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1.
Dev Comp Immunol ; 96: 135-143, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30885554

RESUMO

CD44 gene is a cell surface receptor which undergoes complex alternative splicing and extensive post-translational modifications. Although many studies have showed that CD44 is involved in the process of host defense, the function of piscine CD44 in antibacterial or antiviral defense response remains unclear. In the present study, we report the functional characterization of zebrafish CD44c, which is more similar to CD44b antigen isoforms rather than CD44a based on amino acid composition and phylogenetic analysis. The expression of zebrafish CD44c was inducible in response to bacterial and viral infections. During SVCV infection, the in vivo studies revealed that CD44c overexpression led to the increased virus loads and decreased survival rate. The attenuated response by zebrafish CD44c in response to SVCV infection were characterized by the impaired production of inflammatory cytokines and the impaired expressions of IFNs, IFN-stimulated genes, MHC class I and II genes. During Edwardsiella piscicida infection, the overexpression of zebrafish CD44c facilitated bacterial growth and dissemination, but did not impact on larvae survival. The detrimental role of CD44c in host defense against E. piscicida infection was supported by a decreased production of several antibacterial molecules including defbl2, defbl3, NK-lysin and RNase3. All together, these results firstly demonstrate the negative regulation of piscine CD44c in viral and bacterial infection.


Assuntos
Infecções por Enterobacteriaceae/imunologia , Doenças dos Peixes/imunologia , Receptores de Hialuronatos/imunologia , Infecções por Rhabdoviridae/imunologia , Peixe-Zebra/imunologia , Processamento Alternativo/imunologia , Sequência de Aminoácidos , Animais , Linhagem Celular , Citocinas/imunologia , Citocinas/metabolismo , Edwardsiella/imunologia , Edwardsiella/patogenicidade , Infecções por Enterobacteriaceae/microbiologia , Doenças dos Peixes/microbiologia , Receptores de Hialuronatos/química , Receptores de Hialuronatos/genética , Imunidade Inata , Larva/imunologia , Larva/metabolismo , Filogenia , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/imunologia , Rhabdoviridae/imunologia , Rhabdoviridae/patogenicidade , Infecções por Rhabdoviridae/virologia , Relação Estrutura-Atividade , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Peixe-Zebra/virologia
2.
Dev Comp Immunol ; 96: 68-77, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30853538

RESUMO

NOD-like receptor (NLR) family member X1 (NLRX1) of human localizes on mitochondria and serves as a negative regulator of antiviral signaling. However, the function of NLRX1 in teleost fish still remains elusive. To explore its role in the innate immunity of teleost fish, NLRX1 homologue has been cloned and characterized from black carp (Mylopharyngodon piceus). Black carp NLRX1 (bcNLRX1) consists of 1008 amino acids, which includes a N-terminal mitochondrial targeting sequence, a central NACHT domain and a C-terminal leucine-rich repeat (LRR) domain. bcNLRX1 was identified as a cytosolic protein locating on mitochondria through immunofluorescence (IF) staining. The overlapped subcellular distribution of bcNLRX1 and black carp MAVS (bcMAVS) was detected in IF staining, and the direct interaction between these two molecules in vitro was identified through co-immunoprecipitation assay. When co-expressed with bcMAVS, bcNLRX1 fiercely reduced bcMAVS-mediated IFN induction in reporter assay. Accordingly, the antiviral activity of bcMAVS against both grass carp reovirus (GCRV) and spring viremia of carp virus (SVCV) was forcefully repressed by bcNLRX1 in plaque assay. Mutagenic analyses further revealed that the NACHT domain of bcNLRX1 was essential for it to interact with bcMAVS and to suppress bcMAVS-mediated antiviral signaling. Taken together, our data support the conclusion that bcNLRX1 negatively regulates bcMAVS-mediated antiviral signaling through its NACHT domain during host innate immune activation.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Carpas/imunologia , Proteínas de Peixes/metabolismo , Proteínas Mitocondriais/metabolismo , Domínios Proteicos/imunologia , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Animais , Carpas/metabolismo , Carpas/virologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Proteínas de Peixes/imunologia , Células HEK293 , Humanos , Imunidade Inata , Proteínas Mitocondriais/imunologia , Ligação Proteica/imunologia , Reoviridae/imunologia , Reoviridae/patogenicidade , Rhabdoviridae/imunologia , Rhabdoviridae/patogenicidade , Transdução de Sinais/imunologia
3.
PLoS Pathog ; 15(3): e1007695, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30925159

RESUMO

p53, which regulates cell-cycle arrest and apoptosis, is a crucial target for viruses to release cells from cell-cycle checkpoints or to protect cells from apoptosis for their own benefit. Viral evasion mechanisms of aquatic viruses remain mysterious. Here, we report the spring viremia of carp virus (SVCV) degrading and stabilizing p53 in the ubiquitin-proteasome pathway by the N and P proteins, respectively. Early in an SVCV infection, significant induction was observed in the S phase and p53 was decreased in the protein level. Further experiments demonstrated that p53 interacted with SVCV N protein and was degraded by suppressing the K63-linked ubiquitination. However, the increase of p53 was observed late in the infection and experiments suggested that p53 was bound to SVCV P protein and stabilized by enhancing the K63-linked ubiquitination. Finally, lysine residue 358 was the key site for p53 K63-linked ubiquitination by the N and P proteins. Thus, our findings suggest that fish p53 is modulated by SVCV N and P protein in two distinct mechanisms, which uncovers the strategy for the subversion of p53-mediated host innate immune responses by aquatic viruses.


Assuntos
Rhabdoviridae/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Pontos de Checagem do Ciclo Celular/fisiologia , Vírus de DNA , Doenças dos Peixes , Regulação Viral da Expressão Gênica/genética , Células HEK293 , Humanos , Imunidade Inata , Rhabdoviridae/patogenicidade , Ubiquitinação , Viremia , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
4.
Adv Virus Res ; 102: 119-148, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30266171

RESUMO

A group of related bacilliform, nuclear viruses with a bisegmented negative-sense RNA genome that are transmitted by Brevipalpus mites likely in a circulative-propagative manner were recently classified in the new genus Dichorhavirus, family Rhabdoviridae. These viruses cause localized lesions on leaves, stems, and fruits of economically significant horticultural and ornamental plant species. Among its members, orchid fleck virus, citrus leprosis virus N, and coffee ringspot virus are most prominent. This chapter summarizes the current knowledge about these viruses, available detection techniques, and their interactions with their plant hosts and mite vectors.


Assuntos
Vetores Aracnídeos/virologia , Genoma Viral , Interações Hospedeiro-Patógeno , Ácaros/virologia , Plantas/virologia , Rhabdoviridae/genética , Animais , Mapeamento Cromossômico , Tipagem Molecular , Filogenia , Doenças das Plantas/virologia , RNA Viral/genética , RNA Viral/metabolismo , Rhabdoviridae/classificação , Rhabdoviridae/patogenicidade , Proteínas Virais/genética , Proteínas Virais/metabolismo , Replicação Viral
5.
Adv Virus Res ; 102: 23-57, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30266175

RESUMO

This chapter reviews the discoveries and initial characterizations (1930-1990) of three plant rhabdoviruses, sonchus yellow net virus, potato yellow dwarf virus, and lettuce necrotic yellows virus, that have become model systems for research on this group of enveloped negative-strand RNA plant viruses. We have used our personal perspectives to review the early historical studies of these viruses, the important technologies and tools, such as density gradient centrifugation, that were developed during the research, and to highlight the eminent scientists involved in these discoveries. Early studies on sites of virus replication, virion structure, physicochemical composition, and the use of protoplasts and vector insect cell culture for virus research are discussed, and differences between the nuclear and cytoplasmic lifestyles of plant rhabdoviruses are contrasted. Finally, we briefly summarize the genome organization and more recent developments culminating in the development of a reverse genetics system for plant negative-strand RNA viruses.


Assuntos
Genoma Viral , Doenças das Plantas/virologia , Patologia Vegetal/história , Rhabdoviridae/genética , Animais , Técnicas de Cultura de Células , História do Século XX , História do Século XXI , Insetos Vetores/citologia , Insetos Vetores/virologia , Modelos Biológicos , Vírus de Plantas/genética , Vírus de Plantas/metabolismo , Vírus de Plantas/patogenicidade , Vírus de Plantas/ultraestrutura , Plantas/virologia , Protoplastos/ultraestrutura , Protoplastos/virologia , Genética Reversa/métodos , Rhabdoviridae/metabolismo , Rhabdoviridae/patogenicidade , Rhabdoviridae/ultraestrutura , Vírion/genética , Vírion/metabolismo , Vírion/patogenicidade , Vírion/ultraestrutura
6.
Virus Res ; 255: 105-116, 2018 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-30009849

RESUMO

A batch of wild common carp and largemouth bass died in Andong, Gyeongsangbuk-do province, South Korea, in 2016. Moribund fish showed typical signs of spring viremia of carp (SVC) disease, which causes acute hemorrhage in the skin and ascites. Thus far, SVC disease has been detected in several regions of the world but never in South Korea. Suspecting the infectious agent to be the SCV virus (SVCV), the moribund fish were sampled and screened. The isolated virus developed a cytopathic effect in EPC cells. Both viral isolates from the common carp (ADC-SVC2016-1) and largemouth bass (ADC-SVC2016-3) were identical in terms of their genome sequence, which were 11,034 bp nucleotides in length. Genome comparison exhibited greater sequence similarity with the Asian SVCV sequences available at NCBI. Phylogenetic analysis revealed that the Korean SVCV isolates were clustered within the Asian clade. More specifically, evolutionary analysis by using the P gene sequences showed that the Korean isolates were sub-cladded within the Iai genogroup but diverged from Chinese strains of SH150514 and SH160901. The Korean isolates shared more than 98% sequence similarity with the two Chinese SVCV isolates, suggesting that the spread of SVCV originated from China. The isolated virus had cytopathic effects on EPC cells. Virus transmission studies showed that the virus exhibited the highest virulence at 15 °C, which was also dependent on the method used, with the injection method being better than the immersion and cohabitation methods. This is the first study to document that Korean SVCV isolates may be epizootic in wild common carp and other susceptible animal populations in South Korea.


Assuntos
Bass/virologia , Carpas/virologia , Doenças dos Peixes/virologia , Genoma , Filogenia , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae , Animais , Sequência de Bases , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/patologia , Genótipo , Dados de Sequência Molecular , República da Coreia/epidemiologia , Rhabdoviridae/classificação , Rhabdoviridae/genética , Rhabdoviridae/patogenicidade , Infecções por Rhabdoviridae/epidemiologia , Infecções por Rhabdoviridae/patologia , Infecções por Rhabdoviridae/virologia , Análise de Sequência de DNA , Virulência
7.
Mol Plant Pathol ; 19(10): 2288-2301, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29774653

RESUMO

Nucleorhabdoviruses such as Sonchus yellow net virus (SYNV) replicate in the nuclei and undergo morphogenesis at the inner nuclear membrane (IM) in plant cells. Mature particles are presumed to form by budding of the Matrix (M) protein-nucleocapsid complexes through host IMs to acquire host phospholipids and the surface glycoproteins (G). To address mechanisms underlying nucleorhabdovirus budding, we generated recombinant SYNV G mutants containing a truncated amino-terminal (NT) or carboxyl-terminal (CT) domain. Electron microscopy and sucrose gradient centrifugation analyses showed that the CT domain is essential for virion morphogenesis whereas the NT domain is also required for efficient budding. SYNV infection induces IM invaginations that are thought to provide membrane sites for virus budding. We found that in the context of viral infections, interactions of the M protein with the CT domain of the membrane-anchored G protein mediate M protein translocation and IM invagination. Interestingly, tethering the M protein to endomembranes, either by co-expression with a transmembrane G protein CT domain or by artificial fusion with the G protein membrane targeting sequence, induces IM invagination in uninfected cells. Further evidence to support functions of G-M interactions in virus budding came from dominant negative effects on SYNV-induced IM invagination and viral infections that were elicited by expression of a soluble version of the G protein CT domain. Based on these data, we propose that cooperative G-M interactions promote efficient SYNV budding.


Assuntos
Núcleo Celular/metabolismo , Glicoproteínas/metabolismo , Membrana Nuclear/metabolismo , Rhabdoviridae/patogenicidade
8.
Microbiol Immunol ; 61(10): 452-458, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28877365

RESUMO

The primordial immunoglobulin class, IgD, was the first non-IgM isotype discovered in teleosts. The crucial roles of IgM and IgZ in imparting systemic and mucosal immunity, respectively, in various fish species have been widely established. However, the putative function of a unique IgD isotype during pathogenic invasions has not been well explored. The present study reports the existence of an IgD ortholog in freshwater carp, Catla catla, and further evaluates its differential expression profile in response to bacterial, parasitic and viral antigenic exposure and pathogen associated molecular patterns (PAMPs) stimulation. The IgD of C. catla (CcIgD) cDNA sequence was found to encode 226 amino acids and confirmed homology with heavy chain delta region of Cyprinidae family members. Phylogenetic analysis of CcIgD exhibited greatest similarity with Ctenopharyngodon idella. qRT-PCR analysis revealed significant upregulation (P < 0.001) of IgD gene expression in kidney with respect to other tissues at 24 hr post-Aeromonas hydrophila challenge. CcIgD gene expression in skin was enhanced following Streptococcus uberis infection and in blood following Argulus infection and inactivated rhabdoviral antigen stimulation. Further, the treatment of bacterial and viral products (PAMPs) also triggered significant (P < 0.05) increases in CcIgD mRNA expression in kidney. These findings indicate the functional importance of teleost IgD in orchestrating tissue specific neutralization of antigens on stimulation with different pathogens and PAMPs.


Assuntos
Carpas/genética , Carpas/imunologia , Clonagem Molecular , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica , Imunoglobulina D/química , Imunoglobulina D/genética , Padrões Moleculares Associados a Patógenos , Aeromonas hydrophila/imunologia , Aeromonas hydrophila/patogenicidade , Sequência de Aminoácidos , Animais , Arguloida/patogenicidade , Infecções Bacterianas/imunologia , Cyprinidae/imunologia , DNA Complementar/genética , Doenças dos Peixes/microbiologia , Doenças dos Peixes/parasitologia , Doenças dos Peixes/virologia , Água Doce , Expressão Gênica , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Interações Hospedeiro-Parasita/genética , Interações Hospedeiro-Parasita/imunologia , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Imunoglobulina D/classificação , Cadeias Pesadas de Imunoglobulinas/química , Cadeias Pesadas de Imunoglobulinas/genética , Isotipos de Imunoglobulinas/química , Isotipos de Imunoglobulinas/genética , Rim , Doenças Parasitárias/imunologia , Filogenia , Rhabdoviridae/patogenicidade , Análise de Sequência de Proteína , Pele/imunologia , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/veterinária , Streptococcus/patogenicidade , Viroses/imunologia
9.
Virus Res ; 238: 183-192, 2017 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-28684157

RESUMO

Spring viraemia of carp virus (SVCV) is an OIE-listed notifiable pathogen, which has brought huge economic loss to the aquaculture industry. Outbreaks of SVC mostly occur in spring with water temperature 11-17°C. Presently, there is an increase in detection during import quarantine testing and associated with outbreaks of SVCV outside of China, yet China is regarded as the origin of SVCV Asian clade. However, recent isolates from the Shanghai area all showed to be low pathogenic to their original hosts. In this study, we isolated a new SVCV strain (nominated as SH160901) from grass carp in late summer in Shanghai, 2016. Phylogenetic analysis showed this strain formed a distinct new lineage in the Asian clade along with our isolate SH150514 in 2015, and was divergent from all other identified Asian isolates. Cell infection test demonstrated that this strain replicated most efficiently at 25°C and 28°C, and could induce obvious cytopathic effect in infected cells. In vivo infection test revealed this strain could cause severe symptoms in experimentally infected fish at 16-20°C. Inoculated fish died at 100% in grass carp and common carp (Cyprinus carpio carpio) within 13days, and at 100% in goldfish (Carassius auratus) and 90% in koi (Cyprinus carpio koi) within 40days. Experimental infections at 26°C also induced moderate mortalities in grass carp (25%) and common carp (20%). The biological changes characterized for SVCV isolate SH160901 warrant changes to surveillance plans, specifically there is a need to broaden the testing parameters previously associated with SVCV.


Assuntos
Carpas/virologia , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/isolamento & purificação , Rhabdoviridae/patogenicidade , Animais , China , Efeito Citopatogênico Viral , Genótipo , Filogenia , Rhabdoviridae/classificação , Rhabdoviridae/genética , Infecções por Rhabdoviridae/patologia , Infecções por Rhabdoviridae/virologia , Estações do Ano , Análise de Sobrevida , Temperatura , Cultura de Vírus , Replicação Viral
10.
Arch Virol ; 162(9): 2829-2834, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28550433

RESUMO

The Rhabdoviridae is a diverse family of negative-sense single-stranded RNA viruses which infects mammals, birds, reptiles, fish, insects and plants. Herein, we reported the isolation and characterization of 6 novel viruses from diseased fish collected from China including SCRV-QY, SCRV-SS, SCRV-GM, CmRV-FS, MsRV-SS, OmbRV-JM. The typical clinical symptom of diseased fish was hemorrhaging. Efficient propagation of these isolates in a Chinese perch brain cell line was determined by means of observation of cytopathic effect, RT-PCR and electron microscopy. Sequence alignment and phylogenetic analysis of the complete G protein sequences revealed that these isolates were clustered into one monophyletic lineage belonging to the species Siniperca chuatsi rhabdovirus.


Assuntos
Doenças dos Peixes/virologia , Variação Genética , Rhabdoviridae/genética , Rhabdoviridae/patogenicidade , Animais , China/epidemiologia , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/patologia , Peixes
11.
Eur J Neurosci ; 42(4): 2036-50, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25980955

RESUMO

Many RNA virus CNS infections cause neurological disease. Because Piry virus has a limited human pathogenicity and exercise reduces activation of microglia in aged mice, possible influences of environment and aging on microglial morphology and behavior in mice sublethal encephalitis were investigated. Female albino Swiss mice were raised either in standard (S) or in enriched (EE) cages from age 2 to 6 months (young - Y), or from 2 to 16 months (aged - A). After behavioral tests, mice nostrils were instilled with Piry-virus-infected or with normal brain homogenates. Brain sections were immunolabeled for virus antigens or microglia at 8 days post-infection (dpi), when behavioral changes became apparent, and at 20 and 40 dpi, after additional behavioral testing. Young infected mice from standard (SYPy) and enriched (EYPy) groups showed similar transient impairment in burrowing activity and olfactory discrimination, whereas aged infected mice from both environments (EAPy, SAPy) showed permanent reduction in both tasks. The beneficial effects of an enriched environment were smaller in aged than in young mice. Six-hundred and forty microglial cells, 80 from each group were reconstructed. An unbiased, stereological sampling approach and multivariate statistical analysis were used to search for microglial morphological families. This procedure allowed distinguishing between microglial morphology of infected and control subjects. More severe virus-associated microglial changes were observed in young than in aged mice, and EYPy seem to recover microglial homeostatic morphology earlier than SYPy . Because Piry-virus encephalitis outcomes were more severe in aged mice, it is suggested that the reduced inflammatory response in those individuals may aggravate encephalitis outcomes.


Assuntos
Envelhecimento , Encéfalo/patologia , Encefalite Viral/patologia , Encefalite Viral/terapia , Meio Ambiente , Microglia/patologia , Análise de Variância , Animais , Complexo CD3/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Modelos Animais de Doenças , Encefalite Viral/fisiopatologia , Comportamento Exploratório , Feminino , Imageamento Tridimensional , Memória/fisiologia , Camundongos , Proteínas dos Microfilamentos/metabolismo , Rhabdoviridae/patogenicidade , Olfato/fisiologia , Fatores de Tempo
12.
Trends Microbiol ; 23(5): 283-8, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25639651

RESUMO

Neuroinvasive viral infections invade the nervous system, often eliciting serious disease and death. Members of four viral families are both neuroinvasive and capable of transmitting progeny virions or virion components within the long neuronal extensions known as axons. Axons provide physical structures that enable viral infection to spread within the host while avoiding extracellular immune responses. Technological advances in the analysis of in vivo neural circuits, neuronal culturing, and live imaging of fluorescent fusion proteins have enabled an unprecedented view into the steps of virion assembly, transport, and egress involved in axonal spread. In this review we summarize the literature supporting anterograde (axon to cell) spread of viral infection, describe the various strategies of virion transport, and discuss the effects of spread on populations of neuroinvasive viruses.


Assuntos
Axônios/virologia , Neurônios/virologia , Vírus/patogenicidade , Animais , Flavivirus/patogenicidade , Genoma Viral , Herpesvirus Humano 1/patogenicidade , Humanos , Picornaviridae/patogenicidade , Rhabdoviridae/patogenicidade , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/fisiologia , Vírion/fisiologia , Montagem de Vírus
13.
Mikrobiol Z ; 76(2): 41-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25000729

RESUMO

Long-term studies of spread of rhabdoviruses which indicated their harmfulness in different plant species under conditions of environmental factors were first discussed. Their harmfulness to different plant species under environmental conditions was shown. A comparative description of rhabdoviruses with similar pathogens of the mushrooms is carried out. Thus the main focus was on the morphology and structure of the pathogens. These data are extremely important in the study of distribution of the rhabdovirus on crops in different regions.


Assuntos
Agaricales/virologia , Grão Comestível/virologia , Doenças das Plantas/virologia , Rhabdoviridae/ultraestrutura , Animais , Carpas/virologia , Doenças dos Peixes/virologia , Especificidade de Hospedeiro , Rhabdoviridae/patogenicidade , Especificidade da Espécie
14.
Ukr Biochem J ; 86(3): 33-40, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25033552

RESUMO

Ribonucleases (RNases) are present in base-level amounts in intact plants, but this level is able to increase greatly under stress conditions. The possible cause for such an increase is protection against plant RNA-virus attack. Buckwheat burn virus (BBV) is a highly virulent pathogen that belongs to Rhabdoviridae family. In our study, we have analyzed the correlation between RNase activity and resistance of different buckwheat cultivars to BBV infection. Two cultivars, Kara-Dag and Roksolana, with different sensitivities to BBV have been used. Kara-Dag is a cultivar with medium sensitivity to virus and Roksolana is a tolerant cultivar. It has been shown that the base level of RNase activity in Roksolana cultivar was in most cases higher than the corresponding parameter in Kara-Dag cultivar. Both infected and uninfected plants of Roksolana cultivar demonstrated high RNase activity during two weeks. Whereas infected plants of Kara-Dag cultivar demonstrated unstable levels of RNase activity. Significant decline in RNase activity was detected on the 7th day post infection with subsequent gradual increase in RNase activity. Decline of the RNase activity during the first week could promote the virus replication and therefore more successful infection of upper leaves of plants. Unstable levels of RNase activity in infected buckwheat plants may be explained by insufficiency of virus-resistant mechanisms that determines the medium sensitivity of the cultivar to BBV. Thus, plants of buckwheat cultivar having less sensitivity to virus, displayed in general higher RNase activity.


Assuntos
Fagopyrum/imunologia , Folhas de Planta/imunologia , Proteínas de Plantas/metabolismo , Ribonucleases/metabolismo , Fagopyrum/enzimologia , Fagopyrum/virologia , Interações Hospedeiro-Patógeno , Doenças das Plantas , Imunidade Vegetal , Folhas de Planta/enzimologia , Folhas de Planta/virologia , Proteínas de Plantas/imunologia , Rhabdoviridae/patogenicidade , Rhabdoviridae/fisiologia , Ribonucleases/imunologia
15.
Virus Res ; 189: 271-9, 2014 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-24954789

RESUMO

A new strain of spring viraemia of carp virus, denominated SVCV-265, was isolated from an ornamental common carp (Cyprinus carpio) in Shanghai, China, 2013. The isolate could produce obvious cytopathic effects on EPC cells, while was shown to be of low virulence for juvenile koi. Complete genome sequencing revealed the genome of the SVCV-265 strain is 11,029 nucleotides in length and phylogenetic analysis showed the isolate was clustered within Asia clade but was divergent from Chinese A1, A2 and BJ0505-2 strains. Previous report indicated that the G and P gene of SVCV shared similar topologies of evolutionary trees. In this study, phylogenetic analysis based on the P gene sequences showed the SVCV-265 was clustered into Iai subgroup and divergent from Chinese isolates A1, A2 and BJ0505-2, which were clustered into Iaii group. However, sequence alignment of the G gene showed the SVCV-265 has a close relationship with A1, A2 and BJ0505-2 isolates. Recombination analysis of all the whole sequences of SVCV available revealed isolates A2 and BJ0505-2 were likely the homologous recombination descendants of the A1 and SVCV-265. The crossover regions were located between 3845-6387nt for A2 and 3573-6444 nt for BJ0505-2, respectively. Phylogenetic analysis of the crossover region further confirmed these findings. This current study describes the molecular characterization of the new isolate SVCV-265 from China and is the first report of homologous recombination in SVCV.


Assuntos
Genoma Viral , Recombinação Homóloga , RNA Viral/genética , Rhabdoviridae/genética , Análise de Sequência de DNA , Animais , Carpas , Linhagem Celular , China , Análise por Conglomerados , Efeito Citopatogênico Viral , Dados de Sequência Molecular , Filogenia , Rhabdoviridae/isolamento & purificação , Rhabdoviridae/patogenicidade , Rhabdoviridae/fisiologia , Homologia de Sequência , Virulência
16.
Virus Res ; 189: 56-62, 2014 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-24794839

RESUMO

An Ussuri catfish Pseudobagrus ussuriensis skin (UCS) cell line was developed and subcultured for more than 60 passages. UCS cells consisted of mostly epithelial-like cells and multiplied well in TC199 medium supplemented with 10% fetal bovine serum at 25°C. Chromosome analysis revealed that most UCS cells had a normal diploid karyotype with 2n=52. UCS cells showed differential cytopathic effects (CPEs) after inoculation of spring viremia of carp virus (SVCV, a negative-strand RNA virus), grass carp reovirus (GCRV, a multi-segmented double-stranded RNA virus) and Rana grylio virus (RGV, a large double-stranded DNA virus), and were indicative of high sensitivities to these three aquatic animal viruses by a virus titration study. The CPE caused by SVCV appeared as rounded and granular cells, grape-like clusters and small lytic plaques. Characteristic CPE containing plaque-like syncytia was induced by GCRV. RGV-infected cells produced typical CPE characterized by cells shrinkage and aggregation, formation of clear plaques and cell sheet detachment. Furthermore, significant fluorescent signals were observed after UCS cells were transfected with green fluorescent protein reporter plasmids, and the development of CPE induced by a recombinant RGV, ΔTK-RGV, in UCS cells was illustrated using a combination of light and fluorescence microscopy. The data from this study suggested that UCS cell line can potentially serve as a useful tool for the comparison study of different aquatic animal viruses and the isolation of some newly emerging viruses in Ussuri catfish farming.


Assuntos
Peixes-Gato , Linhagem Celular , Efeito Citopatogênico Viral , Ranavirus/crescimento & desenvolvimento , Reoviridae/crescimento & desenvolvimento , Rhabdoviridae/crescimento & desenvolvimento , Pele , Animais , Meios de Cultura/química , Células Epiteliais/fisiologia , Células Epiteliais/virologia , Ranavirus/patogenicidade , Reoviridae/patogenicidade , Rhabdoviridae/patogenicidade , Temperatura , Ensaio de Placa Viral
18.
PLoS Genet ; 8(11): e1003057, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23166512

RESUMO

Variation in susceptibility to infectious disease often has a substantial genetic component in animal and plant populations. We have used genome-wide association studies (GWAS) in Drosophila melanogaster to identify the genetic basis of variation in susceptibility to viral infection. We found that there is substantially more genetic variation in susceptibility to two viruses that naturally infect D. melanogaster (DCV and DMelSV) than to two viruses isolated from other insects (FHV and DAffSV). Furthermore, this increased variation is caused by a small number of common polymorphisms that have a major effect on resistance and can individually explain up to 47% of the heritability in disease susceptibility. For two of these polymorphisms, it has previously been shown that they have been driven to a high frequency by natural selection. An advantage of GWAS in Drosophila is that the results can be confirmed experimentally. We verified that a gene called pastrel--which was previously not known to have an antiviral function--is associated with DCV-resistance by knocking down its expression by RNAi. Our data suggest that selection for resistance to infectious disease can increase genetic variation by increasing the frequency of major-effect alleles, and this has resulted in a simple genetic basis to variation in virus resistance.


Assuntos
Evolução Biológica , Resistência à Doença/genética , Drosophila melanogaster , Estudo de Associação Genômica Ampla , Alelos , Animais , Mapeamento Cromossômico , Dicistroviridae/genética , Dicistroviridae/patogenicidade , Drosophila melanogaster/genética , Drosophila melanogaster/virologia , Genótipo , Rhabdoviridae/genética , Rhabdoviridae/patogenicidade , Seleção Genética
19.
Vet Microbiol ; 157(3-4): 264-75, 2012 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-22243898

RESUMO

Plasmid DNAs containing Siniperca chuatsi rhabdovirus (SCRV) glycoprotein gene (pcDNA-G) and nucleoprotein gene (pcDNA-N) were constructed, and used to determine the antiviral immune response elicited by DNA vaccination in mandarin fish. In vitro and in vivo expression of the plasmid constructs was confirmed in transfected cells and muscle tissues of vaccinated fish by Western blot, indirect immunofluorescence or RT-PCR analysis. Fish injected with pcDNA-G exhibited protective effect against SCRV challenge with a relative percent survival (RPS) of 77.5%, but no significant protection (RPS of 2.5%) was observed in fish vaccinated with pcDNA-N. Immunohistochemical analysis showed that vaccination with pcDNA-G decreased histological lesions and suppressed the virus replication in fish target organs, e.g. kidney, liver, spleen, gill and heart. Transcriptional analysis further revealed that the expression levels of type I IFN system genes including interferon regulation factor-7 (IRF-7) gene, myxovirus resistance (Mx) gene and virus inhibitory protein (Viperin) gene were strongly up-regulated after injection with pcDNA-G, whereas the level of transcription of immunoglobulin M (IgM) gene did not show a statistically significant change. These results reveal that type I IFN antiviral immune response is rapidly triggered by the plasmid DNA containing rhabdovirus glycoprotein gene in fish, which offers an explanation of molecular mechanisms for DNA vaccination inducing mandarin fish resist to SCRV disease.


Assuntos
Doenças dos Peixes/prevenção & controle , Perciformes/imunologia , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/patogenicidade , Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Doenças dos Peixes/imunologia , Proteínas de Ligação ao GTP/imunologia , Glicoproteínas/genética , Glicoproteínas/imunologia , Imunoglobulina M/imunologia , Fator Regulador 7 de Interferon/imunologia , Interferon Tipo I/imunologia , Proteínas de Resistência a Myxovirus , Perciformes/virologia , Plasmídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/prevenção & controle , Vacinação/veterinária , Vacinas de DNA/genética , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/genética
20.
Uirusu ; 62(2): 183-96, 2012.
Artigo em Japonês | MEDLINE | ID: mdl-24153229

RESUMO

The family Rhabdoviridae has a non-segmented single stranded negative-sense RNA and its genome ranges in size from approximately 11 kb to almost 16 kb. It is one of the most ecologically diverse families of RNA viruses with members infecting a wide range of organisms. The five structural protein genes are arranged in the same linear order (3'-N-P-M-G-L-5') and may be interspersed with one more additional accessory gene. For many years, a full of knowledge of the rhabdoviridae has been established on extensive studies of two kinds of prototype viruses; vesicular stomatitis virus (VSV) and rabies virus (RABV). Among them, the genus Lyssavirus includes RABV and rabies-related viruses naturally infect mammals and chiropterans via bite-exposure by rabid animals and finally cause fatal encephalitis. In this review, we describe the sketch of the various virological features of the Rhabdoviridae, especially focusing on VSV and RABV.


Assuntos
Rhabdoviridae , Animais , Evolução Molecular , Proteínas de Ligação ao GTP , Genoma Viral/genética , Humanos , Raiva/virologia , Rhabdoviridae/classificação , Rhabdoviridae/genética , Rhabdoviridae/patogenicidade , Rhabdoviridae/fisiologia , Estomatite Vesicular/virologia , Proteínas Virais/fisiologia , Vírion , Replicação Viral/genética
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