scholarly journals Bovine herpesvirus 4 is tropic for bovine endometrial cells and modulates endocrine function

Reproduction ◽  
2007 ◽  
Vol 134 (1) ◽  
pp. 183-197 ◽  
Author(s):  
Gaetano Donofrio ◽  
Shan Herath ◽  
Chiara Sartori ◽  
Sandro Cavirani ◽  
Cesidio Filippo Flammini ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Bovine Herpesvirus ◽  
Endocrine Function ◽  
Uterine Disease ◽  
Endometrial Stromal Cells ◽  
Persistently Infected ◽  
Endometrial Cells ◽  
Bovine Herpesvirus 4 ◽  
Herpesvirus 4

Bovinepostpartumuterine disease, metritis, affects about 40% of animals and is widely considered to have a bacterial aetiology. Although the γ-herpesvirus bovine herpesvirus 4 (BoHV-4) has been isolated from several outbreaks of metritis or abortion, the role of viruses in endometrial pathology and the mechanisms of viral infection of uterine cells are often ignored. The objectives of the present study were to explore the interaction, tropism and outcomes of BoHV-4 challenge of endometrial stromal and epithelial cells. Endometrial stromal and epithelial cells were purified and infected with a recombinant BoHV-4 carrying an enhanced green fluorescent protein (EGFP) expression cassette to monitor the establishment of infection. BoHV-4 efficiently infected both stromal and epithelial cells, causing a strong non-apoptotic cytopathic effect, associated with robust viral replication. The crucial step for the BoHV-4 endometriotropism appeared to be after viral entry as there was enhanced transactivation of the BoHV-4 immediate early 2 gene promoter following transient transfection into the endometrial cells. Infection with BoHV-4 increased cyclooxygenase 2 protein expression and prostaglandin estradiol secretion in endometrial stromal cells, but not epithelial cells. Bovine macrophages are persistently infected with BoHV-4, and co-culture with endometrial stromal cells reactivated BoHV-4 replication in the persistently infected macrophages, suggesting a symbiotic relationship between the cells and virus. In conclusion, the present study provides evidence of cellular and molecular mechanisms, supporting the concept that BoHV-4 is a pathogen associated with uterine disease.

scholarly journals Bacterial infection of endometrial stromal cells influences bovine herpesvirus 4 immediate early gene activation: a new insight into bacterial and viral interaction for uterine disease

Reproduction ◽  
2008 ◽  
Vol 136 (3) ◽  
pp. 361-366 ◽  
Author(s):  
Gaetano Donofrio ◽  
Lara Ravanetti ◽  
Sandro Cavirani ◽  
Shan Herath ◽  
Antonio Capocefalo ◽  
...  
Keyword(s):  
Gene Activation ◽  
Bovine Herpesvirus ◽  
Early Gene ◽  
Immediate Early ◽  
Uterine Disease ◽  
Prostaglandin E ◽  
Endometrial Stromal Cells ◽  
Endometrial Cells ◽  
Bovine Herpesvirus 4 ◽  
Herpesvirus 4

Experimental infection with the γ-herpesvirus bovine herpesvirus 4 (BoHV-4) rarely establishes disease, yet BoHV-4 is commonly associated with uterine disease in cattle. Uterine disease involves co-infection with bacteria such asEscherichia coli, which stimulate the production of prostaglandin E2(PGE2) by endometrial cells. BoHV-4 replication depends on immediate early 2 (IE2) gene transactivation and, in the present study, PGE2,E. colior its lipopolysaccharide upregulated theIE2gene promoter in uterine cells. Bacterial co-infection is important for BoHV-4 uterine disease.

scholarly journals Endometrial Stromal Cells Circulate in the Bloodstream of Women with Endometriosis: A Pilot Study

2019 ◽  
Vol 20 (15) ◽  
pp. 3740 ◽  
Author(s):  
Júlia Vallvé-Juanico ◽  
Carlos López-Gil ◽  
Agustín Ballesteros ◽  
Xavier Santamaria
Keyword(s):  
Pilot Study ◽  
Epithelial Cells ◽  
Stromal Cells ◽  
Diagnostic Tools ◽  
Pelvic Cavity ◽  
Endometrial Stromal Cells ◽  
Endometrial Cells ◽  
Retrograde Menstruation ◽  
The Brain ◽  
Endometrial Epithelial Cells

Endometriosis is characterized by the presence of endometrial tissue outside the uterus. While endometriotic tissue is commonly localized in the pelvic cavity, it can also be found in distant sites, including the brain. The origin and pathophysiology of tissue migration is poorly understood; retrograde menstruation is thought to be the cause, although the presence of endometrium at distant sites is not explained by this hypothesis. To determine whether dissemination occurs via the bloodstream in women with endometriosis, we analyzed circulating blood for the presence of endometrial cells. Circulating endometrial stromal cells were identified only in women with endometriosis but not in controls, while endometrial epithelial cells were not identified in the circulation of either group. Our results support the hypothesis that endometrial stromal cells may migrate through circulation and promote the pathophysiology of endometriosis. The detection of these cells in circulation creates avenues for the development of less invasive diagnostic tools for the disease, and opens possibilities for further study of the origin of endometriosis.

scholarly journals Expression and Function of Toll-Like Receptor 4 in the Endometrial Cells of the Uterus

Endocrinology ◽  
2006 ◽  
Vol 147 (1) ◽  
pp. 562-570 ◽  
Author(s):  
Shan Herath ◽  
Deborah P. Fischer ◽  
Dirk Werling ◽  
Erin J. Williams ◽  
Sonia T. Lilly ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Prostaglandin F2α ◽  
Endocrine Function ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Endometrial Cells ◽  
E Coli ◽  
Female Sex Hormones ◽  
And Function

Prostaglandins have a central role in many endocrine functions in mammals, including regulation of the life span of the corpus luteum by prostaglandin F2α (PGF) and prostaglandin E2 (PGE), which are secreted by the uterine endometrium. However, the uterus is readily infected with bacteria such as Escherichia coli, which disrupt luteolysis. Immune cells detect E. coli by Toll-like receptor 4 (TLR4) binding its pathogenic ligand, lipopolysaccharide (LPS), although signaling requires accessory molecules such as CD14. The objective of this study was to determine the effect of E. coli or LPS on the function of bovine endometrial cells, and whether purified populations of epithelial and stromal cells express the molecules involved in LPS recognition. In addition, because the female sex hormones estradiol and progesterone modify the risk of uterine infection, their effect on the LPS response was investigated. Endometrial explants produced prostaglandins in response to LPS, with an increased ratio of PGE to PGF. Addition of LPS or E. coli to stromal and epithelial cells stimulated production of PGE and PGF and increased their cyclooxygenase 2 mRNA expression. The production of prostaglandins was abrogated by an LPS antagonist. In addition, estradiol and progesterone inhibited the production of PGE and PGF in response to LPS, indicating a role for steroid hormones in the response to bacterial infection. For the first time, Toll-like receptor 4 mRNA and CD14 mRNA and protein were detected in bovine endometrial stromal and epithelial cells by RT-PCR and flow cytometry. In conclusion, epithelial and stromal cells detect and respond to bacteria, which modulate their endocrine function.

scholarly journals Bacterial Lipopolysaccharide Induces an Endocrine Switch from Prostaglandin F2α to Prostaglandin E2 in Bovine Endometrium

Endocrinology ◽  
2008 ◽  
Vol 150 (4) ◽  
pp. 1912-1920 ◽  
Author(s):  
Shan Herath ◽  
Sonia T. Lilly ◽  
Deborah P. Fischer ◽  
Erin J. Williams ◽  
Hilary Dobson ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Prostaglandin E2 ◽  
Stromal Cells ◽  
Inflammatory Responses ◽  
Prostaglandin F2α ◽  
Uterine Disease ◽  
Group Vi ◽  
Endometrial Cells ◽  
Escherichia Coli Infection ◽  
Arachadonic Acid

Escherichia coli infection of the endometrium causes uterine disease after parturition and is associated with prolonged luteal phases of the ovarian cycle in cattle. Termination of the luteal phase is initiated by prostaglandin F2α (PGF) from oxytocin-stimulated endometrial epithelial cells. Compared with normal animals, the peripheral plasma of animals with E. coli infection of the endometrium had higher concentrations of lipopolysaccharide (LPS) and prostaglandin E2 (PGE) but not PGF. Endometrial explants accumulated predominantly PGE in the culture medium in response to LPS, and this effect was not reversed by oxytocin. Endometrial cells expressed the Toll-like receptor 4/CD14/MD-2 receptor complex necessary to detect LPS. Epithelial and stromal cells treated with LPS had higher steady-state media concentrations of PGE rather than PGF. Arachadonic acid is liberated from cell membranes by phospholipase 2 (PLA2) enzymes and converted to prostaglandins by synthase enzymes. Treatment of epithelial and stromal cells with LPS did not change the levels of PGE or PGF synthase enzymes. However, LPS stimulated increased levels of PLA2 group VI but not PLA2 group IV C immunoreactive protein in epithelial cells. Endometrial cells expressed the E prostanoid 2 and E prostanoid 4 receptors necessary to respond to PGE, which regulates inflammation as well as being luteotropic. In conclusion, LPS detection by endometrial cells stimulated the accumulation of PGE rather than PGF, providing a mechanism to explain prolonged luteal phases in animals with uterine disease, and this PGE may also be important for regulating inflammatory responses in the endometrium.

Swine adipose stromal cells loaded with recombinant bovine herpesvirus 4 virions expressing a foreign antigen induce potent humoral immune responses in pigs

Vaccine ◽  
2011 ◽  
Vol 29 (5) ◽  
pp. 867-872 ◽  
Author(s):  
Gaetano Donofrio ◽  
Simone Taddei ◽  
Valentina Franceschi ◽  
Antonio Capocefalo ◽  
Sandro Cavirani ◽  
...  
Keyword(s):  
Immune Responses ◽  
Stromal Cells ◽  
Bovine Herpesvirus ◽  
Adipose Stromal Cells ◽  
Foreign Antigen ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Bovine Herpesvirus 4 ◽  
Herpesvirus 4

scholarly journals A bovine macrophage cell line supports bovine herpesvirus-4 persistent infection

2001 ◽  
Vol 82 (5) ◽  
pp. 1181-1185 ◽  
Author(s):  
Gaetano Donofrio ◽  
Vicky L. van Santen
Keyword(s):  
Viral Genome ◽  
Bovine Herpesvirus ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Persistently Infected ◽  
Bovine Herpesvirus 4 ◽  
Bovine Macrophage ◽  
Herpesvirus 4

Although bovine herpesvirus-4 (BHV-4), a gammaherpesvirus lacking a clear disease association, has been demonstrated in many tissues during persistent BHV-4 infection, a likely site of virus persistence is in cells of the monocyte/macrophage lineage. To establish an in vitro model of persistent infection potentially useful for examining the molecular mechanisms of BHV-4 persistence/latency, we infected the bovine macrophage cell line BOMAC. Following extensive cell death, surviving cells were found to be persistently infected, maintaining the viral genome over many passages and producing low levels of infectious virus. Although selection was unnecessary for the maintenance of the viral genome, cells persistently infected with recombinant BHV-4 containing a neomycin-resistance gene could be selected with geneticin, thus confirming that persistent BHV-4 infection was compatible with cell survival and replication. Furthermore, persistent BHV-4 infection caused no decrease in the growth rate of BOMAC cells. Sodium butyrate, which reactivates latent gammaherpesviruses in vitro, or dexamethasone, which reactivates latent BHV-4 in vivo, increased viral DNA by 10- to 15-fold in persistently infected BOMAC cells. This suggests that reactivation of latent BHV-4 by dexamethasone in vivo might involve direct action of dexamethasone on latently infected cells.

Establishment of a cell line persistently infected with bovine herpesvirus-4 by use of a recombinant virus

Microbiology ◽  
2000 ◽  
Vol 81 (7) ◽  
pp. 1807-1814 ◽  
Author(s):  
Gaetano Donofrio ◽  
Sandro Cavirani ◽  
Vicky L. van Santen
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Viral Genome ◽  
Infectious Virus ◽  
Viral Gene Expression ◽  
Bovine Herpesvirus ◽  
Viral Gene ◽  
Persistently Infected ◽  
Bovine Herpesvirus 4 ◽  
Herpesvirus 4

Bovine herpesvirus-4 (BHV-4), a gammaherpesvirus lacking a clear disease association, productively infects multiple cell lines of various species and causes cell death. A human rhabdomyosarcoma cell line, RD-4, infected with BHV-4 produced low levels of early and late viral RNAs and infectious virus, but exhibited no cytopathic effect. Using a recombinant BHV-4 containing a neomycin-resistance gene, we established RD-4-derived cell lines persistently infected with BHV-4. The viral genome in these cells was predominantly circular. Because of drug selection, every cell contained a viral genome. In addition, all cells stained with a BHV-4-specific antiserum. Therefore, these cell lines are not carrier cultures. These cells produced infectious virus at all passages tested. Even though cells were selected and maintained at a concentration of geneticin at least 2·5 times that necessary to kill uninfected RD-4 cells, selected cells contained only approximately one viral genome per diploid host cell genome. Persistently infected cells grew more slowly than uninfected cells, even in the absence of drug. The slower growth of these cells suggests that any growth advantage conferred by multiple copies of the neomycin-gene-carrying viral genome might be offset by the detrimental effects of viral gene expression. This situation contrasts with other gammaherpesviruses, which are able to growth-transform cells.

scholarly journals Integration of bovine herpesvirus 4 genome into cultured persistently infected host cell genome

2010 ◽  
Vol 7 (1) ◽  
Author(s):  
Gaetano Donofrio ◽  
Antonio Capocefalo ◽  
Valentina Franceschi ◽  
Lisa De Lorenzi ◽  
Vicky van Santen ◽  
...  
Keyword(s):  
Host Cell ◽  
Bovine Herpesvirus ◽  
Infected Host ◽  
Persistently Infected ◽  
Infected Host Cell ◽  
Bovine Herpesvirus 4 ◽  
Cell Genome ◽  
Herpesvirus 4

Time Series Analysis of Transmesothelial Invasion by Endometrial Stromal and Epithelial Cells Using Three-dimensional Confocal Microscopy

2001 ◽  
Vol 7 (S2) ◽  
pp. 580-581
Author(s):  
CA Witz ◽  
S Cho ◽  
VE Centonze ◽  
IA Montoya-Rodriguez ◽  
RS Schenken
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Time Course ◽  
Three Dimensional ◽  
Collagen Iv ◽  
Mesothelial Cells ◽  
Endometrial Stromal Cells ◽  
Human Collagen ◽  
Endometrial Epithelial Cells

Using human peritoneal explants, we have previously demonstrated that endometrial stromal cells (ESCs) and endometrial epithelial cells (EECs) attach to intact mesothelium. Attachment occurs within one hour and mesothelial invasion occurs within 18 hours (Figure 1). We have also demonstrated that, in vivo, the mesothelium overlies a continuous layer of collagen IV (Col IV).More recently we have used CLSM, to study the mechanism and time course of ESC and EEC attachment and invasion through mesothelial monolayers. in these studies, CellTracker® dyes were used to label cells. Mesothelial cells were labeled with chloromethylbenzoylaminotetramethylrhodamine (CellTracker Orange). Mesothelial cells were then plated on human collagen IV coated, laser etched coverslips. Mesothelial cells were cultured to subconfluence. ESCs and EECs, labeled with chloromethylfluorscein diacetate (CellTracker Green) were plated on the mesothelial monolayers. Cultures were examined at 1, 6, 12 and 24 hours with simultaneous differential interference contrast and CLSM.

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