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

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.

Download Full-text

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

Journal of General Virology ◽

10.1099/0022-1317-82-5-1181 ◽

2001 ◽

Vol 82 (5) ◽

pp. 1181-1185 ◽

Cited By ~ 34

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.

Download Full-text

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

Microbiology ◽

10.1099/0022-1317-81-7-1807 ◽

2000 ◽

Vol 81 (7) ◽

pp. 1807-1814 ◽

Cited By ~ 16

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.

Download Full-text

Macrophage Activation Redirects Yersinia-Infected Host Cell Death from Apoptosis to Caspase-1-Dependent Pyroptosis

PLoS Pathogens ◽

10.1371/journal.ppat.0030161 ◽

2007 ◽

Vol 3 (11) ◽

pp. e161 ◽

Cited By ~ 133

Author(s):

Tessa Bergsbaken ◽

Brad T Cookson

Keyword(s):

Cell Death ◽

Host Cell ◽

Macrophage Activation ◽

Infected Host ◽

Infected Host Cell ◽

Caspase 1 ◽

Host Cell Death

Download Full-text

Transbilayer phospholipid asymmetry in Plasmodium knowlesi-infected host cell membrane

Science ◽

10.1126/science.7233198 ◽

1981 ◽

Vol 212 (4498) ◽

pp. 1047-1049 ◽

Cited By ~ 51

Author(s):

C. Gupta ◽

G. Mishra

Keyword(s):

Cell Membrane ◽

Host Cell ◽

Plasmodium Knowlesi ◽

Infected Host ◽

Host Cell Membrane ◽

Infected Host Cell ◽

Phospholipid Asymmetry

Download Full-text

Neurological disorder in cattle associated with bovine herpesvirus 4

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/s0102-09352011000400006 ◽

2011 ◽

Vol 63 (4) ◽

pp. 828-835 ◽

Cited By ~ 5

Author(s):

E.A. Costa ◽

A.C. Vasconcelos ◽

M.R.Q. Bomfim ◽

H.B. Amorim ◽

G.B.L. Lima ◽

...

Keyword(s):

Nested Pcr ◽

Clinical Course ◽

Bovine Herpesvirus ◽

Pcr Assay ◽

Bovine Herpesvirus 1 ◽

Herpesvirus Type ◽

Neurological Signs ◽

Bovine Herpesvirus 4 ◽

Herpesvirus 1 ◽

Herpesvirus 4

A nested PCR assay was used to diagnose bovine encephalitis through herpesviruses including bovine herpesvirus 5 (BHV-5), bovine herpesvirus 1 (BHV-1), Aujeszky's disease virus (SHV-1), and ovine herpesvirus 2 (OHV-2) in 14 fragments of central nervous system (CNS) from cattle that died with neurological signs. In addition, as some samples of bovine herpesvirus type 4 (BHV-4) have been isolated from neural tissue, it was also tested by nested PCR. The cases of encephalitis occurred in isolation at different times of the year and did not present any seasonality. The duration of the clinical course ranged between 1 to 15 days, and in 64.3% of the cases it manifested between 1 to 2 days. The most frequently observed neurological signs were ataxia, recumbency, unsteadiness and inability to stand, opisthotonus, paddling movements, nystagmus and ptyalism. In the nested assay, there was no evidence of: BHV-1, SHV-1 or OHV-2 in the DNA obtained from the CNS in any of the samples. But the presence of BHV-4 was found in all fragments of the CNS in cattle which died presenting neurological signs. Moreover, BHV-5 was found in association with BHV-4 in two of these samples.

Download Full-text

Dot Immunobinding Assay for Detection of Bovine Herpesvirus 4 Antibodies in Rabbits

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879901100305 ◽

1999 ◽

Vol 11 (3) ◽

pp. 237-239 ◽

Cited By ~ 3

Author(s):

Mohamed Essmail ◽

Dale Baker ◽

James Collins ◽

Sue Vandewoude ◽

Mohamed Salman ◽

...

Keyword(s):

Bovine Herpesvirus ◽

Bovine Herpesvirus 4 ◽

Herpesvirus 4

Download Full-text

Entropic Forces Drive Clustering and Spatial Localization of Influenza A M2 During Viral Budding

10.1101/291120 ◽

2018 ◽

Author(s):

Jesper J. Madsen ◽

John M. A. Grime ◽

Jeremy S. Rossman ◽

Gregory A. Voth

Keyword(s):

Host Cell ◽

Influenza A ◽

Spatial Localization ◽

Membrane Curvature ◽

Infected Host ◽

Membrane Remodeling ◽

Infected Host Cell ◽

Viral Budding ◽

Virion Release ◽

Liquid Ordered

ABSTRACTThe influenza A matrix 2 (M2) transmembrane protein facilitates virion release from the infected host cell. In particular, M2 plays a role in the induction of membrane curvature and/or in the scission process whereby the envelope is cut upon virion release. Here we show using coarse-grained computer simulations that various M2 assembly geometries emerge due to an entropic driving force, resulting in compact clusters or linearly extended aggregates as a direct consequence of the lateral membrane stresses. Conditions under which these protein assemblies will cause the lipid membrane to curve are explored and we predict that a critical cluster size is required for this to happen. We go on to demonstrate that under the stress conditions taking place in the cellular membrane as it undergoes large-scale membrane remodeling, the M2 protein will in principle be able to both contribute to curvature induction and sense curvature in order to line up in manifolds where local membrane line tension is high. M2 is found to exhibit linactant behavior in liquid-disordered/liquid-ordered phase-separated lipid mixtures and to be excluded from the liquid-ordered phase, in near-quantitative agreement with experimental observations. Our findings support a role for M2 in membrane remodeling during influenza viral budding both as an inducer and a sensor of membrane curvature, and they suggest a mechanism by which localization of M2 can occur as the virion assembles and releases from the host cell, independent of how the membrane curvature is produced.SIGNIFICANCE STATEMENTFor influenza virus to release from the infected host cell, controlled viral budding must finalize with membrane scission of the viral envelope. Curiously, influenza carries its own protein, M2, which can sever the membrane of the constricted budding neck. Here we elucidate the physical mechanism of clustering and spatial localization of the M2 scission proteins through a combined computational and experimental approach. Our results provide fundamental insights into how M2 clustering and localization interplays with membrane curvature, membrane lateral stresses, and lipid bilayer phase behavior during viral budding in order to contribute to virion release.

Download Full-text