RESUMEN
Koi herpesvirus disease (KHVD) is an emerging and highly contagious viral disease of koi and common carp (Cyprinus carpio), causing mass mortalities and huge economic losses to the carp aquaculture industry. The disease has spread rapidly to 28 countries worldwide. However, mechanisms of koi herpesvirus (species Cyprinid herpesvirus 3; CyHV-3) transmission remain unclear. A potential experimental model of CyHV-3 infection in carp was used to characterise CyHV-3 in different phases of infection and to demonstrate that CyHV-3 persists in survivor fish and has the capacity to reactivate and transmit the disease to healthy fish. During acute infection, which occurred when fish were maintained at 22°C, viral genes were abundantly expressed and infectious virus was produced in association with tissue damage, clinical disease and mortality. In fish maintained at a lower temperature (11°C), viral DNA was present but viral gene expression was absent or greatly restricted, infectious virus was not recovered and there was no evidence of disease. Productive replication was re-initiated following an increase in water temperature to 22°C, resulting in 45% mortality. Shedding of reactivated virus killed 75% of cohabitating naïve fish, suggesting a potential risk for disease transmission.
Asunto(s)
Carpas/virología , Infecciones por Herpesviridae/veterinaria , Herpesviridae/fisiología , Replicación Viral/efectos de la radiación , Animales , Portador Sano/veterinaria , Portador Sano/virología , Herpesviridae/efectos de la radiación , Infecciones por Herpesviridae/transmisión , Infecciones por Herpesviridae/virología , Análisis de Supervivencia , Temperatura , Activación Viral , Esparcimiento de VirusRESUMEN
Nucleotide-binding oligomerization domain 2 (NOD2) is an important innate immune sensor of bacterial pathogens. Its induction results in activation of the classic NF-κB pathway and alternative pathways including type I IFN and autophagy. Although the importance of NOD2 in recognizing RNA viruses has recently been identified, its role in sensing DNA viruses has not been studied. We report that infection with human cytomegalovirus (HCMV) results in significant induction of NOD2 expression, beginning as early as 2 hours post infection and increasing steadily 24 hours post infection and afterwards. Infection with human herpesvirus 1 and 2 does not induce NOD2 expression. While the HCMV-encoded glycoprotein B is not required for NOD2 induction, a replication competent virion is necessary. Lentivirus-based NOD2 knockdown in human foreskin fibroblasts (HFFs) and U373 glioma cells leads to enhanced HCMV replication along with decreased levels of interferon beta (IFN-ß) and the pro-inflammatory cytokine, IL8. NOD2 induction in HCMV-infected cells activates downstream NF-κB and interferon pathways supported by reduced nuclear localization of NF-κB and pIRF3 in NOD2 knockdown HFFs. Stable overexpression of NOD2 in HFFs restricts HCMV replication in association with increased levels of IFN-ß and IL8. Similarly, transient overexpression of NOD2 in U373 cells or its downstream kinase, RIPK2, results in decreased HCMV replication and enhanced cytokine responses. However, overexpression of a mutant NOD2, 3020insC, associated with severe Crohn's disease, results in enhanced HCMV replication and decreased levels of IFN-ß in U373 cells. These results show for the first time that NOD2 plays a significant role in HCMV replication and may provide a model for studies of HCMV recognition by the host cell and HCMV colitis in Crohn's disease.
Asunto(s)
Citomegalovirus/fisiología , Inmunidad Innata , Proteína Adaptadora de Señalización NOD2/metabolismo , Replicación Viral/fisiología , Línea Celular , Citocinas/metabolismo , Citomegalovirus/efectos de la radiación , Infecciones por Citomegalovirus/inmunología , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Fibroblastos/virología , Técnicas de Silenciamiento del Gen , Herpesviridae/fisiología , Herpesviridae/efectos de la radiación , Humanos , Inmunidad Innata/efectos de la radiación , Mediadores de Inflamación/metabolismo , Interferones/metabolismo , Cinética , Masculino , Proteínas Mutantes/metabolismo , FN-kappa B/metabolismo , Proteína Adaptadora de Señalización NOD2/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Rayos Ultravioleta , Proteínas del Envoltorio Viral/metabolismo , Inactivación de Virus/efectos de la radiación , Replicación Viral/efectos de la radiaciónRESUMEN
Cyprinid herpesvirus-3 (CyHV-3) is a member of the Alloherpesviridae, in the order Herpesvirales. It causes a fatal disease in carp and koi fish. The disease is seasonal and is active when water temperatures ranges from 18 to 28 °C. Little is known about how and where the virus is preserved between the permissive seasons. The hallmark of the herpesviruses is their ability to become latent, persisting in the host in an apparently inactive state for varying periods of time. Hence, it could be expected that CyHV-3 enter a latent period. CyHV-3 has so far been shown to persist in fish maintained under restrictive temperatures, while shifting the fish to permissive conditions reactivates the virus. Previously, we demonstrated that cultured cells infected with CyHV-3 at 22 °C and subsequently transferred to a restrictive temperature of 30 °C preserve the virus for 30 days. The present report shows that cultured carp cells maintained and exposed to CyHV-3 at 30 °C are abortively infected; that is, autonomous viral DNA synthesis is hampered and the viral genome is not multiplied. Under these conditions, 91 of the 156 viral annotated ORFs were initially transcribed. These transcripts were down-regulated and gradually shut off over 18 days post-infection, while two viral transcripts encoded by ORFs 114 and 115 were preserved in the infected cells for 18 days p.i. These experiments, carried out in cultured cells, suggest that fish could be infected at a high non-permissive temperature and harbor the viral genome without producing viral particles.
Asunto(s)
Carpas/virología , Regulación hacia Abajo , Regulación Viral de la Expresión Génica/efectos de la radiación , Herpesviridae/fisiología , Herpesviridae/efectos de la radiación , Latencia del Virus/efectos de la radiación , Animales , Células Cultivadas , Herpesviridae/genética , CalorRESUMEN
The transmission of human immunodeficiency virus (HIV-1) and other enveloped virus by blood transfusion is a major concern. Photosensitive dyes such as hematoporphyrin derivative (HPD), dihematoporphyrin ether (DHE), benzoporphyrin derivatives (BPD), extended ring porphyrins, sapphyrins and texaphyrins, and various cyanines were used with viral cultures to test the feasibility of using those light-excitable dyes to kill virus. A photodynamic flow cell was used to irradiate viral suspensions or viral infected cells in culture media or in whole blood. Herpes virus (HSV-1) was used to screen compounds. Effective compounds were subsequently tested for their ability to kill HIV-1, CMV, and SIV in culture medium and in blood and proved to effectively kill free virus and infected cells at significant viremias. Irradiation was achieved with a filtered xenon light source and/or tunable dye laser. Concentrations of dyes at 10 times viral kill dose were irradiated in blood which was tested for damage to erythrocytes (RBC), platelets, and blood proteins. No damage to RBC, complement factors, and immunoglobulins was evident immediately after photodynamic treatment. Platelet condition is minimally modified with time. Photodynamic treatment of blood appears to be a feasible means of eradicating virus and some protozoans from blood.
Asunto(s)
Colorantes/farmacología , Fármacos Sensibilizantes a Radiaciones/farmacología , Rayos Ultravioleta , Virus/efectos de los fármacos , Sangre/efectos de los fármacos , Sangre/microbiología , Sangre/efectos de la radiación , Células Sanguíneas/efectos de los fármacos , Células Sanguíneas/efectos de la radiación , Membrana Celular/efectos de los fármacos , Membrana Celular/efectos de la radiación , Colorantes/efectos de la radiación , Proteínas del Sistema Complemento/efectos de los fármacos , Proteínas del Sistema Complemento/efectos de la radiación , Medios de Cultivo , VIH-1/efectos de los fármacos , VIH-1/fisiología , VIH-1/efectos de la radiación , Herpesviridae/efectos de los fármacos , Herpesviridae/fisiología , Herpesviridae/efectos de la radiación , Humanos , Rayos Láser , Fotoquímica/instrumentación , Fármacos Sensibilizantes a Radiaciones/efectos de la radiación , Virus de la Inmunodeficiencia de los Simios/efectos de los fármacos , Virus de la Inmunodeficiencia de los Simios/fisiología , Virus de la Inmunodeficiencia de los Simios/efectos de la radiación , Fenómenos Fisiológicos de los Virus , Replicación Viral/efectos de los fármacos , Replicación Viral/efectos de la radiación , Virus/efectos de la radiaciónRESUMEN
The present studies were undertaken to determine if viral particles can be delivered across the rat blood-brain barrier (BBB). Osmotic BBB modification with intracarotid mannitol (25%) was immediately followed by bolus intracarotid administration of 0.5 ml purified, ultraviolet-inactivated, herpes simplex virus type 1 endogenously labeled with 35S-labeled methionine (2.0 x 10(6) cpm, approximately 5 x 10(8) plaque-forming units/ml). After 60 minutes, intravascular virus was cleared by saline perfusion and the animals were sacrificed. A marked increase (fourfold, p less than or equal to 0.02) in radioactivity was observed in the ipsilateral brain hemisphere when compared to control animals without barrier modification. Administration of intravenous virus immediately after BBB modification displayed no difference in delivery when compared to intracarotid saline-infused controls (without BBB modification) suggesting the importance of a first-pass phenomenon. There were no significant differences in serum concentrations among intracarotid or intravenous groups. These preliminary studies suggest the possibility of delivering viral particles across the BBB with osmotic disruption, which may permit delivery of genetic material in replication-defective viral vectors in the feline model of GM2-gangliosidosis.
Asunto(s)
Barrera Hematoencefálica , Herpesviridae/efectos de la radiación , Rayos Ultravioleta , Animales , Barrera Hematoencefálica/efectos de los fármacos , Femenino , Vectores Genéticos , Herpesviridae/fisiología , Manitol/farmacología , Ósmosis , Ratas , Activación ViralRESUMEN
The effect of temperature, pH, salinity and ultraviolet irradiation on the infectivity of Herpes Channel Catfish Virus (HCCV) has been determined. It has been shown that the pH and the culture medium is an important factor in maintaining virus infectivity. The temperatures higher than 4 degrees C rapidly inactivate the virus while this one can be grown successfully after several months of storage at -75 degrees C and at -20 degrees C. In common with other herpesviruses, HCCV is quickly inactivated by UV irradiation. The signification of these findings on the mode of viral infection and of viral harvesting, disinfection measures and marine farming of Ictalurids is important.
Asunto(s)
Herpesviridae/fisiología , Animales , Peces , Herpesviridae/efectos de la radiación , Concentración de Iones de Hidrógeno , Concentración Osmolar , Temperatura , Rayos UltravioletaRESUMEN
The small-plaque effect occurs with a wide range of herpesviruses following irradiation with ultraviolet light. The 37 per cent survival (D37) values, or dose required for one lethal hit (e-1), for herpes simplex, pseudorabies and pigeon herpesviruses in different cells indicate a broad spectrum of host-cell repair capacity. Other DNA-containing viruses such as SV40 and adenoviruses, which also replicate in the cell nucleus, show the small-plaque effect. Ionizing irradiation of herpes simplex virus type 1 (HSV-1) showed but little reduction in plaque-size.
Asunto(s)
Adenovirus Humanos/efectos de la radiación , Electrones , Herpesviridae/efectos de la radiación , Radiación Ionizante , Virus 40 de los Simios/efectos de la radiación , Rayos Ultravioleta , Replicación Viral/efectos de la radiación , Adenovirus Humanos/crecimiento & desarrollo , Animales , Línea Celular , Efecto Citopatogénico Viral/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Herpesviridae/crecimiento & desarrollo , Humanos , Virus 40 de los Simios/crecimiento & desarrolloRESUMEN
Several virion and nonvirion DNAs were tested for the ability to activate endogenous type C virus in BALB/c-derived mouse cells using the calcium precipitation technique. The DNAs from all herpesviruses tested activated xenotropic type C virus synthesis. These included DNAs from herpes simplex virus types 1 and 2, Epstein-Barr virus, human cytomegalovirus, SA8 virus, infectious bovine rhinotracheitis virus, pseudorabies virus, and herpes saimiri virus (M-DNA). In contrast, DNAs from vaccinia virus, simian virus 40, primate cells, bacteria, mycoplasma, and salmon sperm showed no ability to activate type C virus when tested under the same conditions. Several herpesviruses and vaccinia virus, which were highly infectious for the BALB/c cells used, were tested for their ability to activate type C virus after UV irradiation. All herpesviruses tested were positive, while vaccinia virus was negative. Unirradiated simian virus 40 also showed no ability to activate type C virus. Activation of type C virus by DNA from herpes simplex virus was observed after shearing or sonication of the DNA to an average size of 3 x 10(6) daltons, but was not observed with DNA sonicated to an average size of 1 x 10(6) daltons. Alkali denaturation of DNA from herpes simplex virus or treatment with DNase, but not RNase, destroyed its ability to activate type C virus, as did crosslinking of the DNA with 4,5',8-trimethylpsoralen (psoralen) and light.
Asunto(s)
ADN Viral/metabolismo , Herpesviridae/metabolismo , Retroviridae/crecimiento & desarrollo , Replicación Viral , Línea Celular , ADN Viral/farmacología , Relación Dosis-Respuesta a Droga , Herpesviridae/efectos de la radiación , Peso Molecular , Replicación Viral/efectos de los fármacosRESUMEN
Chicken embryo fibroblasts infected with the nononcogenic herpesvirus of turkeys (HVT) displayed an increased rate of glucose uptake, a pronounced alteration of the pH of the medium, and an increased production of lactic acid when compared to mock-infected cultures. Objective estimates of cytopathology (quantifiable neutral red uptake and cell protein determination) showed that cell deterioration was a slow process in HVT-infected cells when compared to infection by herpes simplex virus. Experiments with irradiated host cells demonstrated that HVT required functional cell DNA for replication. The inactivation of the necessary host cell function displayed multihit kinetics. In agreement with data on other herpesviruses, HVT damaged by UV light could be photoreactivated in chick cells. The results indicate that HVT shares biologic properties in common with other herpes and transforming viruses.
Asunto(s)
Glucosa/metabolismo , Herpesviridae , Pavos/microbiología , Animales , Células Cultivadas , Embrión de Pollo , Medios de Cultivo , Efecto Citopatogénico Viral , Herpesviridae/crecimiento & desarrollo , Herpesviridae/efectos de la radiación , Concentración de Iones de Hidrógeno , Lactatos/metabolismo , Efectos de la Radiación , Simplexvirus , Rayos Ultravioleta , Replicación ViralAsunto(s)
Herpesviridae , Interferones/biosíntesis , Seudorrabia/microbiología , Animales , Células Cultivadas , Efecto Citopatogénico Viral , Fibroblastos , Herpesviridae/crecimiento & desarrollo , Herpesviridae/patogenicidad , Herpesviridae/efectos de la radiación , Calor , Efectos de la Radiación , Rayos Ultravioleta , Virulencia , Replicación ViralAsunto(s)
Encéfalo/microbiología , Herpesviridae/crecimiento & desarrollo , Animales , Encéfalo/citología , Células Cultivadas , Efecto Citopatogénico Viral , Cobayas , Herpesviridae/efectos de la radiación , Cuerpos de Inclusión Viral , Ratones , Microscopía Electrónica , Efectos de la Radiación , Especificidad de la Especie , Rayos Ultravioleta , Replicación ViralRESUMEN
Ultraviolet light-inactivated frog virus 3 is efficiently photoreactivated by chick embryo cells. A cellular enzyme is presumably responsible for this repair of viral deoxyribonucleic acid, for the phenomenon is insensitive to an inhibitor of protein synthesis and is not seen in mammalian cells that are known to lack photoreactivating enzyme. Since frog virus 3 is a cytoplasmic virus, functionally significant amounts of photoreactivating enzyme are probably present in the cytoplasm of chick embryo cells.