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1.
PLoS Pathog ; 18(10): e1010905, 2022 10.
Artículo en Inglés | MEDLINE | ID: mdl-36240255

RESUMEN

Viral interference is a process where infection with one virus prevents a subsequent infection with the same or a different virus. This is believed to limit superinfection, promote viral genome stability, and protect the host from overwhelming infection. Mechanisms of viral interference have been extensively studied in plants, but remain poorly understood in vertebrates. We demonstrate that infection with infectious salmon anaemia virus (ISAV) strongly reduces homologous viral attachment to the Atlantic salmon, Salmo salar L. vascular surface. A generalised loss of ISAV binding was observed after infection with both high-virulent and low-virulent ISAV isolates, but with different kinetics. The loss of ISAV binding was accompanied by an increased susceptibility to sialidase, suggesting a loss of the vascular 4-O-sialyl-acetylation that mediates ISAV attachment and simultaneously protects the sialic acid from cleavage. Moreover, the ISAV binding capacity of cultured cells dramatically declined 3 days after ISAV infection, accompanied by reduced cellular permissiveness to infection with a second antigenically distinct isolate. In contrast, neither infection with infectious haematopoietic necrosis virus nor stimulation with the viral mimetic poly I:C restricted subsequent cellular ISAV attachment, revealing an ISAV-specific mechanism rather than a general cellular antiviral response. Our study demonstrates homologous ISAV attachment interference by de-acetylation of sialic acids on the vascular surface. This is the first time the kinetics of viral receptor destruction have been mapped throughout the full course of an infection, and the first report of homologous attachment interference by the loss of a vascular viral receptor. Little is known about the biological functions of vascular O-sialyl-acetylation. Our findings raise the question of whether this vascular surface modulation could be linked to the breakdown of central vascular functions that characterises infectious salmon anaemia.


Asunto(s)
Anemia , Enfermedades de los Peces , Isavirus , Infecciones por Orthomyxoviridae , Salmo salar , Animales , Isavirus/genética , Receptores Virales
2.
J Gen Virol ; 98(4): 595-606, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28475029

RESUMEN

The putatively non-virulent subtype of infectious salmon anaemia virus (ISAV), ISAV-HPR0, is proposed to act as a progenitor and reservoir for all virulent ISAVs and thus represent a potential risk factor for the emergence of infectious salmon anaemia (ISA) disease. Here, we provide the first evidence of genetic and functional evolution from an ISAV-HPR0 variant (FO/07/12) to a low-virulent ISAV virus (FO/121/14) in a Faroese Atlantic salmon marine farm. The FO/121/14 virus infection was not associated with specific clinical signs of ISA and was confined to a single net-pen, while various ISAV-HPR0 subtypes were found circulating in most epidemiologically linked marine and freshwater farms. Sequence analysis of all eight segments revealed that the FO/121/14 virus was identical, apart from a substitution in the fusion (F) gene (Q266L) and a deletion in the haemagglutinin-esterase (HE) gene, to the FO/07/12 variant from a freshwater farm, which supplied smolts exclusively to the FO/121/14-positive net-pen. An immersion challenge with the FO/121/14 virus induced a systemic infection in Atlantic salmon associated with a low mortality and mild clinical signs confirming its low pathogenicity. Our results demonstrate that mutations in the F protein and deletions in the highly polymorphic region (HPR) of the HE protein represent a minimum requirement for ISAV to gain virulence and to switch cell tropism from a localized epithelial infection to a systemic endotheliotropic infection. This documents that ISAV-HPR0 represents a reservoir and risk factor for the emergence of ISA disease.


Asunto(s)
Evolución Molecular , Enfermedades de los Peces/virología , Isavirus/genética , Infecciones por Orthomyxoviridae/veterinaria , Animales , Isavirus/clasificación , Isavirus/aislamiento & purificación , Isavirus/patogenicidad , Mutación , Infecciones por Orthomyxoviridae/virología , Filogenia , Salmo salar , Proteínas Virales/genética , Virulencia
3.
J Immunol ; 193(7): 3463-9, 2014 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-25172486

RESUMEN

Previously, our group has shown that the interbranchial lymphoid tissue (ILT) is a distinct structure largely consisting of T cells embedded in a meshwork of epithelial cells, with no direct resemblance to previously described lymphoid tissues. In this study, we aim to focus on the T cell population and the possibility of the ILT being a thymus analog. By characterizing structural responsiveness to Ag challenge, the presence of recombination activating genes, and different T cell-related transcripts, we attempt to further approach the immunological function of the ILT in salmonid gills. In addition to eight healthy individuals, a group of eight infectious salmon anemia virus-challenged fish were included to observe T cell responses related to infection. The results showed reduced size of ILT in the infected group, no expression of RAG-1 and -2, and a high degree of T cell diversity within the ILT. Taking into account that the ILT can be regarded as a strategically located T cell reservoir and possibly an evolutionary forerunner of mammalian MALTs right at the border to the external environment, the alteration in transcription observed may likely represent a shift in the T cell population to optimize local gill defense mechanisms.


Asunto(s)
Regulación de la Expresión Génica/inmunología , Branquias/inmunología , Tejido Linfoide/inmunología , Salmo salar/inmunología , Linfocitos T/inmunología , Transcripción Genética/inmunología , Animales , Proteínas de Unión al ADN/inmunología , Branquias/citología , Proteínas de Homeodominio/inmunología , Tejido Linfoide/citología , Linfocitos T/citología
5.
Vet Res ; 46: 120, 2015 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-26490835

RESUMEN

All viruses infecting fish must cross the surface mucosal barrier to successfully enter a host. Infectious salmon anaemia virus (ISAV), the causative agent of the economically important infectious salmon anaemia (ISA) in Atlantic salmon, Salmo salar L., has been shown to use the gills as its entry point. However, other entry ports have not been investigated despite the expression of virus receptors on the surface of epithelial cells in the skin, the gastrointestinal (GI) tract and the conjunctiva. Here we investigate the ISAV mucosal infection in Atlantic salmon after experimental immersion (bath) challenge and in farmed fish collected from a confirmed outbreak of ISA in Norway. We show for the first time evidence of early replication in several mucosal surfaces in addition to the gills, including the pectoral fin, skin and GI tract suggesting several potential entry points for the virus. Initially, the infection is localized and primarily infecting epithelial cells, however at later stages it becomes systemic, infecting the endothelial cells lining the circulatory system. Viruses of low and high virulence used in the challenge revealed possible variation in virus progression during infection at the mucosal surfaces.


Asunto(s)
Enfermedades de los Peces/virología , Isavirus/fisiología , Membrana Mucosa/virología , Infecciones por Orthomyxoviridae/veterinaria , Salmo salar , Animales , Acuicultura , Noruega , Infecciones por Orthomyxoviridae/virología , Replicación Viral
6.
J Gen Virol ; 95(Pt 5): 1015-1024, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24486627

RESUMEN

Since the discovery of a non-virulent infectious salmon anaemia virus (ISAV) HPR0 variant, many studies have speculated on the functional role of deletions within the highly polymorphic region (HPR) of genomic segment 6, which codes for the haemagglutinin-esterase (HE) protein. To address this issue, mutant HE proteins with deletions in their HPR were generated from the Scottish HPR0 template (NWM10) and fusion-inducing activity was measured using lipid (octadecyl rhodamine B) and content mixing assays (firefly luciferase). Segment six HPR was found to have a strong influence on ISAV fusion, and deletions in this near-membrane region predominantly increased the fusion-inducing ability of the resulting HE proteins. The position and length of the HPR deletions were not significant factors, suggesting that they may affect fusion non-specifically. In comparison, the amino acid composition of the associated fusion (F) protein was a more crucial criterion. Antibody co-patching and confocal fluorescence demonstrated that the HE and F proteins were highly co-localized, forming defined clusters on the cell surface post-transfection. The binding of erythrocyte ghosts on the attachment protein caused a reduction in the percentage of co-localization, suggesting that ISAV fusion might be triggered through physical separation of the F and HE proteins. In this process, HPR deletion appeared to modulate and reduce the strength of interaction between the two glycoproteins, causing more F protein to be released and activated. This work provides a first insight into the mechanism of virulence acquisition through HPR deletion, with fusion enhancement acting as a major contributing factor.


Asunto(s)
Hemaglutininas Virales/genética , Hemaglutininas Virales/metabolismo , Isavirus/fisiología , Eliminación de Secuencia , Proteínas Virales de Fusión/metabolismo , Internalización del Virus , Animales , Células Cultivadas , Isavirus/genética , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Unión Proteica , Salmón , Proteínas Virales de Fusión/genética
7.
Glycoconj J ; 31(4): 327-35, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24833039

RESUMEN

Sialic acids are located at the terminal branches of the cell glycocalyx and secreted glycan molecules. O-Acetylation is an important modification of the sialic acids, however very few studies have demonstrated the in situ distribution of the O-Acetylated sialic acids. Here the distribution of glycoprotein bound 4-O-Acetylated sialic acids (4-O-Ac sias) in vertebrates was determined using a novel virus histochemistry assay. The 4-O-Ac sias were found in the circulatory system, i.e. on the surface of endothelial cells and RBCs, of several vertebrate species, though most frequently in the cartilaginous fish (class Chondrichthyes) and the bony fish (class Osteichthyes). The O-Acetylated sialic acid was detected in 64 % of the examined fish species. Even though the sialic acid was found less commonly in higher vertebrates, it was found at the same location in the positive species. The general significance of this endothelial labelling pattern distribution is discussed. The seemingly conserved local position through the evolution of the vertebrates, suggests an evolutionary advantage of this sialic acid modification.


Asunto(s)
Glicoproteínas/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Acetilación , Animales , Células Endoteliales/metabolismo , Eritrocitos/metabolismo , Glicoproteínas/genética , Especificidad de la Especie , Vertebrados
8.
Vet Res ; 45: 83, 2014 Aug 21.
Artículo en Inglés | MEDLINE | ID: mdl-25143055

RESUMEN

Observations from the field and experimental evidence suggest that different strains of infectious salmon anaemia virus (ISAV) can induce disease of varying severity in Atlantic salmon. Variation in host mortality and dissemination of ISAV isolates with high and low virulence was investigated using immersion challenge; from which mortality, pathological, immunohistochemical and preliminary molecular results have been previously published. Here, real-time RT-PCR analysis and statistical modelling have been used to further investigate variation in virus load and the response of four select immune genes. Expression of type I and II interferon (IFN), Mx and γIFN induced protein (γIP) to high and low pathogenic virus infection were examined in gill, heart and anterior kidney. In addition, a novel RNA species-specific assay targeting individual RNA types was used to investigate the separate viral processes of transcription and replication. Unexpectedly, the low virulent ISAV (LVI) replicated and transcribed more rapidly in the gills compared to the highly virulent virus (HVI). Subsequently LVI was able to disseminate to the internal organs more quickly and induced a more rapid systemic immune response in the host that may have offered some protection. Contrary to this, HVI initially progressed more slowly in the gills resulting in a slower generalised infection. However HVI ultimately reached a higher viral load and induced a greater mortality.


Asunto(s)
Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Proteínas de Peces/genética , Isavirus/fisiología , Isavirus/patogenicidad , Infecciones por Orthomyxoviridae/veterinaria , Salmo salar , Animales , Enfermedades de los Peces/mortalidad , Proteínas de Peces/metabolismo , Inmunohistoquímica/veterinaria , Isavirus/inmunología , Modelos Teóricos , Datos de Secuencia Molecular , Especificidad de Órganos , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/mortalidad , Infecciones por Orthomyxoviridae/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Análisis de Secuencia de ADN/veterinaria , Carga Viral/veterinaria , Virulencia , Replicación Viral/fisiología
9.
J Virol ; 86(19): 10571-8, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22811536

RESUMEN

Infectious salmon anemia (ISA) is a World Organization for Animal Health (OIE)-listed disease of farmed Atlantic salmon, characterized by slowly developing anemia and circulatory disturbances. The disease is caused by ISA virus (ISAV) in the Orthomyxoviridae family; hence, it is related to influenza. Here we explore the pathogenesis of ISA by focusing on virus tropism, receptor tissue distribution, and pathological changes in experimentally and naturally infected Atlantic salmon. Using immunohistochemistry on ISAV-infected Atlantic salmon tissues with antibody to viral nucleoprotein, endotheliotropism was demonstrated. Endothelial cells lining the circulatory system were found to be infected, seemingly noncytolytic, and without vasculitis. No virus could be found in necrotic parenchymal cells. From endothelium, the virus budded apically and adsorbed to red blood cells (RBCs). No infection or replication within RBCs was detected, but hemophagocytosis was observed, possibly contributing to the severe anemia in fish with this disease. Similarly to what has been done in studies of influenza, we examined the pattern of virus attachment by using ISAV as a probe. Here we detected the preferred receptor of ISAV, 4-O-acetylated sialic acid (Neu4,5Ac(2)). To our knowledge, this is the first report demonstrating the in situ distribution of this sialic acid derivate. The pattern of virus attachment mirrored closely the distribution of infection, showing that the virus receptor is important for cell tropism, as well as for adsorption to RBCs.


Asunto(s)
Eritrocitos/virología , Enfermedades de los Peces/virología , Regulación de la Expresión Génica , Infecciones por Orthomyxoviridae/metabolismo , Adsorción , Animales , Inmunohistoquímica/métodos , Leucocitos/citología , Microscopía Fluorescente/métodos , Ácido N-Acetilneuramínico/química , Fagocitosis , Salmo salar , Distribución Tisular , Tropismo Viral
10.
J Anat ; 222(5): 547-57, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23439106

RESUMEN

Endothelial cells (ECs) line the luminal surfaces of the cardiovascular system and play an important role in cardiovascular functions such as regulation of haemostasis and vasomotor tone. A number of fish and mammalian viruses target these cells in the course of their infection. Infectious salmon anaemia virus (ISAV) attacks ECs and red blood cells (RBCs) of farmed Atlantic salmon (Salmo salar L.), producing the severe disease of infectious salmon anaemia (ISA). The investigation of ISA has up to now been hampered by the lack of a functional marker for ECs in Atlantic salmon in situ. In this study, we report the characterisation and use of a novel monoclonal antibody (MAb) detecting Atlantic salmon ECs (e.g. vessel endothelium, endocardial cells and scavenger ECs) and RBCs. The antibody can be used with immunohistochemistry, IFAT and on Western blots. It appears that the epitope recognised by the antibody is associated with the ISAV cellular receptor. Besides being a tool to identify ECs in situ, it could be useful in further studies of the pathogenicity of ISA. Finally, the detection of an epitope shared by ECs and RBCs agrees with recent findings that these cells share a common origin, thus the MAb can potentially be used to study the ontogeny of these cells in Atlantic salmon.


Asunto(s)
Anticuerpos Monoclonales , Células Endoteliales/inmunología , Eritrocitos/inmunología , Enfermedades de los Peces/inmunología , Isavirus/inmunología , Receptores Virales/inmunología , Salmo salar/virología , Animales , Células Endoteliales/citología , Eritrocitos/citología , Enfermedades de los Peces/virología , Inmunohistoquímica
11.
Virol J ; 10: 5, 2013 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-23282149

RESUMEN

Infectious salmon anaemia virus (ISAV), a member of the Orthomyxoviridae family, infects and causes disease in farmed Atlantic salmon (Salmo salar L.). Previous studies have shown Atlantic salmon endothelial cells to be the primary targets of ISAV infection. However, it is not known if cells other than endothelial cells play a role in ISAV tropism. To further assess cell tropism, we examined ISAV infection of Atlantic salmon gill epithelial cells in vivo and in vitro. We demonstrated the susceptibility of epithelial cells to ISAV infection. On comparison of primary gill epithelial cell cultures with ISAV permissive fish cell cultures, we found the virus yield in primary gill epithelial cells to be comparable with that of salmon head kidney (SHK)-1 cells, but lower than TO or Atlantic salmon kidney (ASK)-II cells. Light and transmission electron microscopy (TEM) revealed that the primary gill cells possessed characteristics consistent with epithelial cells. Virus histochemistry showed that gill epithelial cells expressed 4-O-acetylated sialic acid which is recognized as the ISAV receptor. To the best of our knowledge, this is the first demonstration of ISAV infection in Atlantic salmon primary gill epithelial cells. This study thus broadens our understanding of cell tropism and transmission of ISAV in Atlantic salmon.


Asunto(s)
Células Epiteliales/virología , Branquias/virología , Isavirus/patogenicidad , Salmo salar/virología , Tropismo Viral , Animales , Línea Celular , Histocitoquímica , Inmunohistoquímica , Isavirus/crecimiento & desarrollo , Microscopía Electrónica , Receptores Virales/análisis , Ácidos Siálicos/análisis , Cultivo de Virus
12.
Vet Res ; 44: 107, 2013 Nov 13.
Artículo en Inglés | MEDLINE | ID: mdl-24219276

RESUMEN

It is widely accepted that melanin formation may play an immunologic role in invertebrates and ectothermic vertebrates. In farmed Atlantic salmon, cardiomyopathy syndrome (CMS) is a common viral disease associated with severe cardiac inflammation that may be accompanied by heavy melanisation of the heart. By the use of histology, laser capture microdissection and transcription analysis of tyrosinase genes, we here show that this melanisation is linked to de novo melanogenesis by melanomacrophages, suggesting an active part in the inflammatory reaction. No general systemic activation of the extracutaneous pigmentary system in response to viral infections with affinity to the heart was observed.


Asunto(s)
Enfermedades de los Peces/patología , Melaninas/metabolismo , Miocarditis/veterinaria , Miocardio/patología , Infecciones por Virus ARN/veterinaria , Salmo salar , Totiviridae/fisiología , Animales , Enfermedades de los Peces/fisiopatología , Enfermedades de los Peces/virología , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Oxidorreductasas Intramoleculares/genética , Oxidorreductasas Intramoleculares/metabolismo , Captura por Microdisección con Láser/veterinaria , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/metabolismo , Miocarditis/patología , Miocarditis/fisiopatología , Miocarditis/virología , Miocardio/inmunología , Noruega , Infecciones por Virus ARN/patología , Infecciones por Virus ARN/fisiopatología , Infecciones por Virus ARN/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria
13.
Dis Aquat Organ ; 103(1): 35-43, 2013 Mar 13.
Artículo en Inglés | MEDLINE | ID: mdl-23482383

RESUMEN

The prevalence and geographical distribution of the recently described endosymbiont 'Candidatus Branchiomonas cysticola' in Atlantic salmon Salmo salar gill epithelial cell cysts was investigated in seawater-farmed fish suffering proliferative gill inflammation (PGI). To this end, we developed a specific and sensitive real-time PCR assay for detection of the bacterium. 'Ca. B. cysticola' was found to be highly prevalent in Atlantic salmon gills sampled over 7 yr and from 17 geographically distant seawater locations in Norway and Ireland. 'Ca. B. cysticola' was found in significantly greater quantities in fish with large numbers of epitheliocysts, and fluorescence in situ hybridization confirmed its localisation within cysts. 'Ca. Piscichlamydia salmonis', a bacterium previously linked to epitheliocysts, was identified at relatively low levels of infection, apparently independent of epitheliocyst prevalence. These results suggest that 'Ca. B. cysticola' is the main cyst-forming bacterium in seawater-farmed Atlantic salmon in the studied countries. Our results also suggest a relationship between load of 'Ca. B. cysticola' and extent of pathological changes. Taken together with a previously described association between epitheliocyst load and severity of PGI in Norwegian salmon, the results could indicate a role for 'Ca. B. cysticola' in gill diseases such as PGI.


Asunto(s)
Acuicultura/métodos , Infecciones Bacterianas/veterinaria , Betaproteobacteria/aislamiento & purificación , Enfermedades de los Peces/microbiología , Salmo salar , Enfermedades Cutáneas Bacterianas/veterinaria , Animales , Infecciones Bacterianas/epidemiología , Infecciones Bacterianas/microbiología , ADN Bacteriano , Brotes de Enfermedades/veterinaria , Enfermedades de los Peces/epidemiología , Irlanda/epidemiología , Noruega/epidemiología , Prevalencia , Enfermedades Cutáneas Bacterianas/epidemiología , Enfermedades Cutáneas Bacterianas/microbiología
14.
J Virol Methods ; 321: 114791, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37562733

RESUMEN

Infectious salmon anaemia virus (ISAV) can cause severe systemic infection in Atlantic salmon (Salmo salar L.), and a timely diagnosis is critical. Conventional real-time reverse transcription PCR (RT-qPCR) assays target unspliced RNA from either ISAV segment 7 or 8 and provide data on viral load. Here, we evaluate a TaqMan one-step RT-qPCR assay that detects explicitly a spliced messenger RNA (mRNA) of ISAV segment 7, thus providing evidence of active viral transcription. Assay performance was comparable with existing unspliced segment 7 and segment 8 assays. PCR efficiency as evaluated from dilutions of a synthetic DNA fragment was 98 % (R2 = 1.00). The assay also performed well on clinical heart samples with PCR efficiency of 108 % (R2 = 1.00). Finally, evaluation on kidney samples from experimental infection revealed higher levels of active transcription for high-virulent compared to low-virulent ISAV. At early, peak, and late infection, mean ratios of spliced to unspliced segment 7 RNA were 3.0 % (± 0.7), 1.7 % (± 0.3), and 1.5 % (± 0.1) for the low virulent and 9.4 % (± 2.2), 4.7 % (± 0.8), and 6.2 % (± 0.1) for the high virulent isolate, respectively. By detection and quantification of active ISAV transcription, this assay may provide a more detailed understanding of ISAV infection dynamics.


Asunto(s)
Enfermedades de los Peces , Isavirus , Infecciones por Orthomyxoviridae , Salmo salar , Animales , Isavirus/genética , ARN Mensajero/genética , Infecciones por Orthomyxoviridae/diagnóstico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedades de los Peces/diagnóstico , Salmo salar/genética
15.
Front Immunol ; 14: 1158077, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37180109

RESUMEN

Many sialic acid-binding viruses express a receptor-destroying enzyme (RDE) that removes the virus-targeted receptor and limits viral interactions with the host cell surface. Despite a growing appreciation of how the viral RDE promotes viral fitness, little is known about its direct effects on the host. Infectious salmon anemia virus (ISAV) attaches to 4-O-acetylated sialic acids on Atlantic salmon epithelial, endothelial, and red blood cell surfaces. ISAV receptor binding and destruction are effectuated by the same molecule, the haemagglutinin esterase (HE). We recently discovered a global loss of vascular 4-O-acetylated sialic acids in ISAV-infected fish. The loss correlated with the expression of viral proteins, giving rise to the hypothesis that it was mediated by the HE. Here, we report that the ISAV receptor is also progressively lost from circulating erythrocytes in infected fish. Furthermore, salmon erythrocytes exposed to ISAV ex vivo lost their capacity to bind new ISAV particles. The loss of ISAV binding was not associated with receptor saturation. Moreover, upon loss of the ISAV receptor, erythrocyte surfaces became more available to the lectin wheat germ agglutinin, suggesting a potential to alter interactions with endogenous lectins of similar specificity. The pruning of erythrocyte surfaces was inhibited by an antibody that prevented ISAV attachment. Furthermore, recombinant HE, but not an esterase-silenced mutant, was sufficient to induce the observed surface modulation. This links the ISAV-induced erythrocyte modulation to the hydrolytic activity of the HE and shows that the observed effects are not mediated by endogenous esterases. Our findings are the first to directly link a viral RDE to extensive cell surface modulation in infected individuals. This raises the questions of whether other sialic acid-binding viruses that express RDEs affect host cells to a similar extent, and if such RDE-mediated cell surface modulation influences host biological functions with relevance to viral disease.


Asunto(s)
Isavirus , Salmo salar , Animales , Isavirus/fisiología , Ácidos Siálicos , Ácido N-Acetilneuramínico , Esterasas , Eritrocitos
16.
Vet Res ; 43: 27, 2012 Apr 09.
Artículo en Inglés | MEDLINE | ID: mdl-22486941

RESUMEN

Aquaculture is the fastest growing food production sector in the world. However, the increased production has been accompanied by the emergence of infectious diseases. Heart and skeletal muscle inflammation (HSMI) is one example of an emerging disease in farmed Atlantic salmon (Salmo salar L). Since the first recognition as a disease entity in 1999 it has become a widespread and economically important disease in Norway. The disease was recently found to be associated with infection with a novel reovirus, piscine reovirus (PRV). The load of PRV, examined by RT-qPCR, correlated with severity of HSMI in naturally and experimentally infected salmon. The disease is characterized by epi-, endo- and myocarditis, myocardial necrosis, myositis and necrosis of the red skeletal muscle. The aim of this study was to investigate the presence of PRV antigens in heart tissue of Atlantic salmon and monitor the virus distribution in the heart during the disease development. This included target cell specificity, viral load and tissue location during an HSMI outbreak. Rabbit polyclonal antisera were raised against putative PRV capsid proteins µ1C and σ1 and used in immunohistochemical analysis of archived salmon heart tissue from an experimental infection. The results are consistent with the histopathological changes of HSMI and showed a sequential staining pattern with PRV antigens initially present in leukocyte-like cells and subsequently in cardiomyocytes in the heart ventricle. Our results confirm the association between PRV and HSMI, and strengthen the hypothesis of PRV being the causative agent of HSMI. Immunohistochemical detection of PRV antigens will be beneficial for the understanding of the pathogenesis of HSMI as well as for diagnostic purposes.


Asunto(s)
Enfermedades de los Peces/virología , Miocarditis/veterinaria , Infecciones por Reoviridae/veterinaria , Reoviridae/genética , Salmo salar , Animales , Antígenos Virales/análisis , Proteínas de la Cápside/genética , Proteínas de la Cápside/metabolismo , Escherichia coli/genética , Corazón/virología , Miocarditis/virología , Conejos , Reoviridae/aislamiento & purificación , Reoviridae/metabolismo , Infecciones por Reoviridae/virología , Distribución Tisular , Carga Viral/veterinaria
17.
Fish Shellfish Immunol ; 32(1): 141-50, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22100613

RESUMEN

Development of diagnostic and prophylactic methodologies is dependent on knowledge of the host's defence system and reaction to different vaccine adjuvants. Here we present a sequential morphological study of peritonitis and inflammatory cell processing of incomplete Freund's adjuvant (IFA) in intraperitoneally injected Atlantic cod. The peritoneal tissue responses were characterised using necropsy, histology and electron microscopy. An extensive inflammatory response as characterised by leukocyte morphology and contents of enzymes, presence of apoptotic cells and IFN-γ-expressing cells was observed. Three days post injection, IFA droplets were surrounded by different types of inflammatory cells and two different patterns could be discerned. The first was characterised by flattened and concentrically arranged interdigitating cells connected by desmosomes and with macrophage-like cells (MLCs) predominant in the periphery. The second type possessed four stratified layers with an inner layer containing many apoptotic MLCs; a second layer containing flattened and shrunken cells and outer layers comprising moderately flattened cells and an outermost layer of mononuclear cells expressing IFN-γ. Oil was detected both inside and outside MLCs. The two types of processes, of which the second was clearly stratified, were similar to those observed in other teleosts, indicating a variety of reaction modes or alternatively sequential process development. The numerous dead MLCs contributed to inflammation.


Asunto(s)
Enfermedades de los Peces/inducido químicamente , Enfermedades de los Peces/patología , Adyuvante de Freund , Gadus morhua/inmunología , Lípidos , Peritonitis/veterinaria , Animales , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/mortalidad , Interferón gamma/inmunología , Microscopía Electrónica de Transmisión , Peritonitis/inducido químicamente , Peritonitis/inmunología , Peritonitis/mortalidad , Peritonitis/patología , ARN Mensajero/metabolismo , Temperatura
18.
Viruses ; 14(2)2022 02 02.
Artículo en Inglés | MEDLINE | ID: mdl-35215905

RESUMEN

Infectious salmon anaemia virus (ISAV) binds circulating Atlantic salmon erythrocytes, but the relevance of this interaction for the course of infection and development of disease remains unclear. We here characterise ISAV-erythrocyte interactions in experimentally infected Atlantic salmon and show that ISAV-binding to erythrocytes is common and precedes the development of disease. Viral RNA and infective particles were enriched in the cellular fraction of blood. While erythrocyte-associated ISAV remained infectious, erythrocytes dose-dependently limited the infection of cultured cells. Surprisingly, immunostaining of blood smears revealed expression of ISAV proteins in a small fraction of erythrocytes in one of the examined trials, confirming that ISAV can be internalised in this cell type and engage the cellular machinery in transcription and translation. However, viral protein expression in erythrocytes was rare and not required for development of disease and mortality. Furthermore, active transcription of ISAV mRNA was higher in tissues than in blood, supporting the assumption that ISAV replication predominantly takes place in endothelial cells. In conclusion, Atlantic salmon erythrocytes bind ISAV and sequester infective virus particles during infection, but do not appear to significantly contribute to ISAV replication. We discuss the implications of our findings for infection dynamics and pathogenesis of infectious salmon anaemia.


Asunto(s)
Eritrocitos/virología , Enfermedades de los Peces/virología , Isavirus/fisiología , Infecciones por Orthomyxoviridae/veterinaria , Salmo salar/virología , Animales , Células Endoteliales/virología , Enfermedades de los Peces/sangre , Isavirus/genética , Isavirus/aislamiento & purificación , Infecciones por Orthomyxoviridae/sangre , Infecciones por Orthomyxoviridae/virología , Salmo salar/sangre , Proteínas Virales/genética , Proteínas Virales/metabolismo , Virión/genética , Virión/aislamiento & purificación , Virión/fisiología , Replicación Viral
19.
J Gen Virol ; 92(Pt 4): 909-18, 2011 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-21148272

RESUMEN

Infectious salmon anemia virus (ISAV) is an orthomyxovirus responsible for a significant disease of farmed Atlantic salmon. Fallowing and re-establishment of the Atlantic salmon farming industry in the Faroes following a recent devastating infectious salmon anaemia (ISA) disease epidemic provided a unique opportunity to study the risk of re-emergence of disease. Over 53 months, 2787 of 34 573 (8.1%) apparently healthy Atlantic salmon analysed tested positive for ISAV by RT-PCR. Sequence analysis revealed the putative low-pathogenic ISAV-HPR0 subtype in all cases. Results demonstrated that ISAV-HPR0 appeared as a seasonal and transient infection without detectable ISA mortality or pathology. This finding, coupled to an apparent gill tropism of ISAV-HPR0, suggests ISAV-HPR0 causes a subclinical respiratory infection more like seasonal influenza, as opposed to the systemic infection and serious disease caused by highly pathogenic ISAV. The mean time before marine sites became infected was 7.7 months after transfer to seawater of the fish, suggesting a potentially unknown marine reservoir of infection. Sequence analysis identified two main subtypes of ISAV-HPR0 sequences, one of which showed close genetic association with ISAV isolates responsible for the disease outbreak in the Faroes. Thus ISAV-HPR0 might represent an ancestor of pathogenic variants and thus be a potential risk factor in the emergence of new strains of disease-causing ISAV. Our data, however, suggest that the risk of emergence of pathogenic ISAV variants from a reservoir of ISAV-HPR0 is low. This risk is probably being further reduced by practical management strategies adopted in the Faroes and aimed at reducing the potential for maintenance and adaptation of ISAV-HPR0.


Asunto(s)
Portador Sano/epidemiología , Portador Sano/virología , Isavirus/aislamiento & purificación , Isavirus/patogenicidad , Salmo salar/virología , Animales , Análisis por Conglomerados , Genotipo , Branquias/virología , Epidemiología Molecular , Datos de Secuencia Molecular , Prevalencia , ARN Viral/genética , Análisis de Secuencia de ADN
20.
Arch Virol ; 156(12): 2285-9, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21935625

RESUMEN

The putatively avirulent infectious salmon anaemia virus (ISAV) HPR0 variant has key phenotypic differences to isolates from disease outbreaks in Atlantic salmon farms. It appears to not cause disease, potentially displays a different tissue tropism and has yet to be isolated in conventional ISAV-permissive cell lines. This study focussed on identifying the biological basis for the observed differences by examining the properties of the haemagglutinin-esterase (HE) proteins derived from NWM10 (HPR0), Nevis 390/98 (HPR7 pathogenic strain) and mutant combinations of the two. Using a transfection-based system and haemadsorption analysis in salmon cell lines, this study demonstrated for the first time that an HPR0 HE was fully functional in terms of receptor-binding and -destroying activity and also suggested that the presence of a full-length HPR alone did not appear to affect these functions.


Asunto(s)
Enfermedades de los Peces/virología , Hemaglutininas Virales/genética , Isavirus/enzimología , Isavirus/genética , Infecciones por Orthomyxoviridae/veterinaria , Salmo salar/virología , Proteínas Virales de Fusión/genética , Animales , Línea Celular , Pruebas de Inhibición de la Hemadsorción , Técnicas In Vitro , Isavirus/patogenicidad , Infecciones por Orthomyxoviridae/virología , Polimorfismo Genético , Conejos , Receptores Virales/fisiología , Virulencia/genética
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