest of mitosis occurs when cell area falls below a critical value. We also present a simple computational model of growth mechanics which captures all aspects of the observed behavior. Our measurements and analysis show that contact inhibition is a consequence of mechanical interaction and constraint rather than interfacial contact alone, and define quantitative phenotypes that can guide future studies of molecular mechanisms underlying contact inhibition.

Puliafito, Alberto; Hufnagel, Lars; Neveu, Pierre; Streichan, Sebastian; Sigal, Alex; Fygenson, D. Kuchnir; Shraiman, Boris I.

2012-01-01

263

Intercellular communication in cultured rabbit gastric epithelial cells.  

PubMed

The effects of drugs related to cyclic AMP and a tumor promoter, phorbol ester, on intercellular communications via gap junctions were investigated by the Lucifer Yellow-transfer method in cultured rabbit gastric epithelial cells. Cells were in contact with each drug for 4 hr before the microinjection of the dye into a cell. Dye transfer capacity was significantly increased by dibutyryl cyclic AMP (10(-3) M), theophylline (10(-3) M), 3-isobutyl-1-methylxanthine (10(-4) M), forskolin (10(-6) M) and irsogladine (10(-4) M); and it was inhibited by 12-O-tetradecanoyl-phorbol-13-acetate (100 ng/ml). These results suggest that the intercellular communication between cultured rabbit gastric epithelial cells is upregulated by cyclic AMP. PMID:1667533

Ueda, F; Kyoi, T; Mimura, K; Kimura, K; Yamamoto, M

1991-11-01

264

Inactivation of Rb in stromal fibroblasts promotes epithelial cell invasion  

PubMed Central

Stromal-derived growth factors are required for normal epithelial growth but are also implicated in tumour progression. We have observed inactivation of the retinoblastoma protein (Rb), through phosphorylation, in cancer-associated fibroblasts in oro-pharyngeal cancer specimens. Rb is well known for its cell-autonomous effects on cancer initiation and progression; however, cell non-autonomous functions of Rb are not well described. We have identified a cell non-autonomous role of Rb, using three-dimensional cultures, where depletion of Rb in stromal fibroblasts enhances invasive potential of transformed epithelia. In part, this is mediated by upregulation of keratinocyte growth factor (KGF), which is produced by the depleted fibroblasts. KGF drives invasion of epithelial cells through induction of MMP1 expression in an AKT- and Ets2-dependent manner. Our data identify that stromal fibroblasts can alter the invasive behaviour of the epithelium, and we show that altered expression of KGF can mediate these functions.

Pickard, Adam; Cichon, Ann-Christin; Barry, Anna; Kieran, Declan; Patel, Daksha; Hamilton, Peter; Salto-Tellez, Manuel; James, Jacqueline; McCance, Dennis J

2012-01-01

265

Inactivation of Rb in stromal fibroblasts promotes epithelial cell invasion.  

PubMed

Stromal-derived growth factors are required for normal epithelial growth but are also implicated in tumour progression. We have observed inactivation of the retinoblastoma protein (Rb), through phosphorylation, in cancer-associated fibroblasts in oro-pharyngeal cancer specimens. Rb is well known for its cell-autonomous effects on cancer initiation and progression; however, cell non-autonomous functions of Rb are not well described. We have identified a cell non-autonomous role of Rb, using three-dimensional cultures, where depletion of Rb in stromal fibroblasts enhances invasive potential of transformed epithelia. In part, this is mediated by upregulation of keratinocyte growth factor (KGF), which is produced by the depleted fibroblasts. KGF drives invasion of epithelial cells through induction of MMP1 expression in an AKT- and Ets2-dependent manner. Our data identify that stromal fibroblasts can alter the invasive behaviour of the epithelium, and we show that altered expression of KGF can mediate these functions. PMID:22643222

Pickard, Adam; Cichon, Ann-Christin; Barry, Anna; Kieran, Declan; Patel, Daksha; Hamilton, Peter; Salto-Tellez, Manuel; James, Jacqueline; McCance, Dennis J

2012-05-29

266

Normal and tumour cervical cells respond differently to vaginal lactobacilli, independent of pH and lactate.  

PubMed

Cervical cancer is a human papilloma virus (HPV)-related cancer, but most HPV infections are transient or intermittent and resolve spontaneously. Thus, other factors, such as cervical microflora, which are dominated by lactobacilli, must be involved in invasive cervical carcinoma development after HPV infection. Previous studies have demonstrated that lactobacilli have antitumour effects, and it is possible that vaginal lactobacilli prevent cervical cancer. Here we examined the proliferative and apoptotic responses of normal and tumour cervical cells to common vaginal lactobacilli components by investigating human normal fibroblast-like cervical (normal cervical) and HeLa (cervical tumour) cell responses to Lactobacillus gasseri and Lactobacillus crispatus. The effects of different lactobacilli components, such as culture supernatants, cytoplasmic extracts, cell-wall extracts and live cells, were determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, trypan blue staining, lactate dehydrogenase assay and colorimetric caspase-3 activity assay. Changes in caspase-3 and human chorionic gonadotropin ? (hCG?) expression were analysed by quantitative RT-PCR. Tumour cell growth inhibition by culture supernatants was higher than that by pH- and lactate-adjusted controls. However, the effects of the supernatants on normal cells were similar to those of lactate-adjusted controls. Apoptosis was inhibited by supernatants, which was consistent with higher hCG? expression since hCG inhibits apoptosis. Our study demonstrated that common vaginal lactobacilli exert cytotoxic effects on cervical tumour cells, but not on normal cells, and that this cytotoxicity is independent of pH and lactate. Our results encourage further studies on the interaction between lactobacilli and cervical cells, and administration of common vaginal lactobacilli as probiotics. PMID:23618799

Motevaseli, Elahe; Shirzad, Mahdieh; Akrami, Seyed Mohammad; Mousavi, Azam-Sadat; Mirsalehian, Akbar; Modarressi, Mohammad Hossein

2013-04-25

267

Growth Factor Regulation of Cell Cycle Progression in Mammary Epithelial Cells  

Microsoft Academic Search

Growth factors are among the critical positive and negative regulators of cell proliferation for normal mammary\\/breast epithelial cells and for breast cancer cells. The mechanisms by which specific growth factors regulate the cell cycle in mammary\\/breast epithelial cells is beginning to be understood for several growth factor families, including the epidermal growth factor, insulin-like growth factor, and transforming growth factor-beta

Malinda A. Stull; Anne M. Rowzee; Aimee V. Loladze; Teresa L. Wood

2004-01-01

268

Production of Skeletal Muscle Elements by Cell Lines Derived from Neoplastic Rat Mammary Epithelial Stem Cells  

Microsoft Academic Search

Single-cell-cloned cell lines intermediate in morphology be tween the cuboidal epithelial and fully elongated myoepithelial- like cells have been isolated from the single-cell-cloned epithelial stem cell lines Rama 25 and Rama 37 originally obtained from dimethylbenz(a)anthracene-induced mammary tumors from Sprague-Dawley and Wistar-Furth rats, respectively. These are designated Rama 25-11, Rama 25-I2, Rama 25-I4 (Sprague- Dawley) and Rama 50-55, Rama 59,

Philip S. Rudland; Damien J. Dunnington; Barry Gusterson; Paul Monaghan; Christine M. Hughes

269

MicroRNA 16 Modulates Epithelial Sodium Channel in Human Alveolar Epithelial Cells  

PubMed Central

Acute lung injury (ALI) is a devastating disease characterized by pulmonary edema. Removal of edema from the air spaces is a critical function of the epithelial sodium channel (ENaC) in ALI. The molecular mechanisms behind resolution of pulmonary edema are incompletely understood. MicroRNA’s (miRNA) are crucial gene regulators and are dysregulated in various diseases including ALI. Recent studies suggest that microRNA-16 (miR-16) targets serotonin transporter (SERT) involved in the serotonin (5-HT) transmitter system. Alterations in serotonin levels have been reported in various pulmonary diseases. However, the role of miR-16 on its target SERT, and ENaC, a key ion channel involved in the resolution of pulmonary edema, have not been studied. In the present study, the expression patterns of miR-16, SERT, ENaC and serotonin were investigated in mice exposed to room air and hyperoxia. The effects of miR-16 overexpression on ENaC, SERT, TGF-? and Nedd4 in human alveolar epithelial cells were analyzed. miR-16 and ENaC were downregulated in mice exposed to hyperoxia. miR-16 downregulation in mouse lung was correlated with an increase in SERT expression and pulmonary edema. Overexpression of miR-16 in human alveolar epithelial cells (A549) suppressed SERT and increased ENaC? levels when compared to control-vector transfected cells. In addition, miR-16 over expression suppressed TGF? release, a critical inhibitor of ENaC. Interestingly Nedd4, a negative regulator of ENaC remained unaltered in miR-16 over expressed A549 cells when compared to controls. Taken together, our data suggests that miR-16 upregulates ENaC, a major sodium channel involved in resolution of pulmonary edema in ALI.

Parthasarathy, Prasanna Tamarapu; Galam, Lakshmi; Huynh, Bao; Yunus, Asfiya; Abuelenen, Toaa; Castillo, Annie; Ramanathan, Gurukumar Kollongod; Ruan, Cox; Kolliputi, Narasaiah

2012-01-01

270

Concise review: limbal epithelial stem cell therapy: controversies and challenges.  

PubMed

Limbal epithelial stem cells (LESCs) are a population of stem cells responsible for maintenance and repair of the corneal surface. Injury and disease can result in a deficiency of these stem cells, the vision affecting condition called limbal stem cell deficiency (LSCD) in which the cornea becomes opaque, vascularized, and inflamed. Cultured LESC therapy was first described in 1997;29:19231932-19231932.and LESCs cultured from either patients or donors have been used to successfully treat LSCD. In this review, some of the challenges and controversies associated with cultured LESC therapy will be discussed including alternative stem cell sources. PMID:21997829

O'Callaghan, Anna R; Daniels, Julie T

2011-12-01

271

Vaccination Against Tumor Cells Expressing Breast Cancer Epithelial Tumor Antigen  

Microsoft Academic Search

Ninety-one percent of breast tumors aberrantly express an epithelial tumor antigen (ETA) identified by monoclonal antibody H23. Vaccinia virus recombinants expressing tumor antigens have considerable promise in the active immunotherapy of cancer, and we have evaluated the potential of vaccinia recombinants expressing the secreted (S) and cell-associated (transmembrane, T) forms of H23 ETA to elicit immunity to tumor cells expressing

Mara Hareuveni; Claudie Gautier; Marie-Paule Kieny; Daniel Wreschner; Pierre Chambon; Richard Lathe

1990-01-01

272

Biological principals and clinical potentials of limbal epithelial stem cells  

Microsoft Academic Search

In this review, we describe a population of adult stem cells that are currently being successfully used in the clinic to treat\\u000a blinding ocular surface disease, namely limbal epithelial stem cells (LESC). The function and characteristics of LESC and\\u000a the challenges faced in making use of their therapeutic potential will be examined. The cornea on the front surface of the

Maria Notara; Julie T. Daniels

2008-01-01

273

Cytotoxic Action of Serratia marcescens Hemolysin on Human Epithelial Cells  

Microsoft Academic Search

Incubation of human epithelial cells with nanomolar concentrations of chromatographically purified Serratia marcescens hemolysin (ShlA) caused irreversible vacuolation and subsequent lysis of the cells. Vacuolation differed from vacuole formation by Helicobacter pylori VacA. Sublytic doses of ShlA led to a reversible depletion of intracellular ATP. Restoration to the initial ATP level was presumably due to the repair of the toxin

RALF HERTLE; MARTINA HILGER; SANDRA WEINGARDT-KOCHER

274

Polarized Distribution of Chloride Channels in Epithelial Cells  

Microsoft Academic Search

Cl-currents in cultured epithelial cells (MDCK) are inhibited by 4-acetamido-4’-isothiocyano-2,2’-disulfonic acid stilbene; 4,4’-diisothiocyanatostilbene-2,2’-disulfonic aicd, and diphenylamine-2-carboxylate are little affected by trypsin-EDTA, and exhibit a 260 % increase one day after plating at confluence. Although this increase in Cl-conductance coexists with an outburst of mitosis, it does not seem to be associated to the cell cycle, because (1) it is not

A. Ponce; R. G. Contreras; M. Cereijido

1991-01-01

275

Growth and characterization of human skin epithelial cell cultures  

Microsoft Academic Search

Summary  In 129 of 140 attempts, human skin cells were successfully cultured on the dermal collagen bed of sterile, dead pigskin. Diploid\\u000a epithelial cells grew selectively on the collagen bed; fibroblasts grew on the glass surfaces of the culture dishes. The cultures\\u000a could be subdivided physically up to six times at a 1:2 split ratio, but at least 24 to 48

Aaron E. Freeman; Howard J. Igel; Brenda J. Herrman; Karen L. Kleinfeld

1976-01-01

276

Fibronectin Synthesis by Epithelial Crypt Cells of Rat Small Intestine  

Microsoft Academic Search

Synthesis of fibronectin in an epithelial cell line (IEC-6) established from rat small intestine was demonstrated by using immunofluorescence, radioimmunoassay, and collagen-binding. Internally labeled radioactive fibronectin isolated from the IEC-6 cells gave a single main band in sodium dodecyl sulfate\\/polyacrylamide gel electrophoresis under reducing conditions. Fibronectin isolated from rat plasma gave two closely spaced bands. The slower one had the

Andrea Quaroni; Kurt J. Isselbacher; Erkki Ruoslahti

1978-01-01

277

Functional diversity of human vaginal APC subsets in directing T cell responses  

PubMed Central

Human vaginal mucosa is the major entry site of sexually transmitted pathogens and thus has long been attractive as a site for mounting mucosal immunity. It is also known as a tolerogenic microenvironment. Here, we demonstrate that immune responses in the vagina are orchestrated by the functional diversity of four major antigen-presenting cell (APC) subsets. Langerhans cells (LCs) and CD14? lamina propria (LP)-DCs polarize CD4+ and CD8+ T cells toward Th2, whereas CD14+ LP-DCs and macrophages polarize CD4+ T cells toward Th1. Both LCs and CD14? LP-DCs are potent inducers of Th22. Due to their functional specialties and the different expression levels of pattern-recognition receptors on the APC subsets, microbial products do not bias them to elicit common types of immune responses (Th1 or Th2). To evoke desired types of adaptive immune responses in the human vagina, antigens may need to be targeted to proper APC subsets with right adjuvants.

Duluc, Dorothee; Gannevat, Julien; Anguiano, Esperanza; Zurawski, Sandra; Carley, Michael; Boreham, Muriel; Stecher, Jack; Dullaers, Melissa; Banchereau, Jacques; Oh, SangKon

2012-01-01

278

Review: corneal epithelial stem cells, their niche and wound healing.  

PubMed

Stem cells emerged as a concept during the second half of 19(th) century, first as a theoretical entity, but then became one of the most promising research fields in cell biology. This work describes the most important characteristics of adult stem cells, including the experimental criteria used to identify them, and discusses current knowledge that led to the proposal that stem cells existed in different parts of the eye, such as the retina, lens, conjunctiva, corneal stroma, Descemet's membrane, and the subject of this review: the corneal epithelium. Evidence includes results that support the presence of corneal epithelial stem cells at the limbus, as well as the major obstacles to isolating them as pure cell populations. Part of this review describes the variation in the basement membrane composition between the limbus and the central cornea, to show the importance of the corneal stem cell niche, its structure, and the participation of extracellular matrix (ECM) components in regulating corneal stem cell compartment. Results obtained by various laboratories suggest that the extracellular matrix plays a central role in regulating stem cell commitment, corneal differentiation, and participation in corneal wound healing, in addition to other environmental signals such as cytokines and growth factors. The niche could define cell division patterns in corneal stem cell populations, establishing whether stem cells divide asymmetrically or symmetrically. Characterization and understanding of the factors that regulate corneal epithelial stem cells should open up new paths for developing new therapies and strategies for accelerating and improving corneal wound healing. PMID:23901244

Castro-Muñozledo, Federico

2013-07-24

279

Epithelial stem cells, wound healing and cancer  

Microsoft Academic Search

It is well established that tissue repair depends on stem cells and that chronic wounds predispose to tumour formation. However, the association between stem cells, wound healing and cancer is poorly understood. Lineage tracing has now shown how stem cells are mobilized to repair skin wounds and how they contribute to skin tumour development. The signalling pathways, including WNT and

Esther N. Arwert; Esther Hoste; Fiona M. Watt

2012-01-01

280

Hsc70 negatively regulates epithelial sodium channel trafficking at multiple sites in epithelial cells.  

PubMed

The epithelial sodium channel (ENaC) plays an important role in homeostasis of blood pressure and of the airway surface liquid, and excess function of ENaC results in refractory hypertension (in Liddle's syndrome) and impaired mucociliary clearance (in cystic fibrosis). The regulation of ENaC by molecular chaperones, such as the 70-kDa heat shock protein Hsc70, is not completely understood. Our previously published data suggest that Hsc70 negatively affects ENaC activity and surface expression in Xenopus oocytes; here we investigate the mechanism by which Hsc70 acts on ENaC in epithelial cells. In Madin-Darby canine kidney cells stably expressing epitope-tagged ???-ENaC and with tetracycline-inducible overexpression of Hsc70, treatment with 5 ?g/ml doxycycline increased total Hsc70 expression 20%. This increase in Hsc70 expression led to a decrease in ENaC activity and surface expression that corresponded to an increased rate of functional ENaC retrieval from the cell surface. In addition, Hsc70 overexpression decreased the association of newly synthesized ENaC subunits. These data support the hypothesis that Hsc70 inhibits ENaC functional expression at the apical surface of epithelia by regulating ENaC biogenesis and ENaC trafficking at the cell surface. PMID:23885065

Chanoux, Rebecca A; Shubin, Calla B; Robay, Amal; Suaud, Laurence; Rubenstein, Ronald C

2013-07-24

281

Neuroendocrinelike (small granule) epithelial cells of the lung.  

PubMed Central

The presence of neuroendocrinelike epithelial cells in the lung of numerous species has been demonstrated by light and electron microscopy. Histochemical methods used to identify these cells have included staining with silver, amine-type fluorescence (APUD cell), periodic acid Schiff (PAS)-lead hematoxylin, and immunohistochemical localization of neuron-specific enolase. Cytoplasmic dense core vesicles (70-200 nm in diameter) have served as the major ultrastructural characteristic. Lung neuroendocrinelike cells have been shown to occur in fetal and adult mammals as solitary-type cells or as distinct organoids known as neuroepithelial bodies ( NEBs ). Although the frequency of both populations is considered low, solitary-type cells with dense-core granules can be found in as high as 5% of epithelial cells in the cricoid region of the guinea-pig larynx. The solitary cells can be found throughout the airways of mammals, whereas the NEBs are confined to the intrapulmonary airways. Unmyelinated fibers have been traced from the lamina propria and into the NEB, where they ramified between the component cells of the NEB. The function of lung neuroendocrinelike cells is not known, but morphological and cytochemical studies suggest that the NEBs are intrapulmonary chemoreceptors that can respond to changes in airway gas composition. Hypoxia or hypercapnia has been shown to decrease the amine cytofluorescence in these organoids and apparently to increase the exocytosis of dense core vesicles from the basal region of the cell. Immunohistochemical studies have suggested that some lung epithelial cells may contain a known neuropeptide(s), but further investigation is needed to confirm the presence of such compounds in lung neuroendocrinelike cells and their physiochemical properties. Apparent hyperplasia of lung neuroendocrinelike cells can occur readily in hamsters treated with diethylnitrosamine. It has been postulated that human lung tumors with endocrinelike properties, namely, bronchial carcinoids and lung small cell carcinomas, may originate from lung neuroendocrinelike cells. However, a more plausible explanation, based on cytokinetic studies of epithelial neuroendocrinelike cells in the lung and other organs, is that these cells originate from a nonneuroendocrine population. Interaction of such a progenitor cell population with selected carcinogens may lead to stimulation of the rate of normal differentiation or, alternately, to selection of an abnormal route of differentiation that possesses a neuroendocrine phenotype. Images FIGURE 1. FIGURE 2. FIGURE 3. FIGURE 4. FIGURE 5. FIGURE 6. FIGURE 7. FIGURE 8. FIGURE 9. FIGURE 10. FIGURE 11. FIGURE 12. FIGURE 13. FIGURE 14. FIGURE 15.

DiAugustine, R P; Sonstegard, K S

1984-01-01

282

Bacterial Exposure Induces and Activates Matrilysin in Mucosal Epithelial Cells  

PubMed Central

Matrilysin, a matrix metalloproteinase, is expressed and secreted lumenally by intact mucosal and glandular epithelia throughout the body, suggesting that its regulation and function are shared among tissues. Because matrilysin is produced in Paneth cells of the murine small intestine, where it participates in innate host defense by activation of prodefensins, we speculated that its expression would be influenced by bacterial exposure. Indeed, acute infection (10–90 min) of human colon, bladder, and lung carcinoma cells, primary human tracheal epithelial cells, and human tracheal explants with type 1–piliated Escherichia coli mediated a marked (25–50-fold) and sustained (>24 h) induction of matrilysin production. In addition, bacterial infection resulted in activation of the zymogen form of the enzyme, which was selectively released at the apical surface. Induction of matrilysin was mediated by a soluble, non-LPS bacterial factor and correlated with the release of defensin-like bacteriocidal activity. Bacteria did not induce matrilysin in other cell types, and expression of other metalloproteinases by epithelial cells was not affected by bacteria. Matrilysin was not detected in germ-free mice, but the enzyme was induced after colonization with Bacteroides thetaiotaomicron. These findings indicate that bacterial exposure is a potent and physiologically relevant signal regulating matrilysin expression in epithelial cells.

Lopez-Boado, Yolanda S.; Wilson, Carole L.; Hooper, Lora V.; Gordon, Jeffrey I.; Hultgren, Scott J.; Parks, William C.

2000-01-01

283

Nesfatin-1 inhibits ovarian epithelial carcinoma cell proliferation in vitro.  

PubMed

Nesfatin-1, an 82-amino-acid peptide derived from a 396-amino-acid precursor protein nucleobindin 2 (NUCB2), was originally identified in hypothalamic nuclei involved in the regulation of food intake. It was recently reported that nesfatin-1 is a novel depot specific adipokine preferentially produced by subcutaneous tissue, with obesity- and food deprivation-regulated expression. Although a relation between ovarian cancer mortality and obesity has been previously established, a role of nesfatin-1 in ovarian epithelial carcinoma remains unknown. The aim of the present study is to examine the effect of nesfatin-1 on ovary carcinoma cells proliferation. We found that nesfatin-1 inhibits the proliferation and growth of HO-8910 cells by G1 phase arrest, this inhibition could be abolished by nesfatin-1 neutralizing antibody. Nesfatin-1 enhances HO-8910 cell apoptosis, activation of mammalian target of rapamycin (mTOR) and RhoA/ROCK signaling pathway block the effects of nesfatin-1-induced apoptosis, therefore reverses the inhibition of HO-8910 cell proliferation by nesfatin-1. In conclusion, the present study demonstrated that nesfatin-1 can inhibit the proliferation in human ovarian epithelial carcinoma cell line HO-8910 cells through inducing apoptosis via mTOR and RhoA/ROCK signaling pathway. This study provides a novel regulatory signaling pathway of nesfatin-1-regulated ovarian epithelial carcinoma growth and may contribute to ovarian cancer prevention and therapy, especially in obese patients. PMID:23764403

Xu, Yang; Pang, Xiaoyan; Dong, Mei; Wen, Fang; Zhang, Yi

2013-06-11

284

A novel method for preserving cultured limbal epithelial cells  

PubMed Central

Aim To investigate organ culture preservation of cultured limbal epithelial cells in order to enhance the availability of tissue?engineered epithelia that are used to treat patients with limbal stem cell deficiency. Methods Limbal epithelial cells were cultured for 3?weeks on intact amniotic membrane fastened to a polyester membrane carrier. The cultured epithelia were stored for 1?week at 23°C in organ culture medium. The preserved epithelia were then examined using a colorimetric cell viability assay, light microscopy and immunohistochemistry. Results The viability of the preserved epithelia was 84% (20%), and no statistically significant difference was found compared with non?preserved epithelia. In general, the cell borders were maintained, the nuclei showed no sign of degeneration, and the original layered structure was preserved. Mild intercellular oedema was occasionally observed. Expression of p63, K19 and vimentin was maintained. Conclusions Cultured limbal epithelial cells can be preserved in organ culture medium for 1?week at room temperature, while maintaining the original layered structure and undifferentiated phenotype.

Utheim, Tor Paaske; Raeder, Sten; Utheim, ?ygunn Aass; Cai, Yiqing; Roald, Borghild; Drolsum, Liv; Lyberg, Torstein; Nicolaissen, Bj?rn

2007-01-01

285

Established thymic epithelial progenitor/stem cell-like cell lines differentiate into mature thymic epithelial cells and support T cell development.  

PubMed

Common thymic epithelial progenitor/stem cells (TEPCs) differentiate into cortical and medullary thymic epithelial cells (TECs), which are required for the development and selection of thymocytes. Mature TEC lines have been widely established. However, the establishment of TEPC lines is rarely reported. Here we describe the establishment of thymic epithelial stomal cell lines, named TSCs, from fetal thymus. TSCs express some of the markers present on tissue progenitor/stem cells such as Sca-1. Gene expression profiling verifies the thymic identity of TSCs. RANK stimulation of these cells induces expression of autoimmune regulator (Aire) and Aire-dependent tissue-restricted antigens (TRAs) in TSCs in vitro. TSCs could be differentiated into medullary thymic epithelial cell-like cells with exogenously expressed NF-?B subunits RelB and p52. Importantly, upon transplantation under the kidney capsules of nude mice, TSCs are able to differentiate into mature TEC-like cells that can support some limited development of T cells in vivo. These findings suggest that the TSC lines we established bear some characteristics of TEPC cells and are able to differentiate into functional TEC-like cells in vitro and in vivo. The cloned TEPC-like cell lines may provide useful tools to study the differentiation of mature TEC cells from precursors. PMID:24086471

Chen, Pengfei; Zhang, Jun; Zhan, Yu; Su, Juanjuan; Du, Yarui; Xu, Guoliang; Shi, Yufang; Siebenlist, Ulrich; Zhang, Xiaoren

2013-09-23

286

Established Thymic Epithelial Progenitor/Stem Cell-Like Cell Lines Differentiate into Mature Thymic Epithelial Cells and Support T Cell Development  

PubMed Central

Common thymic epithelial progenitor/stem cells (TEPCs) differentiate into cortical and medullary thymic epithelial cells (TECs), which are required for the development and selection of thymocytes. Mature TEC lines have been widely established. However, the establishment of TEPC lines is rarely reported. Here we describe the establishment of thymic epithelial stomal cell lines, named TSCs, from fetal thymus. TSCs express some of the markers present on tissue progenitor/stem cells such as Sca-1. Gene expression profiling verifies the thymic identity of TSCs. RANK stimulation of these cells induces expression of autoimmune regulator (Aire) and Aire-dependent tissue-restricted antigens (TRAs) in TSCs in vitro. TSCs could be differentiated into medullary thymic epithelial cell-like cells with exogenously expressed NF-?B subunits RelB and p52. Importantly, upon transplantation under the kidney capsules of nude mice, TSCs are able to differentiate into mature TEC-like cells that can support some limited development of T cells in vivo. These findings suggest that the TSC lines we established bear some characteristics of TEPC cells and are able to differentiate into functional TEC-like cells in vitro and in vivo. The cloned TEPC-like cell lines may provide useful tools to study the differentiation of mature TEC cells from precursors.

Zhan, Yu; Su, Juanjuan; Du, Yarui; Xu, Guoliang; Shi, Yufang; Siebenlist, Ulrich; Zhang, Xiaoren

2013-01-01

287

Host epithelial geometry regulates breast cancer cell invasiveness.  

PubMed

Breast tumor development is regulated in part by cues from the local microenvironment, including interactions with neighboring nontumor cells as well as the ECM. Studies using homogeneous populations of breast cancer cell lines cultured in 3D ECM have shown that increased ECM stiffness stimulates tumor cell invasion. However, at early stages of breast cancer development, malignant cells are surrounded by normal epithelial cells, which have been shown to exert a tumor-suppressive effect on cocultured cancer cells. Here we explored how the biophysical characteristics of the host microenvironment affect the proliferative and invasive tumor phenotype of the earliest stages of tumor development, by using a 3D microfabrication-based approach to engineer ducts composed of normal mammary epithelial cells that contained a single tumor cell. We found that the phenotype of the tumor cell was dictated by its position in the duct: proliferation and invasion were enhanced at the ends and blocked when the tumor cell was located elsewhere within the tissue. Regions of invasion correlated with high endogenous mechanical stress, as shown by finite element modeling and bead displacement experiments, and modulating the contractility of the host epithelium controlled the subsequent invasion of tumor cells. Combining microcomputed tomographic analysis with finite element modeling suggested that predicted regions of high mechanical stress correspond to regions of tumor formation in vivo. This work suggests that the mechanical tone of nontumorigenic host epithelium directs the phenotype of tumor cells and provides additional insight into the instructive role of the mechanical tumor microenvironment. PMID:23150585

Boghaert, Eline; Gleghorn, Jason P; Lee, KangAe; Gjorevski, Nikolce; Radisky, Derek C; Nelson, Celeste M

2012-11-12

288

Inhibition of HIV-1 Infectivity and Epithelial Cell Transfer by Human Monoclonal IgG and IgA Antibodies Carrying the b12 V Region1  

PubMed Central

Both IgG and secretory IgA Abs in mucosal secretions have been implicated in blocking the earliest events in HIV-1 transit across epithelial barriers, although the mechanisms by which this occurs remain largely unknown. In this study, we report the production and characterization of a human rIgA2 mAb that carries the V regions of IgG1 b12, a potent and broadly neutralizing anti-gp120 Ab which has been shown to protect macaques against vaginal simian/HIV challenge. Monomeric, dimeric, polymeric, and secretory IgA2 derivatives of b12 reacted with gp120 and neutralized CCR5- and CXCR4-tropic strains of HIV-1 in vitro. With respect to the protective effects of these Abs at mucosal surfaces, we demonstrated that IgG1 b12 and IgA2 b12 inhibited the transfer of cell-free HIV-1 from ME-180 cells, a human cervical epithelial cell line, as well as Caco-2 cells, a human colonic epithelial cell line, to human PBMCs. Inhibition of viral transfer was due to the ability of b12 to block both viral attachment to and uptake by epithelial cells. These data demonstrate that IgG and IgA MAbs directed against a highly conserved epitope on gp120 can interfere with the earliest steps in HIV-1 transmission across mucosal surfaces, and reveal a possible mechanism by which b12 protects the vaginal mucosal against viral challenge in vivo.

Mantis, Nicholas J.; Palaia, Jana; Hessell, Ann J.; Mehta, Simren; Zhu, Zhiyi; Corthesy, Blaise; Neutra, Marian R.; Burton, Dennis R.; Janoff, Edward N.

2010-01-01

289

Differentiation of adipose-derived adult stem cells into epithelial-like stem cells.  

PubMed

Adipose-derived adult stem (ADAS) cells can be easily obtained in large quantities. Previous studies have suggested that all-trans-retinoic acid (ATRA) plays an important role in the differentiation of mesenchymal stem cells toward an epithelial lineage. In order to verify that ADAS-cells can differentiate into an epithelial lineage retaining most of the characteristics of stem cells, ADAS-cells were isolated and cultured. They were induced to differentiate toward an epithelial lineage in vitro. Differentiated epithelial cells were assayed as to whether they retain characteristics of stem cells by RT-PCR and cell cycle stage analysis, and were further induced to differentiate toward an osteogenic lineage. RT-PCR analysis revealed that no CK5, CK10 or CK19 mRNA was detected in ADAS-cells, CK19 but not CK5 or CK10 mRNA was detected in differentiated cells at passage 1, CK10 and CK19 expression but not CK5 mRNA was detected in differentiated cells at passage 10. After induction, the expression of CK19 was observed by immunofluorescent staining. Positive staining with alkaline phosphatase (ALP) and Von Kossa staining verified that differentiated epithelial cells still had potential to further differentiate toward an osteogenic lineage. These experiments provide proof that ADAS-cells can differentiate into an epithelial lineage retaining most of the characteristics of stem cells. PMID:23415496

Yan, Yingjun; Liu, Yuxiao; Liu, Daqing; He, Lijuan; Guan, Lidong; Wang, Yunfang; Nan, Xue; Pei, Xuetao

2012-12-07

290

Clinical features of bacterial vaginosis in a murine model of vaginal infection with Gardnerella vaginalis.  

PubMed

Bacterial vaginosis (BV) is a dysbiosis of the vaginal flora characterized by a shift from a Lactobacillus-dominant environment to a polymicrobial mixture including Actinobacteria and gram-negative bacilli. BV is a common vaginal condition in women and is associated with increased risk of sexually transmitted infection and adverse pregnancy outcomes such as preterm birth. Gardnerella vaginalis is one of the most frequently isolated bacterial species in BV. However, there has been much debate in the literature concerning the contribution of G. vaginalis to the etiology of BV, since it is also present in a significant proportion of healthy women. Here we present a new murine vaginal infection model with a clinical isolate of G. vaginalis. Our data demonstrate that this model displays key features used clinically to diagnose BV, including the presence of sialidase activity and exfoliated epithelial cells with adherent bacteria (reminiscent of clue cells). G. vaginalis was capable of ascending uterine infection, which correlated with the degree of vaginal infection and level of vaginal sialidase activity. The host response to G. vaginalis infection was characterized by robust vaginal epithelial cell exfoliation in the absence of histological inflammation. Our analyses of clinical specimens from women with BV revealed a measureable epithelial exfoliation response compared to women with normal flora, a phenotype that, to our knowledge, is measured here for the first time. The results of this study demonstrate that G. vaginalis is sufficient to cause BV phenotypes and suggest that this organism may contribute to BV etiology and associated complications. This is the first time vaginal infection by a BV associated bacterium in an animal has been shown to parallel the human disease with regard to clinical diagnostic features. Future studies with this model should facilitate investigation of important questions regarding BV etiology, pathogenesis and associated complications. PMID:23527214

Gilbert, Nicole M; Lewis, Warren G; Lewis, Amanda L

2013-03-19

291

Clinical Features of Bacterial Vaginosis in a Murine Model of Vaginal Infection with Gardnerella vaginalis  

PubMed Central

Bacterial vaginosis (BV) is a dysbiosis of the vaginal flora characterized by a shift from a Lactobacillus-dominant environment to a polymicrobial mixture including Actinobacteria and Gram-negative bacilli. BV is a common vaginal condition in women and is associated with increased risk of sexually transmitted infection and adverse pregnancy outcomes such as preterm birth. Gardnerella vaginalis is one of the most frequently isolated bacterial species in BV. However, there has been much debate in the literature concerning the contribution of G. vaginalis to the etiology of BV, since it is also present in a significant proportion of healthy women. Here we present a new murine vaginal infection model with a clinical isolate of G. vaginalis. Our data demonstrate that this model displays key features used clinically to diagnose BV, including the presence of sialidase activity and exfoliated epithelial cells with adherent bacteria (reminiscent of clue cells). G. vaginalis was capable of ascending uterine infection, which correlated with the degree of vaginal infection and level of vaginal sialidase activity. The host response to G. vaginalis infection was characterized by robust vaginal epithelial cell exfoliation in the absence of histological inflammation. Our analyses of clinical specimens from women with BV revealed a measureable epithelial exfoliation response compared to women with normal flora, a phenotype that, to our knowledge, is measured here for the first time. The results of this study demonstrate that G. vaginalis is sufficient to cause BV phenotypes and suggest that this organism may contribute to BV etiology and associated complications. This is the first time vaginal infection by a BV associated bacterium in an animal has been shown to parallel the human disease with regard to clinical diagnostic features. Future studies with this model should facilitate investigation of important questions regarding BV etiology, pathogenesis and associated complications.

Gilbert, Nicole M.; Lewis, Warren G.; Lewis, Amanda L.

2013-01-01

292

NOX4-dependent ROS production by stromal mammary cells modulates epithelial MCF-7 cell migration  

PubMed Central

Background: The influence of the stromal microenvironment on the progression of epithelial cancers has been demonstrated. Unravelling the mechanisms by which stromal cells affect epithelial behaviour will contribute in understanding cellular malignancy. It has been proposed that redox environment has a role in the acquisition of malignancy. In this work, we studied the influence of epithelial cells on the stromal redox status and the consequence of this phenomenon on MCF-7 cell motility. Methods: We analysed in a co-culture system, the effect of RMF-EG mammary stromal cells on the migratory capacity of MCF-7 cell line. To test whether the NOX-dependent stromal redox environment influences the epithelial migratory behaviour, we knocked down the expression of NOX4 using siRNA strategy. The effect of TGF-?1 on NOX4 expression and activity was analysed by qPCR, and intracellular ROS production was measured by a fluorescent method. Results: Migration of MCF-7 breast epithelial cells was stimulated when co-cultured with RMF-EG cells. This effect depends on stromal NOX4 expression that, in turn, is enhanced by epithelial soluble factors. Pre-treatment of stromal cells with TGF-?1 enhanced this migratory stimulus by elevating NOX4 expression and intracellular ROS production. TGF-?1 seems to be a major component of the epithelial soluble factors that stimulate NOX4 expression. Conclusions: Our results have identified that an increased stromal oxidative status, mainly provided by an elevated NOX4 expression, is a permissive element in the acquisition of epithelial migratory properties. The capacity of stromal cells to modify their intracellular ROS production, and accordingly, to increase epithelial motility, seems to depend on epithelial soluble factors among which TGF-?1 have a decisive role.

Tobar, N; Guerrero, J; Smith, P C; Martinez, J

2010-01-01

293

Characterisation of cell adhesion in airway epithelial cell types using electric cell-substrate impedance sensing.  

PubMed

Research on epithelial cell lines and primary epithelium is required to dissect the mechanisms underlying the structural abnormalities in airway epithelium observed for respiratory diseases, including asthma and chronic obstructive pulmonary disease. The novel electric cell-substrate impedance sensing technique was used to monitor cell adhesion/spreading, barrier function and wound healing. Primary bronchial epithelium was compared with airway epithelial cell lines 16HBE14o-, BEAS-2B, NCI-H292 and A549. BEAS-2B, A549 and primary cells form a confluent monolayer more rapidly than do 16HBE14o- cells. In contrast, 16HBE14o- cells form stronger intercellular contacts, with a 10-fold higher resistance than BEAS-2B, A549 and NCI-H292 cells and a five-fold increase over primary cells. Accordingly, expression of the adhesion molecules zona occludens-1 and E-cadherin was highest in 16HBE14o- cells. These molecules were localised in intercellular junctions in both 16HBE14o- and primary cells. Finally, restoration of barrier function upon injury was impaired in BEAS-2B compared to 16HBE14o- cells. In conclusion, epithelial cell types display remarkable phenotypic differences and should, accordingly, be used to address specific research questions. 16HBE14o- cells appear most suitable for studies on barrier formation, whereas resemblance in attachment of primary and BEAS-2B and A549 cells makes the latter more important for translational research on cell-matrix contact. PMID:19741028

Heijink, I H; Brandenburg, S M; Noordhoek, J A; Postma, D S; Slebos, D-J; van Oosterhout, A J M

2009-09-09

294

Anthrax Toxin Entry into Polarized Epithelial Cells  

Microsoft Academic Search

applied to the basolateral surface of T84 cells but no response when it was applied to the apical surface. PA alone had no effect when it was applied to either surface. T84 cells exposed basolaterally bound at least 30-fold-more PA than did T84 cells exposed apically, indicating that the PA receptor is largely or completely restricted to the basolateral membrane

KATHRYN E. BEAUREGARD; SUSAN WIMER-MACKIN; R. JOHN COLLIER

1999-01-01

295

Micronucleus investigation in human buccal epithelial cells of gutkha users  

PubMed Central

Background: Gutkha is a cheap and convenient betel quid substitute, which is popular among all age groups. Various studies reveal its carcinogenic nature that leads to oral submucosus fibrosis and increases the chances of oral cancer. The micronucleus (MN) assay in exfoliated mucosal cells is a useful method for observing genetic damage in humans. Aim: To observe the genotoxic effect of gutkha on human buccal epithelial cells. Materials and Methods: The MN assay was performed to assess the frequency of MN in human buccal epithelial cells. The study comprises 60 individuals of which 30 individuals were gutkha chewers and another 30 were nonusers (control). The MN frequency was scored to estimate the genotoxic damage. Results: In gutkha users, the frequency of MN was highly significant (17.4 ± 0.944) as compared with nonusers (control) groups (4.53 ± 0.331) (P < 0.001). Conclusions: The MN assay in human buccal epithelial cells is a useful and minimally invasive method for monitoring genetic damage in humans. Asignificantly higher frequency of micronucleated cells are found among gutkha users.

Jyoti, Smita; Khan, Saif; Afzal, Mohammad; Siddique, Yasir Hasan

2012-01-01

296

Calcium oscillations in gingival epithelial cells infected with Porphyromonas gingivalis.  

PubMed

The periodontal pathogen Porphyromonas gingivalis modulates epithelial cell signal transduction pathways including Ca2+ signaling, and internalizes within the host cell cytoplasm. Since nuclear and cytoplasmic [Ca2+] increases can induce different host cell responses, P. gingivalis-related [Ca2+] changes in these compartments were measured by digital fluorescent imaging microscopy. Non-deconvolved and deconvolved fura-2 images showed that P. gingivalis exposure caused human gingival epithelial cells cultured in physiologic [Ca2+] levels to undergo sustained oscillations of [Ca2+] in nuclear and cytoplasmic spaces. However, P. gingivalis invasion was not tightly correlated with intracellular [Ca2+] oscillations, since invasion could significantly precede, or even occur in the absence of, oscillations. [Ca2+] oscillations required a Ca2+ influx, which was completely inhibited by La3+ or 2-APB (2-aminoethoxydiphenyl borate), indicating Ca2+ entry was via a Ca(2+)-permeable channel. Ca2+ entry was likely not via a store-operated channel, since Ca2+ release from intracellular stores was not observed during cellular uptake of P. gingivalis. Hence, uptake of P. gingivalis in gingival epithelial cells induces oscillations in nuclear and cytoplasmic spaces by activating a Ca2+ influx through Ca2+ channels. PMID:15109958

Belton, Carol M; Goodwin, Paul C; Fatherazi, Sahba; Schubert, Mark M; Lamont, Richard J; Izutsu, Kenneth T

2004-04-01

297

Medullary thymic epithelial cell depletion leads to autoimmune hepatitis.  

PubMed

TRAF6, an E3 ubiquitin protein ligase, plays a critical role in T cell tolerance by regulating medullary thymic epithelial cell (mTEC) development. mTECs regulate T cell tolerance by ectopically expressing self-antigens and eliminating autoreactive T cells in the thymus. Here we show that mice with mTEC depletion due to conditional deletion of Traf6 expression in murine thymic epithelial cells (Traf6?TEC mice) showed a surprisingly narrow spectrum of autoimmunity affecting the liver. The liver inflammation in Traf6?TEC mice exhibited all the histological and immunological characteristics of human autoimmune hepatitis (AIH). The role of T cells in AIH establishment was supported by intrahepatic T cell population changes and AIH development after transfer of liver T cells into immunodeficient mice. Despite a 50% reduction in natural Treg thymic output, peripheral tolerance in Traf6?TEC mice was normal, whereas compensatory T regulatory mechanisms were evident in the liver of these animals. These data indicate that mTECs exert a cell-autonomous role in central T cell tolerance and organ-specific autoimmunity, but play a redundant role in peripheral tolerance. These findings also demonstrate that Traf6?TEC mice are a relevant model with which to study the pathophysiology of AIH, as well as autoantigen-specific T cell responses and regulatory mechanisms underlying this disease. PMID:23867620

Bonito, Anthony J; Aloman, Costica; Fiel, M Isabel; Danzl, Nichole M; Cha, Sungwon; Weinstein, Erica G; Jeong, Seihwan; Choi, Yongwon; Walsh, Matthew C; Alexandropoulos, Konstantina

2013-07-15

298

Role of epithelial-stem cell interactions during dental cell differentiation.  

PubMed

Epithelial-mesenchymal interactions regulate the growth and morphogenesis of ectodermal organs such as teeth. Dental pulp stem cells (DPSCs) are a part of dental mesenchyme, derived from the cranial neural crest, and differentiate into dentin forming odontoblasts. However, the interactions between DPSCs and epithelium have not been clearly elucidated. In this study, we established a mouse dental pulp stem cell line (SP) comprised of enriched side population cells that displayed a multipotent capacity to differentiate into odontogenic, osteogenic, adipogenic, and neurogenic cells. We also analyzed the interactions between SP cells and cells from the rat dental epithelial SF2 line. When cultured with SF2 cells, SP cells differentiated into odontoblasts that expressed dentin sialophosphoprotein. This differentiation was regulated by BMP2 and BMP4, and inhibited by the BMP antagonist Noggin. We also found that mouse iPS cells cultured with mitomycin C-treated SF2-24 cells displayed an epithelial cell-like morphology. Those cells expressed the epithelial cell markers p63 and cytokeratin-14, and the ameloblast markers ameloblastin and enamelin, whereas they did not express the endodermal cell marker Gata6 or mesodermal cell marker brachyury. This is the first report of differentiation of iPS cells into ameloblasts via interactions with dental epithelium. Co-culturing with dental epithelial cells appears to induce stem cell differentiation that favors an odontogenic cell fate, which may be a useful approach for tooth bioengineering strategies. PMID:22298769

Arakaki, Makiko; Ishikawa, Masaki; Nakamura, Takashi; Iwamoto, Tsutomu; Yamada, Aya; Fukumoto, Emiko; Saito, Masahiro; Otsu, Keishi; Harada, Hidemitsu; Yamada, Yoshihiko; Fukumoto, Satoshi

2012-02-01

299

Epithelial differentiation is a determinant in the production of eotaxin-2 and -3 by bronchial epithelial cells in response to IL4 and IL13  

Microsoft Academic Search

The composition of the airway epithelium is dynamic and epithelial differentiation is regulated by endogenous mediators as well as inhaled substances. In atopic asthma the differentiation of the epithelium is altered. Various studies have addressed the ability of cultured airway epithelial cells to release the eosinophil-attractant chemokines eotaxin, eotaxin-2 and eotaxin-3 using epithelial cell lines or poorly differentiated primary cells.

Sandra van Wetering; Suzanne Zuyderduyn; Dennis K. Ninaber; Marianne A. J. A. van Sterkenburg; Klaus F. Rabe; Pieter S. Hiemstra

2007-01-01

300

Overexpression of Smad2 inhibits proliferation of gingival epithelial cells.  

PubMed

BACKGROUND AND OBJECTIVE: Spatiotemporal inhibition of apical migration and proliferation of gingival epithelium are significant factors involved in periodontal regeneration. Transforming growth factor ? (TGF-?) is important in multiple aspects of wound healing, and Smad2, a downstream transcription factor of TGF-?, has an inhibitory effect on re-epithelialization during gingival wound healing. Therefore, we investigated the effects on migration and proliferation status, and intra/extracellular signaling regulated by Smad2 overexpression in gingival epithelial cells. MATERIAL AND METHODS: Gingival epithelial cells were isolated from the palatal gingival tissue of transgenic mice overexpressing Smad2 driven by the Keratin14 promoter. Smad2 expression was identified by western blotting and immunofluorescence analysis. Scratch assay and 5-bromo-2'-deoxyuridine staining were performed to assess cell migration and proliferation. To inactivate TGF-? type I receptor, the cultures were supplemented with SB431542. Secreted TGF-? was quantified by ELISA. Smad2 target gene expression was examined by real-time RT-PCR and in vivo immunofluorescence analysis of gingival junctional epithelium. RESULTS: Smad2-overexpressing cells were confirmed to have significant phosphorylated Smad2 in the nucleus. Scratch assay and 5-bromo-2'-deoxyuridine staining indicated that Smad2-overexpressing cells showed no significant differences in migration, but had reduced proliferation rates compared to wild-type controls. SB431542 significantly inhibited Smad2 phosphorylation, which coincided with restoration of the proliferation rate in Smad2-overexpressing cells. ELISA of TGF-? release did not show any differences between genotypes. The cell cycle inhibitors, p15 and p21, showed significant upregulation in Smad2-overexpressing cells compared to wild-type controls. Moreover, junctional epithelium of the transgenic mice showed increased expression of P-Smad2, p15 and p21. CONCLUSION: The signaling activation triggered by overexpression of Smad2 was dependent on TGF-? type I receptor, and the activated Smad2 increased p15 and p21 expression, responsible for inhibiting cell cycle entry, resulting in antiproliferative effects on gingival epithelial cells. Understanding of Smad2-induced signaling would be useful for possible clinical application to regulate gingival epithelial downgrowth. PMID:23738652

Shimoe, M; Yamamoto, T; Shiomi, N; Tomikawa, K; Hongo, S; Yamashiro, K; Yamaguchi, T; Maeda, H; Takashiba, S

2013-06-01

301

Borrelia burgdorferi bind to epithelial cell proteoglycans.  

PubMed Central

Borrelia burgdorferi adhere to mammalian cells in vitro but neither the ligand(s) nor the receptor(s) has (have) been clearly established. Using an in vitro attachment-inhibition assay, a B. burgdorferi attachment mechanism has been identified. Heparin, heparan sulfate, and dermatan sulfate reduced the attachment of virulent B. burgdorferi strain 297 to HeLa cells by approximately 60%. In addition, virulent, but not avirulent, B. burgdorferi strains B31, N40, and HB19 demonstrated heparin attachment-inhibition. Attachment to Chinese hamster ovary cells deficient in heparan sulfate proteoglycans was reduced by 68% compared to attachment to wild-type cells and was identical to attachment at maximum heparin inhibition to the wild-type cells. Pretreatment of HeLa cell monolayers with heparitinase, heparinase, and chondroitinase ABC, but not with chondroitinase AC, reduced borrelial attachment by approximately 50%. A moderately high affinity, low copy number, promiscuous B. burgdorferi glycosaminoglycan receptor was demonstrated by equilibrium binding studies. A 39-kD polypeptide, purified by heparin affinity chromatography from Triton X-100 extracts derived from virulent borrelia, was a candidate for this receptor. These studies indicate that one mode of B. burgdorferi attachment to eukaryotic cells is mediated by a borrelial glycosaminoglycan receptor attaching to surface-exposed proteoglycans on mammalian cells. Images

Isaacs, R D

1994-01-01

302

Stable cell line of T-SV40 immortalized human glomerular visceral epithelial cells  

Microsoft Academic Search

Stable cell line of T-SV40 immortalized human glomerular visceral epithelial cells. Human subcultures (third passage) of glomerular visceral epithelial cells (VEC) isolated from one month old kidney were successfully transfected by two recombinant plasmids containing the cloned oncogenes from the simian virus 40 large T antigen and H-ras gene. One postcrisis cell clone (56\\/10 A1) was selected, propagated and characterized.

Françoise Delarue; Angela Virone; Jacqueline Hagege; Roger Lacave; Marie-Nëlle Peraldi; Colette Adida; Eric Rondeau; Jean Feunteun; Jean-Daniel Sraer

1991-01-01

303

Cdk5 regulates cell-matrix and cell-cell adhesion in lens epithelial cells.  

PubMed

Cdk5 is a member of the cyclin-dependent kinase family, which is expressed predominantly in terminally differentiated neurons. Lower levels of Cdk5 are also found in a wide variety of cell types, including the lens. Although Cdk5 has been shown to play an important role in neuronal migration and neurite outgrowth, its function in non-neuronal cells is not known. Therefore, this study was undertaken to explore the role of Cdk5 in the lens. Results showed that, within the adult mouse lens, Cdk5 was localized to the cytoplasm, especially along the lateral membranes of differentiating primary fiber cells, which suggests a role in cell-cell adhesion. Staining at the tips of elongating fiber cells was also particularly strong, suggesting a role in cell-matrix adhesion. To examine the possible role of Cdk5 in lens epithelial cell adhesion, we stably transfected N/N1003A rabbit lens epithelial cells with cDNAs for Cdk5 or a dominant-negative mutation, Cdk5-T33. Attachment to a fibronectin matrix, as measured with substrate-coated cell adhesion strips, was increased by Cdk5 overexpression, while an equivalent overexpression of Cdk5-T33 had no effect. Cdk5 also increased the rate of cell attachment and spreading as measured by electric cell-substrate impedance sensing (ECIS). In addition, Cdk5 overexpression decreased cell-cell adhesion as measured by a cell aggregation assay. These findings suggest that Cdk5 plays a role in regulating both cell-matrix and cell-cell interactions in the lens. PMID:11973352

Negash, Sewite; Wang, Hwai-Shi; Gao, Chun; Ledee, Dolena; Zelenka, Peggy

2002-05-15

304

Inflammatory response modulation of airway epithelial cells exposed to formaldehyde.  

PubMed

The two main difficulties when assessing the role and action mechanism of environmental pollutant exposure on the respiratory tract using in vitro methodology are firstly to create exposure conditions that closely mimic the human situation, and secondly to choose an experimental model that accurately represents lung compartment complexity, with different types of cell interaction. The aim of this study was to resolve these two challenges. The first of our difficulties was to find the closest experimental conditions to mimic respiratory environmental pollutant exposure. We compared the effects of formaldehyde (FA) on two cellular models, alveolar and bronchial cell lines, respectively A549 and BEAS-2B. The cells were exposed for 30 min to an environmental dose of gaseous FA (50 ?g/m³) at the air-liquid interface. In order to mimic macrophage-epithelial cell cooperation, sensitizations (with TNF? or with conditioned medium from macrophages--CM) prior to gas exposure were applied. After toxicity evaluation, local inflammation was assessed by IL-8 and MCP-1 production 24h after exposure. In our experimental conditions FA had no effects on alveolar and bronchial epithelial cells without any sensitization. FA exposure after TNF? sensitization alone induced a moderate increase of IL-8 by A549 cells. After sensitization with CM, FA exposure induced a strong increase of IL-8 production by A549 cells in comparison to air, whereas a decrease of MCP-1 production was observed on BEAS-2B cells. It appears that the response of alveolar and bronchial epithelial cells to FA was moderate and that complex sensitization refines the inflammatory response to environmental stresses. When sensitized with CM, these cell lines responded differently to FA exposure. Finally by interacting with the respiratory epithelium, FA could exacerbate the inflammation of airways that occurs in severe asthma, and even synergize the effects of other air pollutants such as allergens. Evaluation of nasal cell inflammatory response could shed further light on the effects of FA on respiratory epithelium. PMID:22484645

Persoz, Charles; Achard, Sophie; Momas, Isabelle; Seta, Nathalie

2012-03-31

305

Stiffness nanotomography of human epithelial cancer cells  

NASA Astrophysics Data System (ADS)

The mechanical stiffness of individual cells is important in both cancer initiation and metastasis. We present atomic force microscopy (AFM) based nanoindentation experiments on various human mammary and esophagus cell lines covering the spectrum from normal immortalized cells to highly metastatic ones. The combination of an AFM with a confocal fluorescence lifetime imaging microscope (FLIM) in conjunction with the ability to move the sample and objective independently allow for precise alignment of AFM probe and laser focus with an accuracy down to a few nanometers. This enables us to correlate the mechanical properties with the point of indentation in the FLIM image. We are using force-volume measurements as well as force indentation curves on distinct points on the cells to compare the elastic moduli of the nuclei, nucleoli, and the cytoplasm, and how they vary within and between individual cells and cell lines. Further, a detailed analysis of the force-indentation curves allows study of the cells' mechanical properties at different indentation depths and to generate 3D elasticity maps.

Staunton, Jack R.; Doss, Bryant L.; Gilbert, C. Michael; Kasas, Sandor; Ros, Robert

2012-02-01

306

CCL20, (gamma)(delta) T cells, and IL-22 in corneal epithelial healing  

Technology Transfer Automated Retrieval System (TEKTRAN)

After corneal epithelial abrasion, leukocytes and platelets rapidly enter the corneal stroma, and CCR6 (+) IL-17(+) gamma delta T cells migrate into the epithelium. Gamma delta T-cell-deficient (TCRd(-/-)) mice have significantly reduced inflammation and epithelial wound healing. Epithelial CCL20 mR...

307

?2-agonists promote host defense against bacterial infection in primary human bronchial epithelial cells  

Microsoft Academic Search

BACKGROUND: Airway epithelial cells are critical in host defense against bacteria including Mycoplasma pneumoniae (Mp) in chronic obstructive pulmonary disease (COPD) and asthma. ?2-agonists are mainstay of COPD and asthma therapy, but whether ?2-agonists directly affect airway epithelial host defense functions is unclear. METHODS: Epithelial cells from bronchial brushings of normal (n = 8), asthma (n = 8) and COPD

Claire A Gross; Russell P Bowler; Rebecca M Green; Andrew R Weinberger; Christina Schnell; Hong Wei Chu

2010-01-01

308

Cultured Alveolar Epithelial Cells From Septic Rats Mimic In Vivo Septic Lung  

Microsoft Academic Search

Sepsis results in the formation of pulmonary edema by increasing in epithelial permeability. Therefore we hypothesized that alveolar epithelial cells isolated from septic animals develop tight junctions with different protein composition and reduced barrier function relative to alveolar epithelial cells from healthy animals. Male rats (200–300g) were sacrificed 24 hours after cecal ligation and double puncture (2CLP) or sham surgery.

Taylor S. Cohen; Gladys Gray Lawrence; Susan S. Margulies

2010-01-01

309

Mesenchymal-to-epithelial transition of intercalating cells in Drosophila renal tubules depends on polarity cues from epithelial neighbours  

PubMed Central

The intercalation of mesenchymal cells into epithelia, through mesenchymal-to-epithelial transition (MET), underlies organogenesis, for example, in nephrogenesis, and tissue regeneration, during cell renewal and wound repair. Despite its importance, surprisingly little is known about the mechanisms that bring about MET in comparison with the related and much-studied, reverse process, epithelial-to-mesenchymal transition (EMT). We analyse the molecular events that regulate MET as stellate cells integrate into the established epithelium of the developing renal tubules in Drosophila. We show that stellate cells polarise as they integrate between epithelial principal cells and that the normal, localised expression of cell polarity proteins in principal cells is required for stellate cells to become epithelial. While the basolateral and apical membranes act as cues for stellate cell polarity, adherens junction integrity is required to regulate their movement through the epithelium; in the absence of these junctions stellate cells continue migrating into the tubule lumen. We also show that expression of basolateral proteins in stellate cells is a prerequisite for their ingression between principal cells. We present a model in which the contacts with successive principal cell membrane domains made by stellate cells as they integrate between them act as a cue for the elaboration of stellate cell polarity. We suggest that the formation of zonula adherens junctions between new cell neighbours establishes their apico-basal positions and stabilises them in the epithelium.

Campbell, Kyra; Casanova, Jordi; Skaer, Helen

2010-01-01

310

[Retinal pigment epithelial cell transplantation: perspective].  

PubMed

Age-related macular degeneration is one of the most serious diseases in elderly people because of its disasterous visual outcome and its prevalence. Even if the submacular and choroidal neovascular membranes could be surgically excised, severe damage or evacuation of retinal pigment epithelium is inevitable in the operated area. Pigmentary dystrophy is also a devastating hereditary eye disease with severe visual disturbance. Up to now, there have been no effective treatments for either of them. We conducted basic experiments on retinal pigment epithelium (RPE) culture, transplantation of the cells to the subretinal space of animals, especially, the Royal College of Surgeon's (RCS) rat, a model of hereditary retinal degeneration, and observed their effects in preventing photoreceptor cell death. 1) We reviewed recent reports of RPE function in relation to cytokine production and autocrine/paracrine function of these ligands. Some cytokines with strong mitogenic effects as nerve trophic/growth factors were able to rescue photoreceptor cell death in dystrophic, ischemic, and light-damaged retinas in the rats. We transplanted allograft pigmented RPE from Long Evans rats or xenograft, human and bovine RPE into the subretinal space of RCS rats, and could observe the retardation of the photoreceptor cell death. 2) As a source of human transplantable RPE in clinical practice, we could use patients' own RPE cells as autografts or those from aborted human fetus eyes as allografts. At present, we cannot use RPE cells from different species as xenografts. We tried to obtain enough RPE cells for culture in vitro from patients with large or giant retinal tears, but were unsuccessful. Cells were easily obtained from fetus eyes, and could be cultured and transplanted as fresh, primary, or multiple passage cells. We also tried cryopreservation of these cells for up to 3 months. Enzymatic expression of tyrosinase, tyrosinase related protein I and II and some other enzymes was examined by proliferating chain reaction to detect possible transformation during the procedure. The cell characteristics were well preserved. In the future, if these RPE cells could be safely kept and available in deep-frozen condition, we could use them clinically at the appropriate time and in appropriate numbers for patients as an "RPE bank" just like an "eye bank" for corneal transplantation. 3) Immunological reaction is very important if we consider this technique for clinical application. Up to now, in experimental animals, no immunological reaction has been reported even for xenograft human RPE in rats, in funduscope and histological examination, because the intraocular space is an immunologically privileged site. But transplantation of human RPE cells with a collagen sheet into the anterior chamber in rabbits caused a definite reaction detected by suppression of the electroretinogram and macrophage infiltration into the subretinal space, not only in the operated eye but also in the contralateral non-operated eye. These results suggest that we must be cautious in clinical use of heterogeneous RPE transplantation. The expression of MHC class II cells was observed in the course of photoreceptor cell degeneration in the RCS rats but it was suppressed if they were rescued by the transplantation of human cultured RPE in these animals. 4) For clinical application of this technique, autografts are naturally much better than the xeno grafts or allografts. We tried to use iris pigment epithelium (IPE) for transplantation because it consists of pigmented cells of neural origin and enough could be obtained with ease by peripheral iridectomy. We also tried transfection of a vector (pCNX2) or vector-inserted cDNA of rat bFGF into the rat IPE and transplanted into the subretinal space of RCS rats. These transfected cells expressed strong mRNA of bFGF. The photoreceptors were well preserved and immunological reaction could not be detected by funduscopical or histological examinat PMID:9022310

Tamai, M

1996-12-01

311

Azithromycin Kills Invasive Aggregatibacter actinomycetemcomitans in Gingival Epithelial Cells  

PubMed Central

Aggregatibacter actinomycetemcomitans invades periodontal pocket epithelium and is therefore difficult to eliminate by periodontal scaling and root planing. It is susceptible to azithromycin, which is taken up by many types of mammalian cells. This led us to hypothesize that azithromycin accumulation by gingival epithelium could enhance the killing of intraepithelial A. actinomycetemcomitans. [3H]azithromycin transport by Smulow-Glickman gingival epithelial cells and SCC-25 oral epithelial cells was characterized. To test our hypothesis, we infected cultured Smulow-Glickman cell monolayers with A. actinomycetemcomitans (Y4 or SUNY 465 strain) for 2 h, treated them with gentamicin to eliminate extracellular bacteria, and then incubated them with azithromycin for 1 to 4 h. Viable intracellular bacteria were released, plated, and enumerated. Azithromycin transport by both cell lines exhibited Michaelis-Menten kinetics and was competitively inhibited by l-carnitine and several other organic cations. Cell incubation in medium containing 5 ?g/ml azithromycin yielded steady-state intracellular concentrations of 144 ?g/ml in SCC-25 cells and 118 ?g/ml in Smulow-Glickman cells. Azithromycin induced dose- and time-dependent intraepithelial killing of both A. actinomycetemcomitans strains. Treatment of infected Smulow-Glickman cells with 0.125 ?g/ml azithromycin killed approximately 29% of the intraepithelial CFU of both strains within 4 h, while treatment with 8 ?g/ml azithromycin killed ?82% of the CFU of both strains (P < 0.05). Addition of carnitine inhibited the killing of intracellular bacteria by azithromycin (P < 0.05). Thus, human gingival epithelial cells actively accumulate azithromycin through a transport system that facilitates the killing of intraepithelial A. actinomycetemcomitans and is shared with organic cations.

Lai, Pin-Chuang

2013-01-01

312

Azithromycin kills invasive Aggregatibacter actinomycetemcomitans in gingival epithelial cells.  

PubMed

Aggregatibacter actinomycetemcomitans invades periodontal pocket epithelium and is therefore difficult to eliminate by periodontal scaling and root planing. It is susceptible to azithromycin, which is taken up by many types of mammalian cells. This led us to hypothesize that azithromycin accumulation by gingival epithelium could enhance the killing of intraepithelial A. actinomycetemcomitans. [(3)H]azithromycin transport by Smulow-Glickman gingival epithelial cells and SCC-25 oral epithelial cells was characterized. To test our hypothesis, we infected cultured Smulow-Glickman cell monolayers with A. actinomycetemcomitans (Y4 or SUNY 465 strain) for 2 h, treated them with gentamicin to eliminate extracellular bacteria, and then incubated them with azithromycin for 1 to 4 h. Viable intracellular bacteria were released, plated, and enumerated. Azithromycin transport by both cell lines exhibited Michaelis-Menten kinetics and was competitively inhibited by l-carnitine and several other organic cations. Cell incubation in medium containing 5 ?g/ml azithromycin yielded steady-state intracellular concentrations of 144 ?g/ml in SCC-25 cells and 118 ?g/ml in Smulow-Glickman cells. Azithromycin induced dose- and time-dependent intraepithelial killing of both A. actinomycetemcomitans strains. Treatment of infected Smulow-Glickman cells with 0.125 ?g/ml azithromycin killed approximately 29% of the intraepithelial CFU of both strains within 4 h, while treatment with 8 ?g/ml azithromycin killed ?82% of the CFU of both strains (P < 0.05). Addition of carnitine inhibited the killing of intracellular bacteria by azithromycin (P < 0.05). Thus, human gingival epithelial cells actively accumulate azithromycin through a transport system that facilitates the killing of intraepithelial A. actinomycetemcomitans and is shared with organic cations. PMID:23274657

Lai, Pin-Chuang; Walters, John D

2012-12-28

313

Selenium deficiency alters epithelial cell morphology and responses to influenza.  

PubMed

It is unknown whether nutritional deficiencies affect the morphology and function of structural cells, such as epithelial cells, and modify the susceptibility to viral infections. We developed an in vitro system of differentiated human bronchial epithelial cells (BEC) grown either under selenium-adequate (Se+) or selenium-deficient (Se-) conditions, to determine whether selenium deficiency impairs host defense responses at the level of the epithelium. Se- BECs had normal SOD activity, but decreased activity of the selenium-dependent enzyme GPX1. Interestingly, catalase activity was also decreased in Se- BECs. Both Se- and Se+ BECs differentiated into a mucociliary epithelium; however, Se- BEC demonstrated increased mucus production and increased Muc5AC mRNA levels. This effect was also seen in Se+ BEC treated with 3-aminotriazole, an inhibitor of catalase activity, suggesting an association between catalase activity and mucus production. Both Se- and Se+ were infected with influenza A/Bangkok/1/79 and examined 24 h postinfection. Influenza-induced IL-6 production was greater while influenza-induced IP-10 production was lower in Se- BECs. In addition, influenza-induced apoptosis was greater in Se- BEC as compared to the Se+ BECs. These data demonstrate that selenium deficiency has a significant impact on the morphology and influenza-induced host defense responses in human airway epithelial cells. PMID:17512462

Jaspers, I; Zhang, W; Brighton, L E; Carson, J L; Styblo, M; Beck, M A

2007-03-24

314

Glutamatergic signaling maintains the epithelial phenotype of proximal tubular cells.  

PubMed

Epithelial-mesenchymal transition (EMT) contributes to the progression of renal tubulointerstitial fibrosis. The N-methyl-d-aspartate receptor (NMDAR), which is present in proximal tubular epithelium, is a glutamate receptor that acts as a calcium channel. Activation of NMDAR induces actin rearrangement in cells of the central nervous system, but whether it helps maintain the epithelial phenotype of the proximal tubule is unknown. Here, knockdown of NMDAR1 in a proximal tubule cell line (HK-2) induced changes in cell morphology, reduced E-cadherin expression, and increased ?-SMA expression. Induction of EMT with TGF-?1 led to downregulation of both E-cadherin and membrane-associated ?-catenin, reorganization of F-actin, expression of mesenchymal markers de novo, upregulation of Snail1, and increased cell migration; co-treatment with NMDA attenuated all of these changes. Furthermore, NMDA reduced TGF-?1-induced phosphorylation of Erk1/2 and Akt and the activation of Ras, suggesting that NMDA antagonizes TGF-?1-induced EMT by inhibiting the Ras-MEK pathway. In the unilateral ureteral obstruction model, treatment with NMDA blunted obstruction-induced upregulation of ?-SMA, FSP1, and collagen I and downregulation of E-cadherin. Taken together, these results suggest that NMDAR plays a critical role in preserving the normal epithelial phenotype and modulating tubular EMT. PMID:21597037

Bozic, Milica; de Rooij, Johan; Parisi, Eva; Ortega, Marta Ruiz; Fernandez, Elvira; Valdivielso, José M

2011-05-19

315

Building Epithelial Tissues from Skin Stem Cells  

PubMed Central

The skin epidermis and its appendages provide a protective barrier that guards against loss of fluids, physical trauma, and invasion by harmful microbes. To perform these functions while confronting the harsh environs of the outside world, our body surface undergoes constant rejuvenation through homeostasis. In addition, it must be primed to repair wounds in response to injury. The adult skin maintains epidermal homeostasis, hair regeneration, and wound repair through the use of its stem cells. What are the properties of skin stem cells, when do they become established during embryogenesis, and how are they able to build tissues with such remarkably distinct architectures? How do stem cells maintain tissue homeostasis and repair wounds and how do they regulate the delicate balance between proliferation and differentiation? What is the relationship between skin cancer and mutations that perturbs the regulation of stem cells? In the past 5 years, the field of skin stem cells has bloomed as we and others have been able to purify and dissect the molecular properties of these tiny reservoirs of goliath potential. We report here progress on these fronts, with emphasis on our laboratory’s contributions to the fascinating world of skin stem cells.

Fuchs, E.; Nowak, J.A.

2009-01-01

316

Vaginal Discharge  

MedlinePLUS

... also be on the lookout for symptoms of yeast infections, bacterial vaginosis and trichomoniasis, 3 infections that ... cause changes in your vaginal discharge. Signs of yeast infections White, cottage cheese-like discharge Swelling and ...

317

Estrogen Vaginal  

MedlinePLUS

... estradiol vaginal ring is also used to treat hot flushes ('hot flashes'; sudden strong feelings of heat and sweating) ... mild soap and warm water. Do not use hot water or boil the applicator. Ask your pharmacist ...

318

Isolation, growth and differentiation of hair cell progenitors from the newborn rat cochlear greater epithelial ridge  

Microsoft Academic Search

Mammalian cochlear hair cell loss is irreversible and leads to permanent hearing loss. To restore hearing physiologically, it is necessary to generate new functional hair cells either from endogenous cells or from exogenously transplanted hair cells\\/progenitors. Previous studies suggest that cochlear greater epithelial ridge (GER) and lesser epithelial ridge (LER) cells are capable of differentiating into hair cells. While it

Yuan Zhang; Suo-qiang Zhai; Jianyong Shou; Wei Song; Jian-he Sun; Wei Guo; Gui-liang Zheng; Yin-yan Hu; Wei-Qiang Gao

2007-01-01

319

Cadherin-23 Mediates Heterotypic Cell-Cell Adhesion between Breast Cancer Epithelial Cells and Fibroblasts  

PubMed Central

In the early stages of breast cancer metastasis, epithelial cells penetrate the basement membrane and invade the surrounding stroma, where they encounter fibroblasts. Paracrine signaling between fibroblasts and epithelial tumor cells contributes to the metastatic cascade, but little is known about the role of adhesive contacts between these two cell types in metastasis. Here we show that MCF-7 breast cancer epithelial cells and normal breast fibroblasts form heterotypic adhesions when grown together in co-culture, as evidenced by adhesion assays. PCR and immunoblotting show that both cell types express multiple members of the cadherin superfamily, including the atypical cadherin, cadherin-23, when grown in isolation and in co-culture. Immunocytochemistry experiments show that cadherin-23 localizes to homotypic adhesions between MCF-7 cells and also to heterotypic adhesions between the epithelial cells and fibroblasts, and antibody inhibition and RNAi experiments show that cadherin-23 plays a role in mediating these adhesive interactions. Finally, we show that cadherin-23 is upregulated in breast cancer tissue samples, and we hypothesize that heterotypic adhesions mediated by this atypical cadherin may play a role in the early stages of metastasis.

Apostolopoulou, Maria; Ligon, Lee

2012-01-01

320

Novel human bronchial epithelial cell lines for cystic fibrosis research  

PubMed Central

Immortalization of human bronchial epithelial (hBE) cells often entails loss of differentiation. Bmi-1 is a protooncogene that maintains stem cells, and its expression creates cell lines that recapitulate normal cell structure and function. We introduced Bmi-1 and the catalytic subunit of telomerase (hTERT) into three non-cystic fibrosis (CF) and three ?F508 homozygous CF primary bronchial cell preparations. This treatment extended cell life span, although not as profoundly as viral oncogenes, and at passages 14 and 15, the new cell lines had a diploid karyotype. Ussing chamber analysis revealed variable transepithelial resistances, ranging from 200 to 1,200 ?·cm2. In the non-CF cell lines, short-circuit currents were stimulated by forskolin and inhibited by CFTR(inh)-172 at levels mostly comparable to early passage primary cells. CF cell lines exhibited no forskolin-stimulated current and minimal CFTR(inh)-172 response. Amiloride-inhibitable and UTP-stimulated currents were present, but at lower and higher amplitudes than in primary cells, respectively. The cells exhibited a pseudostratified morphology, with prominent apical membrane polarization, few apoptotic bodies, numerous mucous secretory cells, and occasional ciliated cells. CF and non-CF cell lines produced similar levels of IL-8 at baseline and equally increased IL-8 secretion in response to IL-1?, TNF-?, and the Toll-like receptor 2 agonist Pam3Cys. Although they have lower growth potential and more fastidious growth requirements than viral oncogene transformed cells, Bmi-1/hTERT airway epithelial cell lines will be useful for several avenues of investigation and will help fill gaps currently hindering CF research and therapeutic development.

Fulcher, M. L.; Gabriel, S. E.; Olsen, J. C.; Tatreau, J. R.; Gentzsch, M.; Livanos, E.; Saavedra, M. T.; Salmon, P.; Randell, S. H.

2009-01-01

321

CFTR and tight junctions in cultured bronchial epithelial cells.  

PubMed

Airway epithelial salt and water transport takes place through paracellular and transcellular pathways. This transport depends critically on the epithelial sodium channel (ENaC) and the cystic fibrosis transmembrane conductance regulator (CFTR), operating in concert with the paracellular pathway through the tight junctions (TJ). Normal (16HBE14o-), cystic fibrosis (CFBE41o-), and corrected CFBE41o- (CFBE41o-pCep4 overexpressing wtCFTR) airway epithelial cell lines were cultured under isotonic conditions. Transepithelial electrical resistance (TEER) was measured as indicator of the tightness of the cultures. Morphology was investigated by immunofluorescence and paracellular permeability by lanthanum nitrate or [14C] mannitol as permeability markers. The CFTR-defective cell line CFBE41o- developed higher TEER than its corrected counterpart CFBE41o-pCep4. Addition of a specific inhibitor of CFTR (CFTR(inh)-172) to 16HBE14o- and CFBE41o-pCep4 cells resulted in a time-dependent increase in TEER, whereas stimulation of CFTR by IBMX and forskolin caused a decrease. Permeability to lanthanum and [14C] mannitol was lower in CFBE41o- and in 16HBE14o- cells exposed to CFTR(inh)-172, compared to untreated 16HBE14o- and CFBE41o-pCep4 cells, respectively. 16HBE14o- cells exposed to IBMX and forskolin showed higher permeability to lanthanum but lower permeability to [14C] mannitol compared to control. Immunofluorescence revealed a disorganization of F-actin and alpha-tubulin in 16HBE14o- cells and CFBE41o- pCep4 exposed to CFTR(inh)-172 and in CFBE41o- cells. Changes in F-actin and alpha-tubulin in 16HBE14o- cells exposed to IBMX and forskolin were also seen. These results suggest the possibility of an interaction between CFTR and the TJ protein complex, probably via the cytoskeleton. PMID:19818767

Nilsson, Harriet E; Dragomir, Anca; Lazorova, Lucia; Johannesson, Marie; Roomans, Godfried M

2009-10-08

322

Roles of Wnt\\/?-catenin signaling in epithelial differentiation of mesenchymal stem cells  

Microsoft Academic Search

Bone marrow-derived mesenchymal stem cells (MSCs) have been demonstrated to be able to differentiate into epithelial lineage, but the precise mechanisms controlling this process are unclear. Our aim is to explore the roles of Wnt\\/?-catenin in the epithelial differentiation of MSCs. Using indirect co-culture of rat MSCs with rat airway epithelial cells (RTE), MSCs expressed several airway epithelial markers (cytokeratin

Yajing Wang; Zhaorui Sun; Xuefeng Qiu; Yan Li; Jizheng Qin; Xiaodong Han

2009-01-01

323

Intestinal epithelial cells and their role in innate mucosal immunity  

Microsoft Academic Search

The mucosal surfaces of the respiratory, gastrointestinal and urogenital tracts are covered by a layer of epithelial cells\\u000a that are responsible for sensing and promoting a host immune response in order to establish the limits not only for commensal\\u000a microorganisms but also for foreign organisms or particles. This is a remarkable task as the human body represents a composite\\u000a of

A. L. Maldonado-Contreras; Beth A. McCormick

2011-01-01

324

Bombesin-like peptide receptors in human bronchial epithelial cells  

Microsoft Academic Search

Northern blot and RNAse protection assays previously failed to detect bombesin-like peptide (BLP) receptors in normal human lung tissue, but by RT\\/PCR cultured human bronchial epithelial (HBE) cells expressed all three BLP receptor subtypes, predominantly neuromedin B (NMB) receptor. By RT\\/PCR, we found expression of all three BLP receptor subtypes by human lung tissue and confirmed NMB receptor expression in

Madeleine A. Kane; Miho Toi-Scott; Gary L. Johnson; Kimberly K. Kelley; Dorothy Boose; Antonio Escobedo-Morse

1996-01-01

325

Ivermectin Inhibits Growth of Chlamydia trachomatis in Epithelial Cells  

PubMed Central

Ivermectin is currently approved for treatment of both clinical and veterinary infections by nematodes, including Onchocerca cervicalis in horses and Onchocerca volvulus in humans. However, ivermectin has never been shown to be effective against bacterial pathogens. Here we show that ivermectin also inhibits infection of epithelial cells by the bacterial pathogen, Chlamydia trachomatis, at doses that could be envisioned clinically for sexually-transmitted or ocular infections by Chlamydia.

Pettengill, Matthew A.; Lam, Verissa W.; Ollawa, Ikechukwu; Marques-da-Silva, Camila; Ojcius, David M.

2012-01-01

326

Respiratory epithelial cells in innate immunity to influenza virus infection  

Microsoft Academic Search

Infection by influenza virus leads to respiratory failure characterized by acute lung injury associated with alveolar edema,\\u000a necrotizing bronchiolitis, and excessive bleeding. Severe reactions to infection that lead to hospitalizations and\\/or death\\u000a are frequently attributed to an exuberant host response, with excessive inflammation and damage to the epithelial cells that\\u000a mediate respiratory gas exchange. The respiratory mucosa serves as a

Catherine J. Sanders; Peter C. Doherty; Paul G. Thomas

2011-01-01

327

Effect of ophthalmic viscosurgical devices on lens epithelial cells  

Microsoft Academic Search

Purpose: To investigate the morphological effects of Viscoat® (sodium hyaluronate 3.0%?chondroitin sulfate 4.0%) on lens epithelial cells (LECs).Setting: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA, and the Laboratory of Ultrastructural Morphology, Zoological Institute, University of Liège, Liège, Belgium.Methods: Human LECs collected via capsulorhexis were examined by light microscopy (LM) and transmission electron microscopy (TEM). Lens

Camille Budo; G Goffinet; Dennis Bellotto; W. Matthew Petroll

2003-01-01

328

Isolation of intestinal epithelial cells and evaluation of transport functions  

SciTech Connect

Epithelial cells can be isolated from the small intestine of chickens by a procedure involving hyaluronidase treatment of the intact tissue. The isolated cells retain a high degree of functional activity as assessed by the formation of 70-fold gradients of alpha-MG. Stability of the sugar gradients reflects maintenance of stable electrochemical Na+ gradients across the plasma membrane. The cells can be used to evaluate the properties of Na(+)-dependent sugar transport, Na(+)-independent sugar transport, ion transport, metabolism, membrane potentials, and the integration of these events, all of which are important to achieving a stable sugar gradient.

Kimmich, G.A.

1990-01-01

329

Practical Guide to Diagnosing and Treating Vaginitis.  

PubMed

Bacterial vaginosis (BV), candidiasis, and trichomoniasis account for more than 90% of vaginal infections. BV typically is associated with a decrease in commensal, protective lactobacilli and a proliferation of other flora. Mobiluncus is pathognomonic but found in only 20% of cases. Presence of 3 of 4 criteria indicates BV: a homogenous noninflammatory discharge (not many WBCs); pH >4.5; clue cells (bacteria attached to borders of epithelial cells, > 20 % of epithelial cells); and a positive whiff test. New intravaginal BV preparations cause less-adverse systemic effects than oral regimens. Trichomonas vaginalis, a protozoan, appears to be sexually transmitted and causes up to 25% of vaginitis cases. Diagnosis is made by observation of a foul, frothy discharge; pH >4.5 (present in 70% of cases); punctate cervical microhemorrhages (25% of cases); and motile trichomonads on wet mount (50%-75% of cases). Recommended treatment is a single 2g dose of oral metronidazole. Treatment failure is usually due to nontreatment of the male partner. Candidiasis typically presents as a thick, "curdled" white discharge or vulvar pruritus, with a hyperemic vagina and an erythematous and/or excoriated vulva. Vaginal pH is usually in the normal range of 3.8-4.2 in uncomplicated candidiasis. Microscopic examination of the discharge reveals hyphae or budding yeast in 50%-70% of cases. While the most common offender is Candida albicans, Candida tropicalis and Candida glabrata have become increasingly prevalent. Approximately 15% of C albicans organisms are resistant to clotrimazole and miconazole. Recurrent infections may be treated with fluconazole 150mg weekly for up to 12 consecutive weeks. PMID:9746676

Plourd

1997-02-01

330

Airway epithelial cell tolerance to Pseudomonas aeruginosa  

Microsoft Academic Search

BACKGROUND: The respiratory tract epithelium is a critical environmental interface that regulates inflammation. In chronic infectious airway diseases, pathogens may permanently colonize normally sterile luminal environments. Host-pathogen interactions determine the intensity of inflammation and thus, rates of tissue injury. Although many cells become refractory to stimulation by pathogen products, it is unknown whether the airway epithelium becomes either tolerant or

Qi Wu; Zhong Lu; Margrith W Verghese; Scott H Randell

2005-01-01

331

Comparative study of the dynamics of focal contacts in live epithelial and mesenchymal cells.  

PubMed

To further characterize the morphology and dynamics of focal contacts (FCs) in epithelial cells, we compared the size, number, localization, velocity, and turnover of FCs in epithelial and mesenchymal cell lines. Using immunocytochemistry, we found there were no significant differences between mesenchymal and epithelial cells in number and appearance whereas the location and size of FCs in each cell were different between mesenchymal and epithelial cells. FCs in mesenchymal cells localized at the cell periphery and cell center, but FCs were found only at the cell periphery in epithelial cells. The size of FCs in epithelial cells were significantly smaller than in mesenchymal cells. Next, we compared the dynamics of FCs in both mesenchymal and epithelial cells and found no significant difference between the two groups. Finally, we added inhibitors for the hemidesmosome (HD) proteins, ?6 integrin and ?4 integrin, to HaCat cell (epithelial) cultures and examined the number and size of FCs. Under these conditions, the size and localization of FCs in HaCat cells became comparable to that of mesenchymal cells. Therefore, we concluded the size and localization of FCs is regulated by the existence of HDs in epithelial cells. PMID:21424934

Ozawa, Toshiyuki; Tsuruta, Daisuke

2011-03-23

332

Tissue-regenerating, vision-restoring corneal epithelial stem cells.  

PubMed

The cornea, the most anterior segment of the eye, provides us with exquisite vision. Unlike other vital tissues, it is poorly protected from the environment and is thus reliant on a self-renewal program to preserve integrity. This function is reserved for corneal epithelial stem cells located in the basal layer of the limbus, a narrow transition zone that segregates the peripheral cornea from the adjacent conjunctiva. Under physiological conditions, these cells replenish the corneal epithelium when mature or traumatized cells are lost. However, when the limbus is extensively damaged, stem cell activity is compromised, resulting in a condition known as limbal stem cell deficiency (LSCD). This disease is characterized by corneal neovascularization and persistent epithelial defects which impair vision. Over the past 20 years a myriad of treatment options have been developed for LSCD, most of which incorporate stem cell transplantation. Due to the disadvantages associated with the use of allogeneic and xenogeneic material, researchers are currently focusing on refining techniques involving autologous limbal tissue transplantation and are delving into the possibility that stem cells found in other organs can provide an alternative source of corneal epithelium. Determining where donor stem cells reside on the recipient's ocular surface and how long they remain viable will provide further insights into improving current therapeutic options for patients with LSCD. PMID:21069583

Echevarria, Timothy Jerome; Di Girolamo, Nick

2011-06-01

333

p63 regulates an adhesion programme and cell survival in epithelial cells  

Microsoft Academic Search

p63 is critical for epithelial development yet little is known about the transcriptional programmes it regulates. By characterising transcriptional changes and cellular effects following modulation of p63 expression, we have defined a vital role for p63 in cellular adhesion. Knockdown of p63 expression caused downregulation of cell adhesion-associated genes, cell detachment and anoikis in mammary epithelial cells and keratinocytes. Conversely,

Danielle K. Carroll; Jason S. Carroll; Chee-Onn Leong; Fang Cheng; Myles Brown; Alea. A. Mills; Leif W. Ellisen; Joan S. Brugge

2006-01-01

334

Airway epithelial cells from asthmatic children differentially express proremodeling factors  

PubMed Central

Background The airway epithelium can express factors that drive subepithelial airway remodeling. TGF-?2, vascular epithelial growth factor (VEGF), a disintegrin and metalloprotease 33 (ADAM33), and periostin are hypothesized to be involved in subepithelial remodeling and are overexpressed in adult asthmatic airways. Epidemiologic data suggest that lung function deficits in asthmatic patients are acquired in childhood. Objectives We sought to determine whether airway epithelial cells (AECs) from asthmatic children differentially express TGF-?2, VEGF, ADAM33, or periostin compared with cells from atopic nonasthmatic and healthy children intrinsically or in response to IL-4/IL-13 stimulation. Methods Bronchial and nasal epithelial cells were obtained from brushings from well-characterized asthmatic (n = 16), atopic nonasthmatic (n = 9), and healthy (n = 15) children after achievement of anesthesia for elective procedures. After differentiation at an air-liquid interface (ALI) for 3 weeks, conditioned media were sampled and RNA was extracted from unstimulated and IL-4/IL-13–stimulated cultures. TGF-?2 and VEGF levels were measured with ELISA. ADAM33 and periostin expression was assessed by using real-time PCR. Results TGF-?2 and VEGF production was significantly greater in bronchial and nasal ALI cultures from asthmatic children than in cultures from atopic nonasthmatic and healthy children. TGF-?2 levels increased significantly in asthmatic cultures after IL-4/IL-13 stimulation. Within-subject correlation between nasal and bronchial ALI production of TGF-?2 (r = 0.64, P = .001) and VEGF (r = 0.73, P < .001) was good. Periostin expression was 3.7-fold higher in bronchial cells (P < .001) and 3.9-fold higher in nasal cells (P < .004) from asthmatic children than in cells from atopic nonasthmatic or healthy children. ADAM33 was not differentially expressed by AECs from asthmatic patients compared with that from cells from atopic nonasthmatic or healthy children. Conclusion AECs from asthmatic children differentially express TGF-?2, VEGF, and periostin compared with cells from atopic nonasthmatic and healthy children. Nasal epithelial cells might be a suitable surrogate for bronchial cells that could facilitate investigation of the airway epithelium in future longitudinal pediatric studies.

Lopez-Guisa, Jesus M.; Powers, Claire; File, Daniele; Cochrane, Elizabeth; Jimenez, Nathalia; Debley, Jason S.

2012-01-01

335

Prion infection of epithelial Rov cells is a polarized event.  

PubMed

During prion infections, the cellular glycosylphosphatidylinositol-anchored glycoprotein PrP is converted into a conformational isoform. This abnormal conformer is thought to recruit and convert the normal cellular PrP into a likeness of itself and is proposed to be the infectious agent. We investigated the distribution of the PrP protein on the surface of Rov cells, an epithelial cell line highly permissive to prion multiplication, and we found that PrP is primarily expressed on the apical side. We further show that prion transmission to Rov cells is much more efficient if infectivity contacts the apical side, indicating that the apical and basolateral sides of Rov cells are not equally competent for prion infection and adding prions to the list of the conventional infectious agents (viruses and bacteria) that infect epithelial cells in a polarized manner. These data raise the possibility that apically expressed PrP may be involved in this polarized process of infection. This would add further support for a crucial role of PrP at the cell surface in prion infection of target cells. PMID:15194791

Paquet, Sophie; Sabuncu, Elifsu; Delaunay, Jean-Louis; Laude, Hubert; Vilette, Didier

2004-07-01

336

Rho GTPases and Regulation of Cell Migration and Polarization in Human Corneal Epithelial Cells  

PubMed Central

Purpose Epithelial cell migration is required for regeneration of tissues and can be defective in a number of ocular surface diseases. This study aimed to determine the expression pattern of Rho family small G-proteins in human corneal epithelial cells to test their requirement in directional cell migration. Methods Rho family small G-protein expression was assessed by reverse transcription-polymerase chain reaction. Dominant-inhibitory constructs encoding Rho proteins or Rho protein targeting small interfering RNA were transfected into human corneal epithelial large T antigen cells, and wound closure rate were evaluated by scratch wounding assay, and a complementary non-traumatic cell migration assay. Immunofluorescence staining was performed to study cell polarization and to assess Cdc42 downstream effector. Results Cdc42, Chp, Rac1, RhoA, TC10 and TCL were expressed in human corneal epithelial cells. Among them, Cdc42 and TCL were found to significantly affect cell migration in monolayer scratch assays. These results were confirmed through the use of validated siRNAs directed to Cdc42 and TCL. Scramble siRNA transfected cells had high percentage of polarized cells than Cdc42 or TCL siRNA transfected cells at the wound edge. We showed that the Cdc42-specific effector p21-activated kinase 4 localized predominantly to cell-cell junctions in cell monolayers, but failed to translocate to the leading edge in Cdc42 siRNA transfected cells after monolayer wounding. Conclusion Rho proteins expressed in cultured human corneal epithelial cells, and Cdc42, TCL facilitate two-dimensional cell migration in-vitro. Although silencing of Cdc42 and TCL did not noticeably affect the appearance of cell adhesions at the leading edge, the slower migration of these cells indicates both GTP-binding proteins play important roles in promoting cell movement of human corneal epithelial cells.

Hou, Aihua; Toh, Li Xian; Gan, Kah Hui; Lee, Khee Jin Ryan; Manser, Edward; Tong, Louis

2013-01-01

337

Epithelial Cell Polarity Affects Susceptibility to Pseudomonas aeruginosa Invasion and Cytotoxicity  

Microsoft Academic Search

Intact tissues are relatively resistant to Pseudomonas aeruginosa-induced disease, and injury predisposes tissue to infection. Intact epithelia contain polarized cells that have distinct apical and basolateral membranes with unique lipids and proteins. In this study, the role of cell polarity in epithelial cell susceptibility to P. aeruginosa virulence mechanisms was tested. Madin-Darby canine kidney (MDCK) cells, human corneal epithelial cells,

SUZANNE M. J. FLEISZIG; DAVID J. EVANS; NINA DO; VICKY VALLAS; SUSAN SHIN; KEITH E. MOSTOV

1997-01-01

338

Reconstitution of Mammary Epithelial Morphogenesis by Murine Embryonic Stem Cells Undergoing Hematopoietic Stem Cell Differentiation  

Microsoft Academic Search

BackgroundMammary stem cells are maintained within specific microenvironments and recruited throughout lifetime to reconstitute de novo the mammary gland. Mammary stem cells have been isolated through the identification of specific cell surface markers and in vivo transplantation into cleared mammary fat pads. Accumulating evidence showed that during the reformation of mammary stem cell niches by dispersed epithelial cells in the

Shuxian Jiang; Byeong-Chel Lee; Yigong Fu; Shalom Avraham; Bing Lim; Hava Karsenty Avraham

2010-01-01

339

Oral epithelial cells are susceptible to cell-free and cell-associated HIV-1 infection in vitro.  

PubMed

Epithelial cells lining the oral cavity are exposed to HIV-1 through breast-feeding and oral-genital contact. Genital secretions and breast milk of HIV-1-infected subjects contain both cell-free and cell-associated virus. To determine if oral epithelial cells can be infected with HIV-1 we exposed gingival keratinocytes and adenoid epithelial cells to cell-free virus and HIV-1-infected peripheral blood mononuclear cells and monocytes. Using primary isolates we determined that gingival keratinocytes are susceptible to HIV-1 infection via cell-free CD4-independent infection only. R5 but not X4 viral strains were capable of infecting the keratinocytes. Further, infected cells were able to release infectious virus. In addition, primary epithelial cells isolated from adenoids were also susceptible to infection; both cell-free and cell-associated virus infected these cells. These data have potential implications in the transmission of HIV-1 in the oral cavity. PMID:12954203

Moore, Jennifer S; Rahemtulla, Firoz; Kent, Leigh W; Hall, Stacy D; Ikizler, Mine R; Wright, Peter F; Nguyen, Huan H; Jackson, Susan

2003-09-01

340

Cooperation between epithelial cells demonstrated by potassium transfer  

SciTech Connect

Junction-mediated communication can be measured in fibroblast cultures by determining the ability of mixed cultures of cells sensitive and resistant to ouabain to concentrate K+ in the presence of ouabain. We now report the extension of this assay procedure to cultured epithelial cells. Hamster kidney (HaK) cells maintain their ability to concentrate K+ in ouabain at levels inhibitory to dog kidney (MDCK) cells. When HaK and MDCK cells were cultured together in ouabain-containing medium, the K+ (measured as 86Rb+) in the mixed population was greater than expected if the cells were not interacting. The degree of enhancement, expressed as index of cooperation, depended on the numbers of cells in the cultures, their opportunity for cell-to-cell contact, and (above a certain permissive level) the concentration of ouabain. As with other cell types, protein synthesis in MDCK cells depends on maintenance of cell K+. Autoradiography of cells incubated with (3H)leucine demonstrated that MDCK cells in ouabain-treated mixed cultures were able to synthesize proteins only when physically adjacent to HaK cells. The transmission of labeled nucleosides among the cells provides independent evidence of the phenomenon of cooperation, probably mediated by gap junctions. This system offers promise for investigation of stimuli modulating junctional communication.

Ledbetter, M.L.; Young, G.J.; Wright, E.R.

1986-02-01

341

Angiotensin-converting enzyme in epithelial and neuroepithelial cells.  

PubMed

Angiotensin-converting enzyme (CE) occurs in three types of cell: endothelial, epithelial, and neuroepithelial. In all three, it appears to be bound to plasma membrane. With antisera to the human enzyme, CE is demonstrated in paraffin sections on the apical surface of epithelial cells in the proximal tubule of the kidney, the mucosa of the small intestine, the syncytial trophoblast of the placenta, and the choroid plexus. Epithelial CE is characteristically found on microvillous surfaces in contact with an effluent, well placed to act on substrate in flux. In the brain, CE occurs in nerve fibers and terminals, mainly mesiobasally and in basal ganglia. Mesiobasal CE coincides with other components of the renin-angiotensin system (RAS) in the choroid/ventricular fluid, the subfornical organ, and the magnocellular neurosecretory system of the hypothalamus. Extrapyramidal CE, however, may not be related to the RAS. In the substantia nigra and the globus pallidus, the enzyme has the same cellular distribution as two putative neuromodulators, substance P and enkephalin, the latter a known substrate of CE. PMID:6310427

Defendini, R; Zimmerman, E A; Weare, J A; Alhenc-Gelas, F; Erdös, E G

1983-07-01

342

Proteomic profiling of acrolein adducts in human lung epithelial cells  

PubMed Central

Acrolein (2,3-propenal) is a major indoor and outdoor air pollutant originating largely from tobacco smoke or organic combustion. Given its high reactivity, the adverse effects of inhaled acrolein are likely due to direct interactions with the airway epithelium, resulting in altered epithelial function, but only limited information exists to date regarding the primary direct cellular targets for acrolein. Here, we describe a global proteomics approach to characterize the spectrum of airway epithelial protein targets for Michael adduction in acrolein-exposed bronchial epithelial (HBE1) cells, based on biotin hydrazide labeling and avidin purification of biotinylated proteins or peptides for analysis by LC-MS/MS. Identified protein targets included a number of stress proteins, cytoskeletal proteins, and several key proteins involved in redox signaling, including thioredoxin reductase, thioredoxin, peroxiredoxins, and glutathione S-transferase ?. Because of the central role of thioredoxin reductase in cellular redox regulation, additional LC-MS/MS characterization was performed on purified mitochondrial thioredoxin reductase to identify the specific site of acrolein adduction, revealing the catalytic selenocysteine residue as the target responsible for enzyme inactivation. Our findings indicate that these approaches are useful in characterizing major protein targets for acrolein, and will enhance mechanistic understanding of the impact of acrolein on cell biology.

Spiess, Page C.; Deng, Bin; Hondal, Robert J.; Matthews, Dwight E.; van der Vliet, Albert

2011-01-01

343

Cytotoxicity of Mouthrinses on Epithelial Cells by Micronucleus Test  

PubMed Central

Objectives To determine the cytotoxicity of three commercial mouthrinses Klorhex, Andorex and Tanflex on buccal epithelial cells using micronucleus (MN) test. Materials and Methods 28 patients with aged 16–24 undergone three mouthrinses’ application were analyzed before and after one week exposure. Physiologic saline was used for the control group. The MN incidence was scored in the buccal epithelial of each participants. The difference in pre- and post-treatment after one week incidence of MN and plaque (PI) and gingival indices (GI) was compared by non-parametric statistical tests. Results The micronuclei incidence increased in Klorhex, Tanflex and Andorex groups after exposure to mouth rinses (P<.05). But when compared with the control group, there was not any difference between Andorex and control group (P>.05). In the other study groups, MN incidence was significantly increased after 7 days treatment (P<.05). GI scores of all groups were decreased significantly (P<.05). PI scores were decreased only in the Klorhex group (P<.05). Conclusions Our primary findings support the presence of possible cytotoxic effects of the mouthrinses on gingival epithelial cells.

Erdemir, Ebru Olgun; Sengun, Abdulkadir; Ulker, Mustafa

2007-01-01

344

Redistribution of membrane proteins in isolated mouse intestinal epithelial cells  

PubMed Central

Single mouse intestinal epithelial cells (IEC) may be isolated by the use of a combination of methods used for the isolation of IEC from other species. Isolated cells remain viable for several hours. The membrane integral enzymes alkaline phosphatase and leucine aminopeptidase of isolated IEC are localized to the brush borders of IEC in tissue and in most newly isolated IEC. With time, both enzymes are found distributed over the entire cell surface. Redistribution appears to occur by diffusion in the plane of the membrane. It is slowed, but not blocked, if cells are maintained at 0 degrees C instead of at 37 degrees C, and it is not blocked by fixation in 0.5-3% paraformaldehyde. Drugs that alter cell membrane potential or that affect cell levels of ATP enhance the rate of redistribution of the enzymes.

1980-01-01

345

Nucleostemin as a possible progenitor marker of corneal epithelial cells  

PubMed Central

Purpose Nucleostemin, a nuclear protein involved in the regulation of cell cycle and proliferation, is a candidate marker for various stem cells. We examined the expression of nucleostemin as a marker of differentiation and senescence in corneal epithelial progenitor cells. Methods Nucleostemin expression in normal mouse corneal epithelium was examined by RT-PCR, as well as in cultured mouse corneal epithelial cells by immunohistochemistry. Co-expression with the progenitor markers p63 and Bmi-1, cytokeratin 14 and 19, the proliferating marker Ki67, differentiation markers cytokeratin 12 and involucrin, and with the senescent marker SA-?-gal were examined by immunohistochemistry. Correlation of cellular size and nucleostemin expression was also examined. Results Nucleostemin expression was detected in mouse corneal epithelium by RT-PCR. Immunohistochemistry revealed that nucleostemin was expressed predominantly in basal and suprabasal cells of the whole cornea. The expression of nucleostemin was not associated with the expression of Ki67, K14, and K19, but with the expression of Bmi-1 and particularly with p63. Nucleostemin was not co-expressed with SA-?-gal in same cell. Conclusions Nucleostemin can be used as a progenitor marker analogous to p63.

Kawashima, Motoko; Yoshida, Satoru; Shimmura, Shigeto; Tsubota, Kazuo

2009-01-01

346

Lens epithelial cell proliferation, migration, and metaplasia following capsulorhexis  

PubMed Central

AIM—To study the behaviour of residual lens epithelial cells after capsulorhexis and expression of material from the isolated lens.?METHODS—Human and bovine lens capsules were isolated, sterile non-toxic silicone rings inserted, and the preparations placed in organ culture for up to 12 weeks. Cell coverage of the posterior lens capsule was recorded and the capsules were examined, both pre- and post-coverage, for (a) proliferative activity and (b) cytoskeletal components.?RESULTS—After a lag period outgrowth was observed across the posterior capsule. The rate of cell coverage was dependent upon both species and the presence or absence of serum. The proliferative activity of the cells was greatest at or near the leading edge and decreased once covered. Wrinkles became visible in the posterior capsule during the late stages of pre-coverage and increased in both number and complexity. All cells on both the human and bovine posterior capsules contained F-actin and vimentin and the majority were immunolabelled for ?-smooth muscle actin (?-SMA).?CONCLUSIONS—This model exhibits many of the in vivo characteristics of the lens capsule after extracapsular surgery and may prove useful in further elucidating the cellular mechanisms of posterior capsule opacification.?? Keywords: lens epithelial cells; posterior capsule opacification; capsulorrhexis

Saxby, L.; Rosen, E.; Boulton, M.

1998-01-01

347

Efficient Cultivation Conditions for Human Limbal Epithelial Cells  

PubMed Central

To compare the stem niche in different culture conditions of limbal epithelial cells, the suspended human limbal epithelial cells (HLECs) were seeded on the 3T3-pretreated plates and the other suspended cells were plated on amniotic membranes (AMs) which were either cryo-preserved or freeze-dried. All were cultured for 10 to 12 days. Reverse transcription-polymerase chain reaction (RT-PCR) for ATP-binding casette, subfamily G, member 2 (ABCG2), p63, cytokeratin 12, and connexin 43 were performed in cultivated HLECs and their expression levels were compared. The mRNA expression of all markers examined showed no statistically significant differences between the cells on cryo-preserved and on freeze-dried AM. The expression of p63 and cytokeratin 12 in cultivated cells on AMs were significantly lower than those in 3T3-cocultured cells on RT-PCR and immunofluorescent staining. Cultivated HLECs on AMs showed reduced proliferation and differentiation while maintaining stem-property regardless of the preservative method of AM.

Kim, Mee Kum; Lee, Jae Lim; Oh, Joo Youn; Shin, Mi Sun; Shin, Kyeong Seon; Lee, Jin Hak; Park, Ki Sook; Son, Young Sook

2008-01-01

348

ATP7B detoxifies silver in ciliated airway epithelial cells  

SciTech Connect

Silver is a centuries-old antibiotic agent currently used to treat infected burns. The sensitivity of a wide range of drug-resistant microorganisms to silver killing suggests that it may be useful for treating refractory lung infections. Toward this goal, we previously developed a methylated caffeine silver acetate compound, SCC1, that exhibits broad-spectrum antimicrobial activity against clinical strains of bacteria in vitro and when nebulized to lungs in mouse infection models. Preclinical testing of high concentrations of SCC1 in primary culture mouse tracheal epithelial cells (mTEC) showed selective ciliated cell death. Ciliated cell death was induced by both silver- and copper-containing compounds but not by the methylated caffeine portion of SCC1. We hypothesized that copper transporting P-type ATPases, ATP7A and ATP7B, play a role in silver detoxification in the airway. In mTEC, ATP7A was expressed in non-ciliated cells, whereas ATP7B was expressed only in ciliated cells. The exposure of mTEC to SCC1 induced the trafficking of ATP7B, but not ATP7A, suggesting the presence of a cell-specific silver uptake and detoxification mechanisms. Indeed, the expression of the copper uptake protein CTR1 was also restricted to ciliated cells. A role of ATP7B in silver detoxification was further substantiated when treatment of SCC1 significantly increased cell death in ATP7B shRNA-treated HepG2 cells. In addition, mTEC from ATP7B{sup -/-} mice showed enhanced loss of ciliated cells compared to wild type. These studies are the first to demonstrate a cell type-specific expression of the Ag{sup +}/Cu{sup +} transporters ATP7A, ATP7B, and CTR1 in airway epithelial cells and a role for ATP7B in detoxification of these metals in the lung.

Ibricevic, Aida, E-mail: aidaibricevic@hotmail.co [Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110 (United States); Brody, Steven L., E-mail: sbrody@dom.wustl.ed [Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110 (United States); Youngs, Wiley J., E-mail: youngs@uakron.ed [Department of Chemistry, University of Akron, Akron, OH 44325 (United States); Cannon, Carolyn L., E-mail: carolyn.cannon@utsouthwestern.ed [Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110 (United States)

2010-03-15

349

ATP7B detoxifies silver in ciliated airway epithelial cells  

PubMed Central

Silver is a centuries old antibiotic agent currently used to treat infected burns. The sensitivity of a wide range of drug-resistant microorganisms to silver killing suggests that it may be useful for treating refractory lung infections. Toward this goal, we previously developed a methylated caffeine silver acetate compound, SCC1, that exhibits broad-spectrum antimicrobial activity against clinical strains of bacteria in vitro and when nebulized to lungs in mouse infection models. Preclinical testing of high concentrations of SCC1 in primary culture mouse tracheal epithelial cells (mTEC) showed selective ciliated cell death. Ciliated cell death was induced by both silver- and copper-containing compounds, but not by the methylated caffeine portion of SCC1. We hypothesized that copper transporting P-type ATPases, ATP7A and ATP7B, play a role in silver detoxification in the airway. In mTEC, ATP7A was expressed in non-ciliated cells, whereas ATP7B was expressed only in ciliated cells. The exposure of mTEC to SCC1 induced the trafficking of ATP7B, but not ATP7A, suggesting the presence of a cell-specific silver uptake and detoxification mechanisms. Indeed, the expression of the copper uptake protein CTR1 was also restricted to ciliated cells. A role of ATP7B in silver detoxification was further substantiated when treatment of SCC1 significantly increased cell death in ATP7B shRNA treated HepG2 cells. Additionally, mTEC from ATP7B-/- mice showed enhanced loss of ciliated cells compared to wild type. These studies are the first to demonstrate a cell-type specific expression of the Ag+/Cu+ transporters ATP7A, ATP7B and CTR1 in airway epithelial cells, and a role for ATP7B in detoxification of these metals in the lung.

Ibricevic, Aida; Brody, Steven L.; Youngs, Wiley J.; Cannon, Carolyn L.

2010-01-01

350

Survival differences exhibited by normal and transformed rat liver epithelial cell lines in the aggregate form.  

PubMed

Normal and transformed rat liver epithelial cell lines exhibited differences in the ability to survive in the aggregate form. Normal rat liver epithelial cells in the aggregate form underwent a rapid decline in the number of viable cells, while counterpart transformed epithelial cells exhibited an ability to survive and proliferate in the aggregate form. This survival ability was found to correlate with colony formation in soft agar and tumorigenicity in nude mice. Cell survival in the aggregate form could possibly serve as a criterion for in vitro transformation of epithelial cells derived from rat liver. PMID:870190

Steuer, A F; Hentosh, P M; Diamond, L; Ting, R C

1977-06-01

351

ASBESTOS-INDUCED ACTIVATION OF CELL SIGNALING PATHWAYS IN HUMAN BRONCHIAL EPITHELIAL CELLS  

EPA Science Inventory

Using respiratory epithelial cells transfected with either superoxide dismutase (SOD) or catalase, the authors tested the hypothesis that the activation of the epidermal growth factor (EGF) receptor signal pathway after asbestos exposure involves an oxidative stress. Western blot...

352

Epithelial cells from smokers modify dendritic cell responses in the context of influenza infection.  

EPA Science Inventory

Epidemiologic evidence suggests that cigarette smoking is a risk factor for infection with influenza, but the mechanisms underlying this susceptibility remain unknown. To ascertain if airway epithelial cells from smokers demonstrate a decreased ability to orchestrate an influenza...

353

Rhinovirus infection induces cytotoxicity and delays wound healing in bronchial epithelial cells  

Microsoft Academic Search

BACKGROUND: Human rhinoviruses (RV), the most common triggers of acute asthma exacerbations, are considered not cytotoxic to the bronchial epithelium. Recent observations, however, have questioned this knowledge. The aim of this study was to evaluate the ability of RV to induce epithelial cytotoxicity and affect epithelial repair in-vitro. METHODS: Monolayers of BEAS-2B bronchial epithelial cells, seeded at different densities were

Apostolos Bossios; Stelios Psarras; Dimitrios Gourgiotis; Chrysanthi L Skevaki; Andreas G Constantopoulos; Photini Saxoni-Papageorgiou; Nikolaos G Papadopoulos

2005-01-01

354

Involvement of Fas in regression of vaginal epithelia after ovariectomy and during an estrous cycle.  

PubMed Central

Fas, also called APO-1, belongs to the tumor necrosis factor/nerve growth factor receptor family and transmits an apoptotic signal within the cell by binding to the Fas ligand. Fas has been implicated in the activation-induced suicide of T cells and cytotoxic T cell activity in the immune system. Non-immune cells such as those in liver, lung and ovary also express Fas, but its role in these cells remains unclear. Ovariectomy has been used to study homeostasis of female reproductive organs, which is regulated by sex hormones. Here we analyzed Fas function in the ovariectomy-induced regression of mouse vaginal epithelial cells. Fas expression was detected in vagina and was elevated after ovariectomy. Fas-deficient lpr and lpr(cg) mice did not exhibit ovariectomy-induced regression of vaginal epithelia, whereas uterine regression induced by ovariectomy was not affected in these mice. The vaginas of lpr and lpr(cg) mice were in a persistent estrous stage with cornification of vaginal epithelia, as judged from the cell types in the vaginal fluid. Thus, Fas appears to be involved directly in the regression of vaginal epithelia induced by ovariectomy and during the estrous cycle, suggesting that the physiological role of this receptor extends beyond that exerted on immune cells. This is the first evidence of a role for Fas inducing physiological apoptosis in non-immune cells. Images

Suzuki, A; Enari, M; Eguchi, Y; Matsuzawa, A; Nagata, S; Tsujimoto, Y; Iguchi, T

1996-01-01

355

Helicobacter pylori Activates Gastric Epithelial Cells to Produce Interleukin8 via Protease-Activated Receptor 2  

Microsoft Academic Search

Background\\/Aim: Recently, it has been shown that serine proteases derived from microorganisms stimulate epithelial cells to produce inflammatory mediators through protease-activated receptor (PAR). We investigated the involvement of PAR2 in the interleukin (IL)-8 production by Helicobacter pylori-infected gastric epithelial cells. Methods and Results: Human gastric epithelial cells, MKN45 cells, were used. The expression of PAR2 was assessed by real-time PCR

Hirokazu Kajikawa; Norimasa Yoshida; Kazuhiro Katada; Fumihiro Hirayama; Osamu Handa; Satoshi Kokura; Yuji Naito; Toshikazu Yoshikawa

2007-01-01

356

Comparative study of the dynamics of focal contacts in live epithelial and mesenchymal cells  

Microsoft Academic Search

To further characterize the morphology and dynamics of focal contacts (FCs) in epithelial cells, we compared the size, number,\\u000a localization, velocity, and turnover of FCs in epithelial and mesenchymal cell lines. Using immunocytochemistry, we found\\u000a there were no significant differences between mesenchymal and epithelial cells in number and appearance whereas the location\\u000a and size of FCs in each cell were

Toshiyuki Ozawa; Daisuke Tsuruta

2011-01-01

357

How Bacteria-Induced Apoptosis of Intestinal Epithelial Cells Contributes to Mucosal Inflammation  

PubMed Central

The life cycle of an intestinal epithelial cell is terminated by apoptosis and/or cell shedding. Apoptotic deletion of epithelial cells from the intact intestinal mucosa is not accompanied by detectable inflammatory response or loss of barrier function. But increased permeability of the epithelial barrier and increased apoptotic rates of epithelial cells have been reported for patients suffering from inflammatory bowel disease. Microbiota can both induce or inhibit apoptosis of intestinal epithelial cells thus contribute to mucosal inflammation or support epithelial integrity respectively. Bacteria-mediated cytokine secretion and altered cell signalling are central to epithelial injury. Tumor necrosis factor (TNF) secreted after exposure to invasive bacteria induces both apoptosis and cell shedding. TNF is the major target gene of the transcription factor nuclear factor-kappa B with both pro- and anti-apoptotic effects. Autophagy promotes both cell survival and “autophagic” cell death. If autophagy is directed against microbes it is termed xenophagy. Inhibition of xenophagy has been shown to decrease cell survival. Endoplasmic reticulum (ER) stress causes misfolded proteins to accumulate in the ER lumen. It was suggested that ER stress and autophagy may interact within intestinal epithelial cells. Apoptosis in response to infection may be well proposed by the host to delete infected epithelial cells or could be a strategy of microbial pathogens to escape from exhausted cells to invade deeper mucosal layers for a prolonged bacterial colonization.

Hausmann, Martin

2010-01-01

358

Ex vivo culture of primary human fallopian tube epithelial cells.  

PubMed

Epithelial ovarian cancer is a leading cause of female cancer mortality in the United States. In contrast to other women-specific cancers, like breast and uterine carcinomas, where death rates have fallen in recent years, ovarian cancer cure rates have remained relatively unchanged over the past two decades (1). This is largely due to the lack of appropriate screening tools for detection of early stage disease where surgery and chemotherapy are most effective (2, 3). As a result, most patients present with advanced stage disease and diffuse abdominal involvement. This is further complicated by the fact that ovarian cancer is a heterogeneous disease with multiple histologic subtypes (4, 5). Serous ovarian carcinoma (SOC) is the most common and aggressive subtype and the form most often associated with mutations in the BRCA genes. Current experimental models in this field involve the use of cancer cell lines and mouse models to better understand the initiating genetic events and pathogenesis of disease (6, 7). Recently, the fallopian tube has emerged as a novel site for the origin of SOC, with the fallopian tube (FT) secretory epithelial cell (FTSEC) as the proposed cell of origin (8, 9). There are currently no cell lines or culture systems available to study the FT epithelium or the FTSEC. Here we describe a novel ex vivo culture system where primary human FT epithelial cells are cultured in a manner that preserves their architecture, polarity, immunophenotype, and response to physiologic and genotoxic stressors. This ex vivo model provides a useful tool for the study of SOC, allowing a better understanding of how tumors can arise from this tissue, and the mechanisms involved in tumor initiation and progression. PMID:21610668

Fotheringham, Susan; Levanon, Keren; Drapkin, Ronny

2011-05-09

359

Effects of. alpha. -particle radiation on rat tracheal epithelial cells  

SciTech Connect

By a combination of methods, which included flow cytometry and magnetic cell sorting, we have demonstrated that the cells of the rat tracheal epithelium which have the greatest proliferative capacity in culture and in vivo are the basal cells. Because of these findings it seems reasonable to suppose that the basal cells are the most likely target for the action of {alpha}-particle radiation in pseudostratified respiratory epithelium. This hypothesis is further supported by the finding that the basal cells are the cells which appear to respond to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. The effects of {sup 210}Po {alpha}-particles on the survival and oncogenic transformation of rat tracheal epithelial cells in suspension were investigated. Since these effects were assayed in culture, the results pertain to the reaction of only the basal cells to irradiation. The results indicate that {alpha}-particles are extremely cytotoxic in that a track segment of 4 {mu}m, on average, is sufficient to cause the reproductive death of basal cells. This finding is supported by similar results obtained with two cell lines, Mv1Lu and CHO-K1 BH{sub 4}. Production of proliferating epithelial foci by {alpha}-particles was not distinguishable from control and sham treatments. These results are in direct conflict with many of the results that have been obtained with C3H 1OT1/2 cells in similar transformation assays. Some possible reasons for these disparities are discussed and supporting evidence is provided.

Ford, J.R. Jr.

1992-08-01

360

Human epithelial cell cultures from superficial limbal explants  

PubMed Central

Purpose To study the kinetics of growth and the phenotype of cells cultured from human limbal explants in a cholera toxin-free medium with no feeder cell layer. Methods Human organ-cultured corneas were used to prepare limbal explants (full-thickness and superficial limbal explants) and corneal stromal explants. Cell growth kinetics and phenotypes were assessed by cultivating explants in cholera toxin-free Green medium. Epithelial and progenitor cell markers were assessed by immunocytochemistry, flow cytometry, and Reverse Transcription and Polymerase Chain Reaction (RT-PCR). Results The successful epithelial cell growth rates from full thickness limbal explant and superficial limbal explant tissues were 41 and 86%, respectively (p=0.0001). The mean cell area and the percentage of small cells in superficial and full-thickness explant cultures were, respectively, 317 µm2 and 429 µm2, and 8.9% and 1.7% (p<0.001). The percentage of positive cells in superficial and full-thickness limbal explant cultures as assessed by immunocytochemistry were the following: broad spectrum cytokeratins (cytokeratins 4, 5, 6, 8, 10, 13, and 18 [MNF116]), 82%/37% (p=0.01); cytokeratin 3 (CK3), 74%/25% (p=0.009); cytokeratin 19 (CK19), 46%/25% (p=0.19); vimentin, 56%/53% (p=0.48); delta N p63?, 54%/0% (p<0.001); and ABCG2, 5%/0% (p=0.1). Flow cytometry showed a higher percentage of small cells, a higher percentage of MNF116+ cells, and stronger expression of progenitor-associated markers in superficial than in full-thickness explant cultures. For superficial limbal explant cultures, analysis of the expression profiles for various mRNAs at the end of 21 days of culture showed high levels of expression of the mRNAs encoding CK3, vimentin, and CK19. The expression of mRNA of delta N p63? and ABCG2 was weaker. Cultures obtained from full-thickness limbal explants featured no expression of mRNA of CK19, delta N p63?, and ABCG2, whereas mRNAs encoding CK3 and vimentin were detected. Human corneal stromal explants cultured with the same medium featured late cell growth, large mean cell area (2,529 µm2), no expression of cytokeratins, delta N p63?, and ABCG2, and high expression of vimentin. Conclusions Superficial limbal explants appear to be superior to full-thickness limbal explants for growing human limbal epithelial cells. Preparation of explants using surgical facilities (i.e., operating microscope and microsurgical blades) led to a dramatic increase in the percentage of successful cultures, higher epithelial cell growth, decreased fibroblast contamination, and better preservation of limbal epithelial progenitors.

Basli, E.; Goldschmidt, P.; Pecha, F.; Chaumeil, C.; Laroche, L.; Borderie, V.

2011-01-01

361

Lung epithelial cells release ATP during ozone exposure: Signaling for cell survival  

Microsoft Academic Search

The common air pollutant ozone causes acute toxicity to human airways. In primary and transformed epithelial cells from all levels of human or rat airways, ozone levels relevant to air pollution (50–200 ppb) increased extracellular [ATP] within 7–30 min. A human bronchial epithelial cell line (16HBE14o-) that forms electrically resistant polarized monolayers had up to 10-fold greater apical than basolateral

Shama Ahmad; Aftab Ahmad; Glen McConville; Barbara K. Schneider; Corrie B. Allen; Rizwan Manzer; Robert J. Mason; Carl W. White

2005-01-01

362

Cell adhesion-related gene expression by Helicobacter pylori in gastric epithelial AGS cells  

Microsoft Academic Search

Helicobacter pylori (H. pylori) infection leads to gastroduodenal inflammation, peptic ulceration and gastric carcinoma. H. pylori may induce disease-specific gene expression in gastric epithelial cells. cDNA microarray for 352 cancer-related genes was used to identify the genes altered by H. pylori (cagA positive) in gastric epithelial AGS cells. Expressions of the genes identified on the microarray and other genes closely

Joo Weon Lim; Hyeyoung Kim; Kyung Hwan Kim

2003-01-01

363

UV-induced changes in cell cycle and gene expression within rabbit lens epithelial cells  

SciTech Connect

Damage to lens epithelial cells is a probable initiation process in cataract formation induced by ultraviolet radiation. These experiments investigated the ability of 254 nm radiation on cell cycle progression and gene expression in rabbit lens epithelial cell line N/N1003A. No changes in expression of c-fos, c-jun, alpha- tubulin, or vimentin was observed following UV exposure. Using flow cytometry, an accumulation of cells in G1/S phase of the cell cycle 1 hr following exposure. The observed changes in gene expression, especially the decreased histone transcripts reported here may play a role in UV induced inhibition of cell cycle progression.

Sidjanin, D. [Northern Illinois Univ., De Kalb, IL (United States). Dept. of Biological Sciences; Grdina, D. [Argonne National Lab., IL (United States); Woloschak, G.E. [Northern Illinois Univ., De Kalb, IL (United States). Dept. of Biological Sciences

1994-11-01

364

Bombesin-like peptide receptors in human bronchial epithelial cells.  

PubMed

Northern blot and RNAse protection assays previously failed to detect bombesin-like peptide (BLP) receptors in normal human lung tissue, but by RT/PCR cultured human bronchial epithelial (HBE) cells expressed all three BLP receptor subtypes, predominantly neuromedin B (NMB) receptor. By RT/PCR, we found expression of all three BLP receptor subtypes by human lung tissue and confirmed NMB receptor expression in six out of six HBE samples. However, transformed HBE BEAS B2B cells expressed only gastrin-releasing peptide (GRP) receptors; saturable, high-affinity (Kd = 3.5 nM) specific [125I]GRP binding confirmed functional GRP receptor, with M(r) = 75 kDa and immunologic cross-reactivity with GRP receptor from human small-cell lung carcinoma (SCLC) NCI-H345 cells. Altered regulation of BLP receptors may accompany transformation of normal lung cells to cancer. PMID:8822519

Kane, M A; Toi-Scott, M; Johnson, G L; Kelley, K K; Boose, D; Escobedo-Morse, A

1996-01-01

365

Differentiation of liver epithelial (stem-like) cells into hepatocytes induced by coculture with hepatic stellate cells  

Microsoft Academic Search

The liver is believed to contain stem cells that can differentiate into either hepatocytes or biliary epithelial cells. In the present study, we established a nonhepatocytic epithelial cell line from the normal livers of adult rats. The established cells, designated HSL cells, were immunoreactive against ?-fetoprotein, but neither albumin nor cytokeratin 19. To demonstrate the differentiation potential of HSL cells

Hirokazu Nagai; Kunihiko Terada; Go Watanabe; Yasuharu Ueno; Namiko Aiba; Tomomi Shibuya; Masami Kawagoe; Takashi Kameda; Mitsuru Sato; Haruki Senoo; Toshihiro Sugiyama

2002-01-01

366

MicroRNAs determine human intestinal epithelial cell fate  

PubMed Central

MicroRNAs (miRNAs) are small, non-coding RNA molecules that post-transcriptionally regulate gene expression. Evidence has shown that miRNAs play important roles in various cellular processes, including proliferation, differentiation and survival. The intestinal epithelium is regenerated throughout life, and enterocytes undergo differentiation during migration along the crypt/villus axis. Our study aimed at establishing the expression profiles of miRNAs during intestinal epithelial cell (IEC) differentiation and determining a miRNA “signature” that distinguishes between small and large IECs. MiRNA arrays were employed to profile miRNA expression in two IEC models: the enterocyte-like Caco2-BBE and the colonocyte-like HT29-Cl.19A cell lines. Microarray data showed that in both cell lineages, the differentiated stage exhibited a different miRNA expression profile from undifferentiated stage. Interestingly, Caco2-BBE cells were distinguished from HT29-Cl.19A cells by their unique miRNA expression profile. Notably, HT29-Cl.19A cells exhibited down-regulation of miR-1269 and up-regulation of miR-99b and miR-125a-5p compared with Caco2-BBE cells. Most importantly, transfection of Caco2-BBE cells with mature miR-99b, mature miR-125a-5p and antisense of mature miR-1269 decreased growth rate and trans-epithelial resistance of the cells, indicating their shift toward HT29-Cl.19A cell phenotype. In conclusion, our study shows that miRNAs might play a role in determining the unique physiological characteristics of IECs.

Dalmasso, Guillaume; Nguyen, Hang Thi Thu; Yan, Yutao; Laroui, Hamed; Srinivasan, Shanthi; Sitaraman, Shanthi V; Merlin, Didier

2010-01-01

367

Group B Streptococcus CovR regulation modulates host immune signalling pathways to promote vaginal colonization.  

PubMed

Streptococcus agalactiae (Group B?Streptococcus, GBS) is a frequent commensal organism of the vaginal tract of healthy women. However, GBS can transition to a pathogen in susceptible hosts, but host and microbial factors that contribute to this conversion are not well understood. GBS CovR/S (CsrR/S) is a two component regulatory system that regulates key virulence elements including adherence and toxin production. We performed global transcription profiling of human vaginal epithelial cells exposed to WT, CovR deficient, and toxin deficient strains, and observed that insufficient regulation by CovR and subsequent increased toxin production results in a drastic increase in host inflammatory responses, particularly in cytokine signalling pathways promoted by IL-8 and CXCL2. Additionally, we observed that CovR regulation impacts epithelial cell attachment and intracellular invasion. In our mouse model of GBS vaginal colonization, we further demonstrated that CovR regulation promotes vaginal persistence, as infection with a CovR deficient strainresulted in a heightened host immune response as measured by cytokine production and neutrophil activation. Using CXCr2 KO mice, we determined that this immune alteration occurs, at least in part, via signalling through the CXCL2 receptor. Taken together, we conclude that CovR is an important regulator of GBS vaginal colonization and loss of this regulatory function may contribute to the inflammatory havoc seen during the course of infection. PMID:23298320

Patras, Kathryn A; Wang, Nai-Yu; Fletcher, Erin M; Cavaco, Courtney K; Jimenez, Alyssa; Garg, Mansi; Fierer, Joshua; Sheen, Tamsin R; Rajagopal, Lakshmi; Doran, Kelly S

2013-01-30

368

Origin and phenotypic features of hyperplastic epithelial cells in collapsing glomerulopathy  

Microsoft Academic Search

The characteristic pathological feature of collapsing glomerulopathy (CG) is marked cell hyperplasia and hypertrophy within the glomeruli. The present study investigated the phenotypic alteration of hyperplastic epithelial cells in CG to determine their origin. Renal biopsy specimens from two patients with CG were analyzed by immunohistochemical staining, using markers for podocytes (PHM-5), parietal epithelial cells (PECs; cytokeratin), and cell proliferation

M Nagata; M Hattori; Y Hamano; K Ito; K Saitoh; T Watanabe

1998-01-01

369

Secretion of cytokines and chemokines by polarized human epithelial cells from the female reproductive tract  

Microsoft Academic Search

BACKGROUND: Pro-inflammatory chemokines that attract and cytokines that activate immune cells contribute to normal physiological homeostasis in the female reproductive tract, and are needed to deal effectively with poten- tial pathogenic microbes. Mucosal epithelial cells are capable of producing these factors that communicate with cells of the innate and adaptive immune systems. METHODS: Epithelial cells from Fallopian tube, endometrium and

J. V. Fahey; T. M Schaefer; J. Y. Channon; C. R. Wira

2005-01-01

370

A Common Epithelial Cell Surface Antigen (EPM-1) on Gastrointestinal Tumors and in Human Sera1  

Microsoft Academic Search

A commonepithelial cell surface marker (EPM-1) was defined by two monoclonal antibodies (Pa-25 and Pa-42), raised against a pancreatic tumor cell line (Capan-1). Both monoclonal antibodies were tested on 49 cultured human cell lines and 244 tissue samples and reacted with all 76 tissue samples of 20 different normal epithelial cell types and with 57 of 63 epithelial tumor tissue

Wolfgang G. Dippold; Helga Bernhard; Reinhard Klingel; Hans-Peter Dienes; Birgit Schneider; Alexander Knuth; Karl-Hermann Meyer

371

Activated endothelial cells elicit paracrine induction of epithelial chloride secretion. 6-Keto-PGF1alpha is an epithelial secretagogue.  

PubMed Central

Endothelial cells play a central role in the coordination of the inflammatory response. In mucosal tissue, such as the lung and intestine, endothelia are anatomically positioned in close proximity to epithelia, providing the potential for cell-cell crosstalk. Thus, in this study endothelial-epithelial biochemical crosstalk pathways were studied using a human intestinal crypt cell line (T84) grown in noncontact coculture with human umbilical vein endothelia. Exposure of such cocultures to endothelial-specific agonists (LPS) resulted in activation of epithelial electrogenic Cl- secretion and vectorial fluid transport. Subsequent experiments revealed that in response to diverse stimuli (LPS, IL-1alpha, TNF-alpha, hypoxia), endothelia produce and secrete a small, stable epithelial secretagogue into conditioned media supernatants. Further experiments identified this secretagogue as 6-keto-PGF1alpha, a stable hydrolysis product of prostacyclin (PGI2). Results obtained with synthetic prostanoids indicated that 6-keto-PGF1alpha (EC50 = 80 nM) and PGI2 stable analogues (EC50 = 280 nM) activate the same basolaterally polarized, Ca2+-coupled epithelial receptor. In summary, these findings reveal a previously unappreciated 6-keto-PGF1alpha receptor on intestinal epithelia, the ligation of which results in activation of electrogenic Cl- secretion. In addition, these data reveal a novel action for the prostacyclin hydrolysis product 6-keto-PGF1alpha and provide a potential endothelial- epithelial crosstalk pathway in mucosal tissue.

Blume, E D; Taylor, C T; Lennon, P F; Stahl, G L; Colgan, S P

1998-01-01

372

Hypoxia Modulates Infection of Epithelial Cells by Pseudomonas aeruginosa  

PubMed Central

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen commonly associated with lung and wound infections. Hypoxia is a frequent feature of the microenvironment of infected tissues which induces the expression of genes associated with innate immunity and inflammation in host cells primarily through the activation of the hypoxia-inducible factor (HIF) and Nuclear factor kappaB (NF-?B) pathways which are regulated by oxygen-dependent prolyl-hydroxylases. Hypoxia also affects virulence and antibiotic resistance in bacterial pathogens. However, less is known about the impact of hypoxia on host-pathogen interactions such as bacterial adhesion and infection. In the current study, we demonstrate that hypoxia decreases the internalization of P. aeruginosa into cultured epithelial cells resulting in decreased host cell death. This response can also be elicited by the hydroxylase inhibitor Dimethyloxallyl Glycine (DMOG). Reducing HIF-2? expression or Rho kinase activity diminished the effects of hypoxia on P. aeruginosa infection. Furthermore, in an in vivo pneumonia infection model, application of DMOG 48 h before infection with P. aeruginosa significantly reduced mortality. Thus, hypoxia reduces P. aeruginosa internalization into epithelial cells and pharmacologic manipulation of the host pathways involved may represent new therapeutic targets in the treatment of P. aeruginosa infection.

Schaible, Bettina; McClean, Siobhan; Selfridge, Andrew; Broquet, Alexis; Asehnoune, Karim

2013-01-01

373

OCRL1 Modulates Cilia Length in Renal Epithelial Cells  

PubMed Central

Lowe syndrome is an X-linked disorder characterized by cataracts at birth, mental retardation and progressive renal malfunction that results from loss of function of the OCRL1 (oculocerebrorenal syndrome of Lowe) protein. OCRL1 is a lipid phosphatase that converts phosphatidylinositol 4,5-bisphosphate to phosphatidylinositol 4-phosphate. The renal pathogenesis of Lowe syndrome patients has been suggested to result from alterations in membrane trafficking, but this cannot fully explain the disease progression. We found that knockdown of OCRL1 in zebrafish caused developmental defects consistent with disruption of ciliary function, including body axis curvature, pericardial edema, hydrocephaly and impaired renal clearance. In addition, cilia in the proximal tubule of the zebrafish pronephric kidney were longer in ocrl morphant embryos. We also found that knockdown of OCRL1 in polarized renal epithelial cells caused elongation of the primary cilium and disrupted formation of cysts in three-dimensional cultures. Calcium release in response to ATP was blunted in OCRL1 knockdown cells, suggesting changes in signaling that could lead to altered cell function. Our results suggest a new role for OCRL1 in renal epithelial cell function that could contribute to the pathogenesis of Lowe syndrome.

Rbaibi, Youssef; Cui, Shanshan; Mo, Di; Carattino, Marcelo; Rohatgi, Rajeev; Satlin, Lisa M.; Szalinski, Christina M.; Swanhart, Lisa M.; Folsch, Heike; Hukriede, Neil A.; Weisz, Ora A.

2013-01-01

374

Aquaporin 2 Promotes Cell Migration and Epithelial Morphogenesis  

PubMed Central

The aquaporin 2 (AQP2) water channel, expressed in kidney collecting ducts, contributes critically to water homeostasis in mammals. Animals lacking or having significantly reduced levels of AQP2, however, have not only urinary concentrating abnormalities but also renal tubular defects that lead to neonatal mortality from renal failure. Here, we show that AQP2 is not only a water channel but also an integrin?binding membrane protein that promotes cell migration and epithelial morphogenesis. AQP2 expression modulates the trafficking and internalization of integrin ?1, facilitating its turnover at focal adhesions. In vitro, disturbing the interaction between AQP2 and integrin ?1 by mutating the RGD motif led to reduced endocytosis, retention of integrin ?1 at the cell surface, and defective cell migration and tubulogenesis. Similarly, in vivo, AQP2?null mice exhibited significant retention of integrin ?1 at the basolateral membrane and had tubular abnormalities. In summary, these data suggest that the water channel AQP2 interacts with integrins to promote renal epithelial cell migration, contributing to the structural and functional integrity of the mammalian kidney.

Chen, Ying; Rice, William; Gu, Zhizhan; Li, Jian; Huang, Jianmin; Brenner, Michael B.; Van Hoek, Alfred; Xiong, Jianping; Gundersen, Gregg G.; Norman, Jim C.; Hsu, Victor W.; Fenton, Robert A.; Brown, Dennis

2012-01-01

375

Vaginal gel formulation based on theaflavin derivatives as a microbicide to prevent HIV sexual transmission.  

PubMed

We previously demonstrated that a commercially available natural product preparation with high content (>90%) of theaflavin derivatives (TFmix) exhibited potent anti-HIV activities. Here we developed a TFmix gel formulation as a topical microbicide candidate. The effect of TFmix on the amyloid fibril formation of semen-derived enhancer of virus infection (SEVI) peptide was detected by transmission electron microscopy. The toxicity of the TFmix gel was evaluated using human vaginal and cervical epithelial cell lines and rabbit vaginal irritation models, respectively. Levels of proinflammatory cytokines (IL-1?, IL-6, IL-8, and TNF-?) and immunoregulatory cytokines (IL-10 and GM-CSF) in cervicovaginal lavages (CVLs) were measured by ELISA kits. Proliferating cell nuclear antigen (PCNA) immunostaining was performed to evaluate inflammation in the vaginal tissues. TFmix gel could degrade SEVI-specific amyloid fibrils and showed low cytotoxicity to epithelial cells of the female reproductive tract. No apparent cervicovaginal toxicity was observed at any time point evaluated following the intravaginal administration of TFmix gel to rabbits, whereas application of N-9 gel resulted in damage to the vaginal epithelium. Neither proinflammatory nor immunoregulatory cytokine production was triggered by TFmix gel. Only low expression of PCNA was observed in vaginal tissues of TFmix gel-treated rabbits. The concentration of TFmix in plasma was very low (below the lower limit of quantitation) 1?h after a single vaginal administration of TFmix gel. However, TFmix was still detected in the cervicovaginal lavages (CVLs) 6?h after treatment, indicating that it could be retained in the vaginal cavity for a long period of time. With its potent anti-HIV-1 activity, marked stability at acidic condition, low mucosal toxicity, and lack of systemic absorption, TFmix gel can be considered as an inexpensive and safe microbicide candidate for the prevention of HIV sexual transmission. PMID:22867271

Yang, Jie; Li, Lin; Jin, Hong; Tan, Suiyi; Qiu, Jiayin; Yang, Lei; Ding, Yanqing; Jiang, Zhi-Hong; Jiang, Shibo; Liu, Shuwen

2012-09-11

376

Culture and characterization of oral mucosal epithelial cells on human amniotic membrane for ocular surface reconstruction  

PubMed Central

Purpose To culture oral mucosal epithelial cells on deepithelialized human amniotic membrane without the use of feeder cells and to compare the characteristics of cultured oral cells with cultured limbal and conjunctival epithelial cells for use in ocular surface reconstruction. Methods Oral biopsies were obtained from healthy volunteers after informed consent and were cultured on deepithelialized amniotic membrane for three to four weeks. Confluent cultures of limbal, oral, and conjunctival cells were subjected to characterization of markers of stem cells and of epithelial differentiation by reverse-transcription polymerase chain reaction (RT–PCR) and by immunohistochemistry. Ultrastructural studies were also performed using electron microscopy. Results A sheet of healthy, stratified oral epithelial cells was obtained within three to four weeks of culture. Electron microscopy demonstrated that the cells formed gap junctions and desmosomes. RT–PCR analysis showed that cultured oral epithelial cells expressed markers of epithelial differentiation such as cytokeratin K3, K4, K13, K15 and connexin 43. The cells also expressed stem cell markers of epithelial cells such as ?N isoforms of p63 as well as p75, a marker for stem cells of oral epithelium. The cells did not express cytokeratin K12 or Pax-6, an eye-specific transcription factor. Conclusions Oral epithelial cells can be cultured as explants on deepithelialized amniotic membrane without using feeder cells. Characterization showed that these cells maintain the phenotypic characteristics of oral epithelial cells and that the culture is a heterogeneous population of differentiated cells and stem cells. We find the cultured oral epithelial cells usable for ocular surface reconstruction in patients having bilateral ocular surface diseases.

Madhira, Soundarya Lakshmi; Vemuganti, Geeta; Bhaduri, Anirban; Gaddipati, Subhash; Sangwan, Virender Singh

2008-01-01

377

Survivin expression is associated with lens epithelial cell proliferation and fiber cell differentiation  

PubMed Central

Purpose Survivin (Birc5) is the smallest member of the inhibitor of apoptosis (IAP) protein family, which regulates the cell cycle/apoptosis balance. The purpose of this study was to examine Survivin expression in the embryonic chick lens, in chick lens epithelial cell cultures, and in the postnatal mouse lens. Methods Survivin expression was examined using a combination of quantitative real-time polymerase chain reaction, western blotting, and immunocytochemistry. To correlate Survivin expression with the timing of proliferation, we determined the profile of cell proliferation in the developing lens using the cell cycle marker proliferating cell nuclear antigen (PCNA) in quantitative western blotting and immunocytochemistry studies. We also examined the expression of PCNA and the extent of denucleation using terminal deoxynucleotidyl transferase (TdT)-mediated biotin-dUTP nick-end labeling (TUNEL) of lentoids (lens fiber-like cells) during chick lens epithelial cell differentiation in vitro. Results At embryonic day (ED) 4, Survivin immunostaining was present in two pools in lens epithelial cells and fiber cells: cytoplasmic and nuclear. The nuclear staining became more pronounced as the lens epithelial cells differentiated into lens fiber cells. At ED12, Survivin staining was observed in lens fiber cell nuclei containing marginalized chromatin, indicative of early denucleation events. Using western blotting, Survivin expression peaked at ED6, diminishing thereafter. This profile of expression correlated with the events in chick lens epithelial cell cultures: i) increased Survivin expression was associated with an increase in PCNA staining up to day 6 of culture and ii) downregulation of Survivin expression at day 8 of culture was coincident with a dramatic decrease in PCNA staining and an increase in TdT-mediated biotin-dUTP nick-end labeling in lentoids. In early postnatal mouse lenses, Survivin and PCNA were highly expressed and decreased thereafter during postnatal lens maturation. Conclusions Survivin is expressed during chick and mouse lens development and in chick lens epithelial cell cultures. High levels of Survivin expression correlated with high rates of proliferation of lens epithelial cells at early stages of development. Downregulation of Survivin expression with development and its progressive localization to the nuclei of lens fiber cells was coincident with a decrease in cell proliferation and increased denucleation in differentiating lens fiber cells. These studies suggest an important role for Survivin as a dual regulator of lens epithelial cell proliferation and lens fiber cell differentiation.

Mansergh, Fiona C.; Boulton, Michael E.; Gunhaga, Lena

2012-01-01

378

CARD15\\/NOD2 functions as an antibacterial factor in human intestinal epithelial cells  

Microsoft Academic Search

Background & Aims: Mutations in the CARD15\\/NOD2 gene, a putative intracellular pattern recognition receptor, have been linked to the risk for Crohn's disease. Because intestinal epithelial cells play a role as the barrier to luminal microorganisms, we investigated the expression and function of CARD15\\/NOD2 in intestinal epithelial cells. Methods: Expression of CARD15\\/NOD2 messenger RNA (mRNA) in intestinal epithelial cell lines

Tadakazu Hisamatsu; Manabu Suzuki; Hans-Christian Reinecker; William J. Nadeau; Beth A. McCormick; Daniel K. Podolsky

2003-01-01

379

Efficient Immortalization of Primary Nasopharyngeal Epithelial Cells for EBV Infection Study  

PubMed Central

Nasopharyngeal carcinoma (NPC) is common among southern Chinese including the ethnic Cantonese population living in Hong Kong. Epstein-Barr virus (EBV) infection is detected in all undifferentiated type of NPC in this endemic region. Establishment of stable and latent EBV infection in premalignant nasopharyngeal epithelial cells is an early event in NPC development and may contribute to its pathogenesis. Immortalized primary nasopharyngeal epithelial cells represent an important tool for investigation of EBV infection and its tumorigenic potential in this special type of epithelial cells. However, the limited availability and small sizes of nasopharyngeal biopsies have seriously restricted the establishment of primary nasopharyngeal epithelial cells for immortalization. A reliable and effective method to immortalize primary nasopharyngeal epithelial cells will provide unrestricted materials for EBV infection studies. An earlier study has reported that Bmi-1 expression could immortalize primary nasopharyngeal epithelial cells. However, its efficiency and actions in immortalization have not been fully characterized. Our studies showed that Bmi-1 expression alone has limited ability to immortalize primary nasopharyngeal epithelial cells and additional events are often required for its immortalization action. We have identified some of the key events associated with the immortalization of primary nasopharyngeal epithelial cells. Efficient immortalization of nasopharyngeal epithelial cells could be reproducibly and efficiently achieved by the combined actions of Bmi-1 expression, activation of telomerase and silencing of p16 gene. Activation of MAPK signaling and gene expression downstream of Bmi-1 were detected in the immortalized nasopharyngeal epithelial cells and may play a role in immortalization. Furthermore, these newly immortalized nasopharyngeal epithelial cells are susceptible to EBV infection and supported a type II latent EBV infection program characteristic of EBV-infected nasopharyngeal carcinoma. The establishment of an efficient method to immortalize primary nasopharyngeal epithelial cells will facilitate the investigation into the role of EBV infection in pathogenesis of nasopharyngeal carcinoma.

Yip, Yim Ling; Pang, Pei Shin; Deng, Wen; Tsang, Chi Man; Zeng, Musheng; Hau, Pok Man; Man, Cornelia; Jin, Yuesheng; Yuen, Anthony Po Wing; Tsao, Sai Wah

2013-01-01

380

Role of Porphyromonas gingivalis SerB in Gingival Epithelial Cell Cytoskeletal Remodeling and Cytokine Production  

Microsoft Academic Search

The SerB protein of Porphyromonas gingivalis is a HAD family serine phosphatase that plays a critical role in entry and survival of the organism in gingival epithelial cells. SerB is secreted by P. gingivalis upon contact with epithelial cells. Here it is shown by microarray analysis that SerB impacts the transcriptional profile of gingival epithelial cells, with pathways involving the

Yoshiaki Hasegawa; Gena D. Tribble; Henry V. Baker; Jeffrey J. Mans; Martin Handfield; Richard J. Lamont

2008-01-01

381

Sorting of synaptophysin into special vesicles in nonneuroendocrine epithelial cells  

PubMed Central

Synaptophysin is a major transmembrane glycoprotein of a type of small vesicle with an electron-translucent content (SET vesicles), including the approximately 50-nm presynaptic vesicles in neuronal cells, and of similar, somewhat larger (< or = approximately 90 nm) vesicles (SLMV) in neuroendocrine (NE) cells. When certain epithelial non-NE cells, such as human hepatocellular carcinoma PLC cells, were cDNA transfected to synthesize synaptophysin, the new molecules appeared in specific SET vesicles. As this was in contrast to other reports that only NE cells were able to sort synaptophysin away from other plasma membrane proteins into presynaptic- or SLMV-type vesicles, we have further characterized the vesicles containing synaptophysin in transfected PLC cells. Using fractionation and immunoisolation techniques, we have separated different kinds of vesicles, and we have identified a distinct type of synaptophysin-rich, small (30-90-nm) vesicle that contains little, if any, protein of the constitutive secretory pathway marker hepatitis B surface antigen, of the fluid phase endocytosis marker HRP, and of the plasma membrane recycling endosomal marker transferrin receptor. In addition, we have found variously sized vesicles that contained both synaptophysin and transferrin receptor. A corresponding result was also obtained by direct visualization, using double-label immunofluorescence microscopy for the endocytotic markers and synaptophysin in confocal laser scan microscopy and in double- immunogold label electron microscopy. We conclude that diverse non-NE cells of epithelial nature are able to enrich the "foreign" molecule synaptophysin in a category of SET vesicles that are morphologically indistinguishable from SLMV of NE cells, including one type of vesicle in which synaptophysin is sorted away from endosomal marker proteins. Possible mechanisms of this sorting are discussed.

1994-01-01

382

Phototoxicity and cytotoxicity of fullerol in human lens epithelial cells  

SciTech Connect

The water-soluble, hydroxylated fullerene [fullerol, nano-C{sub 60}(OH){sub 22-26}] has several clinical applications including use as a drug carrier to bypass the blood ocular barriers. We have assessed fullerol's potential ocular toxicity by measuring its cytotoxicity and phototoxicity induced by UVA and visible light in vitro with human lens epithelial cells (HLE B-3). Accumulation of nano-C{sub 60}(OH){sub 22-26} in the cells was confirmed spectrophotometrically at 405 nm and cell viability estimated using MTS and LDH assays. Fullerol was cytotoxic to HLE B-3 cells maintained in the dark at concentrations higher than 20 {mu}M. Exposure to either UVA or visible light in the presence of > 5 {mu}M fullerol-induced phototoxic damage. When cells were pretreated with non-toxic antioxidants: 20 {mu}M lutein, 1 mM N-acetyl cysteine, or 1 mM L-ascorbic acid prior to irradiation, only the singlet oxygen quencher-lutein significantly protected against fullerol photodamage. Apoptosis was observed in lens cells treated with fullerol whether or not the cells were irradiated, in the order UVA > visible light > dark. Dynamic light scattering (DLS) showed that in the presence of the endogenous lens protein {alpha}-crystallin, large aggregates of fullerol were reduced. In conclusion, fullerol is both cytotoxic and phototoxic to human lens epithelial cells. Although the acute toxicity of water-soluble nano-C{sub 60}(OH){sub 22-26} is low, these compounds are retained in the body for long periods, raising concern for their chronic toxic effect. Before fullerols are used to deliver drugs to the eye, they should be tested for photo- and cytotoxicity in vivo.

Roberts, Joan E. [Department of Natural Sciences, Fordham University, 113 West 60th Street, New York City, NY 10023 (United States)], E-mail: jroberts@fordham.edu; Wielgus, Albert R. [Laboratory of Pharmacology and Chemistry, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709 (United States)], E-mail: wielgus@niehs.nih.gov; Boyes, William K. [Neurotoxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711 (United States)], E-mail: Boyes.William@epamail.epa.gov; Andley, Usha [Department of Ophthalmology and Visual Science, Washington University School of Medicine, St. Louis, MO 63110 (United States)], E-mail: andley@vision.wustl.edu; Chignell, Colin F. [Laboratory of Pharmacology and Chemistry, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709 (United States)], E-mail: chignell@niehs.nih.gov

2008-04-01

383

Epithelial-mesenchymal transition, cancer stem cells and treatment resistance  

PubMed Central

Breast cancer relapse, in a large number of patients, after initial response to standard of care therapy warrants development of novel therapies against recurrent and metastatic cancer. Cancer stem cells (CSCs), present in breast tumors while being intrinsically resistant to conventional therapy, have the ability to self renew and cause tumor recurrence. The residual tumors after therapy, with dramatic enrichment of the CSCs, have all the hallmarks of epithelial- mesenchymal transition (EMT). This review will focus on the link between EMT, CSCs and treatment resistance, since a better understanding of these interactions will allow us to effectively target the residual population after therapy.

2012-01-01