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1.
Gene ; 723: 144134, 2020 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-31589960

RESUMO

Viral kinases are known to undergo autophosphorylation and also phosphorylate viral and host substrates. Viral kinases have been implicated in various diseases and are also known to acquire host kinases for mimicking cellular functions and exhibit virulence. Although substantial analyses have been reported in the literature on diversity of viral kinases, there is a gap in the understanding of sequence and structural similarity among kinases from different classes of viruses. In this study, we performed a comprehensive analysis of protein kinases encoded in viral genomes. Homology search methods have been used to identify kinases from 104,282 viral genomic datasets. Serine/threonine and tyrosine kinases are identified only in 390 viral genomes. Out of seven viral classes that are based on nature of genetic material, only viruses having double-stranded DNA and single-stranded RNA retroviruses are found to encode kinases. The 716 identified protein kinases are classified into 63 subfamilies based on their sequence similarity within each cluster, and sequence signatures have been identified for each subfamily. 11 clusters are well represented with at least 10 members in each of these clusters. Kinases from dsDNA viruses, Phycodnaviridae which infect green algae and Herpesvirales that infect vertebrates including human, form a major group. From our analysis, it has been observed that the protein kinases in viruses belonging to same taxonomic lineages form discrete clusters and the kinases encoded in alphaherpesvirus form host-specific clusters. A comprehensive sequence and structure-based analysis enabled us to identify the conserved residues or motifs in kinase catalytic domain regions across all viral kinases. Conserved sequence regions that are specific to a particular viral kinase cluster and the kinases that show close similarity to eukaryotic kinases were identified by using sequence and three-dimensional structural regions of eukaryotic kinases as reference. The regions specific to each viral kinase cluster can be used as signatures in the future in classifying uncharacterized viral kinases. We note that kinases from giant viruses Marseilleviridae have close similarity to viral oncogenes in the functional regions and in putative substrate binding regions indicating their possible role in cancer.


Assuntos
Proteínas Quinases/química , Proteínas Quinases/genética , Vírus/classificação , Domínio Catalítico , Biologia Computacional/métodos , Bases de Dados de Proteínas , Variação Genética , Fosforilação , Filogenia , Proteínas Quinases/metabolismo , Homologia de Sequência de Aminoácidos , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/metabolismo , Fatores de Virulência/química , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Vírus/enzimologia , Vírus/patogenicidade
2.
Arch Virol ; 164(11): 2823-2828, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31485748

RESUMO

A 278-bp region upstream of the beet curly top virus-SpCT (BCTV-SpCT) C2/C3 genes is necessary for promoter activity and exhibits significant sequence similarity to AL2/3 promoter sequences in tomato golden mosaic virus (TGMV). Maximal expression of the downstream C2/3 genes in BCTV-SpCT requires the presence of the C1 protein, which is supported by observations that mutation of the initiator codon for C1 results in decreased C2/C3 expression. This is similar to TGMV and cabbage leaf curl virus, where AL1 is required for maximal AL2/3 expression. Together, these data suggest a common strategy for complementary-sense gene regulation amongst curtoviruses and begomoviruses.


Assuntos
Begomovirus/genética , Geminiviridae/genética , Regulação Viral da Expressão Gênica/genética , Begomovirus/metabolismo , Sítios de Ligação/genética , Geminiviridae/metabolismo , Regiões Promotoras Genéticas/genética , Proteínas Virais/genética
3.
J Agric Food Chem ; 67(41): 11380-11387, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31535865

RESUMO

Southern rice black-streaked dwarf virus (SRBSDV) causes disease in crops, which reduces the quality and yield. Several commercial antiviral agents are available to control the SRBSDV induced disease. However, the mechanism of antiviral agents controlling SRBSDV is largely unknown. Identifying targets in SRBSDV is a key step of antiviral agent discovery. Here, we investigated the potential protein target of the antiviral agent dufulin. We cloned and expressed a soluble viroplasmic P6 protein in the prokaryote Escherichia coli and the eukaryote Spodoptera frugiperda 9. The dissociation constants of dufulin with the purified P6 protein from E. coli and S. frugiperda 9 expression systems were 4.49 and 4.95 µM, respectively, indicating a strong binding affinity between dufulin and P6 protein. In vivo, dufulin significantly inhibited the expression of both P6 protein and P6 gene in the SRBSDV-infected rice leaves. This inhibition on P6 protein expression was also observed in transformed Nicotiana benthamiana where the P6 was overexpressed. Our data also showed that dufulin inhibited the duplication of SRBSDV in a dose-dependent manner in infected rice leaves with a half maximum effective concentration of 3.32 mM. It is therefore concluded that dufulin targets the viroplasmic protein P6 to inhibit the virulence of SRBSDV.


Assuntos
Antivirais/farmacologia , Benzotiazóis/farmacologia , Doenças das Plantas/virologia , Reoviridae/efeitos dos fármacos , Proteínas Virais/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Oryza/virologia , Reoviridae/genética , Reoviridae/metabolismo , Proteínas Virais/genética
4.
Arch Virol ; 164(12): 3107-3109, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31529221

RESUMO

The lumpy skin disease virus (LSDV) vaccine, Herbivac LS, batch 008, was sequenced and found to differ from the Neethling vaccine strain in the locus encoding a superoxide dismutase (SOD) homolog. The presence of a SOD homolog, be it full-length (as in Herbivac LS) or truncated (as in Neethling) may affect vaccine immunogenicity.


Assuntos
Vírus da Doença Nodular Cutânea/genética , Superóxido Dismutase/genética , Sequenciamento Completo do Genoma/métodos , Animais , Bovinos , Tamanho do Genoma , Vírus da Doença Nodular Cutânea/classificação , Vírus da Doença Nodular Cutânea/enzimologia , Mutação , Proteínas Virais/genética , Vacinas Virais/genética
5.
Acta Virol ; 63(3): 301-308, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31507196

RESUMO

Transmissible gastroenteritis virus (TGEV) causes great economic loss to swine industry worldwide. Vaccination is an important method to control the TGEV infection. In this study, a TGEV oral vaccine was generated by transferring a eukaryotic expression recombinant plasmid carrying the SAD (A and D antigenic sites of the S protein) epitope of TGEV into a swine-origin Lactobacillus acidophilus (L. acidophilus). In orally immunized BALB/c mice, the TGEV L. acidophilus oral vaccine induced significantly higher level of SIgA antibodies specific to TGEV compared with the mice immunized with a commercial inactivated TGEV vaccine and similar levels of IgG specific to TGEV as the inactivated vaccine. Furthermore, the TGEV L. acidophilus oral vaccine induced higher levels of IFN-γ, which suggested that the vaccine was able to induce immune response. In brief, this novel TGEV L. acidophilus oral vaccine could induce high levels of both mucosal and humoral immune responses, which has a potential to be used in the pig industries in the future. Keywords: transmissible gastroenteritis virus (TGEV); live L. acidophilus oral vaccine; SIgA antibody; IgG antibody; IFN-γ; IL-4.


Assuntos
Anticorpos Antivirais , Epitopos , Gastroenterite Suína Transmissível , Lactobacillus acidophilus , Vírus da Gastroenterite Transmissível , Vacinas Virais , Administração Oral , Animais , Anticorpos Antivirais/sangue , Epitopos/genética , Epitopos/imunologia , Gastroenterite Suína Transmissível/imunologia , Gastroenterite Suína Transmissível/patologia , Imunogenicidade da Vacina/imunologia , Lactobacillus acidophilus/genética , Lactobacillus acidophilus/virologia , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos/genética , Suínos , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/imunologia
6.
Arch Virol ; 164(10): 2637-2640, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31372754

RESUMO

A novel lytic Raoultella phage, RP180, was isolated and characterized. The RP180 genome has 44,851 base pairs and contains 65 putative genes, 35 of them encoding proteins whose functions were predicted based on sequence similarity to known proteins. The RP180 genome possesses a gene synteny typical of members of the subfamily Guernseyvirinae. Phylogenetic analysis of the RP180 genome and similar phage genomes revealed that phage RP180 is the first member of the genus Kagunavirus, subfamily Guernseyvirinae, that is specific for Raoultella sp. The genome of RP180 encodes a putative protein with similarity to CRISPR-like Cas4 nucleases, which belong to the pfam12705/PDDEXK_1 family. Cas4-like proteins of this family have been shown to interfere with the bacterial host type II-C CRISPR-Cas system.


Assuntos
Bacteriófagos/classificação , Bacteriófagos/isolamento & purificação , Enterobacteriaceae/virologia , Filogenia , Siphoviridae/classificação , Siphoviridae/isolamento & purificação , Bacteriólise , Bacteriófagos/genética , Genoma Viral , Microscopia Eletrônica de Transmissão , Análise de Sequência de DNA , Siphoviridae/genética , Sintenia , Proteínas Virais/genética , Vírion/ultraestrutura
7.
Arch Virol ; 164(10): 2641-2644, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31375908

RESUMO

We report the complete genome sequence of a novel nucleorhabdovirus, apple rootstock virus A (ApRVA), isolated from Malus spp. in South Korea. ApRVA has a 14,043-nt single-stranded negative-sense RNA genome. In the antigenome sense, it contains seven open reading frames, encoding the putative nucleocapsid protein, phosphoprotein, cell-to-cell movement protein, matrix protein, glycoprotein, RNA-dependent RNA polymerase, and an additional hypothetical protein, the gene for which is located between the genes for the matrix protein and glycoprotein. The complete genome sequence of ApRVA showed 47.45% nucleotide sequence identity to that of black currant-associated rhabdovirus 1. The genome organization, phylogenetic relationships, and sequence similarities to other nucleorhabdoviruses suggest that ApRVA is a new member of the genus Nucleorhabdovirus.


Assuntos
Genoma Viral , Malus/virologia , Raízes de Plantas/virologia , Rhabdoviridae/classificação , Rhabdoviridae/genética , Análise de Sequência de DNA , Ordem dos Genes , Fases de Leitura Aberta , Filogenia , RNA Viral/genética , República da Coreia , Rhabdoviridae/isolamento & purificação , Homologia de Sequência do Ácido Nucleico , Proteínas Virais/genética
8.
Arch Virol ; 164(10): 2585-2592, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31377889

RESUMO

Marbled eel reovirus (MERV) is an aquareovirus (AQRV) isolated from diseased marbled eels (Anguilla marmorata) with petechial skin hemorrhage. In this study, we propagated MERV in a cell line derived from the brain of Aequidens rivulatus and purified viral particles by using a discontinuous cesium chloride gradient. Genomic RNA sequences were obtained through next-generation sequencing. MERV, similar to most other AQRVs, showed the presence of 11 double-stranded RNA segments encoding 12 proteins; however, the genome sequence displayed very little similarity to known AQRV sequences. Furthermore, the structural proteins of MERV were most closely related to American grass carp reovirus with sequence identity values of no more than 64.89%. Phylogenetic analysis based on the sequences of structural proteins indicated that MERV shows an evolutionary history between AQRV-B and -G, which belong to the saline and freshwater environment subgroups, respectively. We also observed that MERV showed a closer relationship to orthoreoviruses based on the protein sequences of NS38 and NS73. In summary, MERV is a novel AQRV that could be classified as a member of the new proposed AQRV species "Aquareovirus H". The taxonomic assignments and evolution of AQRVs thus warrant further investigation.


Assuntos
Anguilla/virologia , Doenças dos Peixes/virologia , Infecções por Reoviridae/veterinária , Reoviridae/isolamento & purificação , Animais , Encéfalo/virologia , Linhagem Celular , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Microscopia Eletrônica de Transmissão , Filogenia , RNA de Cadeia Dupla/genética , RNA Viral/genética , Reoviridae/classificação , Reoviridae/genética , Infecções por Reoviridae/virologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Coloração e Rotulagem , Proteínas Virais/genética , Vírion/ultraestrutura , Cultura de Vírus
9.
Arch Virol ; 164(11): 2865-2871, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401694

RESUMO

Phage Vp_R1 belongs to the family Podoviridae and has a C3 morphotype, with an elongated head with a diameter of 190 ± 1.1 nm and an ultrashort tail with a length of 9 ± 1.2 nm. The double-stranded DNA genome is 112.1 kb long, has a mol% G + C content of 40.3, contains 129 ORFs, and encodes four tRNAs. Phylogenetic analysis suggests that phage Vp_R1 is a novel member of the genus Kuravirus.


Assuntos
Genoma Viral/genética , Podoviridae/genética , Vibrio parahaemolyticus/virologia , Sequência de Aminoácidos , Composição de Bases/genética , DNA Viral/genética , Fases de Leitura Aberta/genética , Podoviridae/isolamento & purificação , Análise de Sequência de DNA , Proteínas Virais/genética
10.
Arch Virol ; 164(11): 2881-2885, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31456087

RESUMO

Aquatic birds are known to be a reservoir for the most common influenza A viruses (IAVs). In the annual surveillance program, we collected the feces of migratory birds for the detection of IAVs in South Korea in November 2016. A novel reassorted H3N3 avian influenza virus (AIV) containing genes from viruses of wild and domestic birds was identified and named A/aquatic bird/South Korea/sw006/2016(H3N3). The polymerase basic 2 (PB2) and non-structural (NS) genes of this isolate are most closely related to those of wild-bird-origin AIV, while the polymerase basic 1 (PB1), polymerase acidic (PA), hemagglutinin (HA), nucleoprotein (NP), neuraminidase (NA), and matrix (M) genes are most closely related to those of domestic-bird-origin AIV. A/aquatic bird/South Korea/sw006/2016 contains PA, NP, M, and NS genes were most closely related to those of AIV subtype H4 and PB2, PB1, and HA genes that are most closely related to those of AIV subtype H3N8, while the NA gene was most closely related to those of subtype H10, which was recently detected in humans in China. These results suggest that novel reassortment of AIV strains occurred due to interaction between wild and domestic birds. Hence, we emphasize the need for continued surveillance of avian influenza virus in bird populations.


Assuntos
Genoma Viral/genética , Vírus da Influenza A Subtipo H3N8/genética , Influenza Aviária/virologia , Vírus Reordenados/genética , Animais , Aves/virologia , Vírus da Influenza A Subtipo H3N8/isolamento & purificação , Neuraminidase/genética , Proteínas não Estruturais Virais/genética , Proteínas Virais/genética , Sequenciamento Completo do Genoma
11.
Pan Afr Med J ; 32: 215, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31404285

RESUMO

Introduction: Kaposi's sarcoma (KS) is a kind of cancer that causes flat or raised lesions containing Human herpes virus 8 (HHV8). The KS lesions are common among immunosuppressed HIV patients. Highly Active Antiretroviral (HHART) treats and prevents the development of KS. The objective of this study was to determine the presence of K1 and K15 (predominant alleles) genes in Kaposi's sarcoma-associated herpes virus (KSHV) among immunosuppressed patients due to HIV-1. Methods: This was a cross-sectional descriptive study where consecutive sampling technique was adopted to pick archived tissue blocks from the Thematic Unit of Anatomic Pathology, Department of Human Pathology, College of Health Sciences, University of Nairobi and Department of Laboratory Medicine, Histology Section, Kenyatta National Hospital. Results: Upon staining 81 tissue blocks with H & E, 84% (68/81) were diagnosed as KS and 16% (13/81) as KS-like. The K1 and K15 (P) genes were both detected at 88.9% (72/81) in the tissue blocks, with 95.8% (69/72) detection from KS and 4.2% (3/72) from the KS-like. Conclusion: The K1 and K15 (P) genes of KSHV were present among the immunosuppressed patients with Human Immunodeficiency Virus (HIV)-1. It is important to carry out K1 and K15 (P) genes detection on tissues that are diagnosed as KS or KS-like by histology technique.


Assuntos
Infecções por HIV/complicações , Sarcoma de Kaposi/genética , Proteínas Virais/genética , Síndrome de Imunodeficiência Adquirida/complicações , Síndrome de Imunodeficiência Adquirida/tratamento farmacológico , Síndrome de Imunodeficiência Adquirida/genética , Adolescente , Adulto , Terapia Antirretroviral de Alta Atividade/métodos , Estudos Transversais , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/genética , HIV-1/isolamento & purificação , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Sarcoma de Kaposi/epidemiologia , Sarcoma de Kaposi/etiologia , Adulto Jovem
12.
J Microbiol Biotechnol ; 29(8): 1316-1323, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31434175

RESUMO

Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly emerging coronavirus which is zoonotic from bats and camels. Its infection in humans can be fatal especially in patients with preexisting conditions due to smoking and chronic obstructive pulmonary disease (COPD). Among the 25 proteins encoded by MERS-CoV, 5 accessory proteins seem to be involved in viral evasion of the host immune responses. Here we report that ORF4a, ORF4b, and ORF8b proteins, alone or in combination, effectively antagonize nuclear factor kappa B (NF-κB) activation. Interestingly, the inhibition of NF-κB by MERS-CoV accessory proteins was mostly at the level of pattern recognition receptors: melanoma differentiationassociated gene 5 (MDA5). ORF4a and ORF4b additively inhibit MDA5-mediated activation of NF-κB while that of retinoic acid-inducible gene 1 (RIG-I) is largely not perturbed. Of note, ORF8b was found to be a novel antagonist of MDA5-mediated NF-kκB activation. In addition, ORF8b also strongly inhibits Tank-binding kinase 1 (TBK1)-mediated induction of NF-κB signaling. Taken together, MERS-CoV accessory proteins are involved in viral escape of NF-κB-mediated antiviral immune responses.


Assuntos
Helicase IFIH1 Induzida por Interferon/metabolismo , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , NF-kappa B/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais , Animais , Infecções por Coronavirus/imunologia , Células HEK293 , Interações Hospedeiro-Patógeno , Humanos , Helicase IFIH1 Induzida por Interferon/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Virais/genética
13.
Dokl Biochem Biophys ; 486(1): 201-205, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31367821

RESUMO

Infection of mice with influenza A viruses led to the formation of clones of lymphocytes that specifically recognizes viral domains in the central zone of the NSP protein (amino acid positions 83-119). Computer analysis of the primary structure of the NSP protein showed the presence of T-cell epitopes in the central part of the NSP molecule. The findings indicate that the viral NSP gene is expressed in the infected animals and verify the concept of the bipolar strategy (ambisense strategy) of the influenza A virus genome.


Assuntos
Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/fisiologia , Vírus da Influenza A Subtipo H3N2/genética , Vírus da Influenza A Subtipo H3N2/fisiologia , Leucócitos/imunologia , RNA Viral/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Leucócitos/virologia , Camundongos , Camundongos Endogâmicos BALB C , Domínios Proteicos , Proteínas Virais/química , Proteínas Virais/metabolismo
14.
Virol J ; 16(1): 89, 2019 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-31277670

RESUMO

A tenuivirus, referred to here as JKI 29327, was isolated from a black medic (Medicago lupulina) plant collected in Austria. The virus was mechanically transmitted to Nicotiana benthamiana, M. lupulina, M. sativa, Pisum sativum and Vicia faba. The complete genome was determined by high throughput sequencing. The genome of JKI 29327 consists of eight RNA segments closely related to those of melon chlorotic spot virus (MeCSV) isolate E11-018 from France. Since segments RNA 7 and 8 of JKI 29327 are shorter, its genome is slightly smaller (by 247 nts) than that of E11-018. Pairwise comparisons between the predicted virus proteins of JKI 29327 and their homologues in E11-018 showed aa identities ranging from 80.6 to 97.2%. Plants infected with E11-081 gave intermediate DAS-ELISA reactions with polyclonal antibodies to JKI 29327. Since JKI 29327 and E11-018 appear to be closely related both serologically and genetically, we propose to regard JKI 29327 as the black medic strain of MeCSV. To our knowledge, JKI 29327 represents the second tenuivirus identified from a dicotyledonous plant. Serological and molecular diagnostic methods were developed for future detection.


Assuntos
Cucurbitaceae/virologia , Doenças das Plantas/virologia , Tenuivirus/genética , Tenuivirus/isolamento & purificação , Áustria , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Ervilhas/virologia , Filogenia , RNA Viral/genética , Tabaco/virologia , Vicia faba/virologia , Proteínas Virais/genética
15.
Virol J ; 16(1): 90, 2019 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-31319897

RESUMO

BACKGROUND: Nelson Bay orthoreovirus (NBV) was first isolated over 40 years ago from a fruit bat in Australia. Normally, NBV does not cause human diseases, but recently several NBV strains have been associated with human respiratory tract infections, thus attracting clinical attention. Autophagy, an evolutionarily conserved process in eukaryotic cells, degrades intracellular substrates, participates in multiple physiological processes, and maintains cellular homeostasis. In addition, autophagy is intimately involved in viral infection. METHODS: A new strain of NBV, isolated from a patient with a respiratory tract infection who returned to Japan from Bali, Indonesia, in 2007, was used in this study. NBV was rescued using a reverse genetics system involving cotransfection of BHK cells with 11 plasmids (pT7-L1 MB, pT7-L2 MB, pT7-L3 MB, pT7-M1 MB, pT7-M2 MB, pT7-M3 MB, pT7-S1 MB, pT7-S2 MB, pT7-S3 MB, pT7-S4 MB, and pcDNA3.1-T7), yielding NBV-MB. Recovered viruses were confirmed by immunofluorescence. The effect of NBV-MB on autophagy was evaluated by measuring the LC3-I/II proteins by immunoblot analysis after infection of BHK cells. Furthermore, after treatment with rapamycin (RAPA), 3-methyladenine (3-MA), chloroquine (CQ), or plasmid (GFP-LC3) transfection, the changes in expression of the LC3 gene and the amount of LC3-I/II protein were examined. In addition, variations in viral titer were assayed after treatment of BHK cells with drugs or after transfection with plasmids pCAGM3 and pCAGS3, which encode virus nonstructural proteins µNS and σNS, respectively. RESULTS: NBV-MB infection induced autophagy in host cells; however, the level of induction was dependent on viral replication. Induction of autophagy increased viral replication. By contrast, inhibiting autophagy suppressed NBV replication, albeit not significantly. The NBV-MB nonstructural protein µNS was involved in the induction of autophagy with viral infection. CONCLUSIONS: NBV-MB infection triggered autophagy. Also, the NBV nonstructural protein µNS may contribute to augmentation of autophagy upon viral infection.


Assuntos
Autofagia , Interações entre Hospedeiro e Microrganismos , Orthoreovirus/fisiologia , Replicação Viral , Linhagem Celular , Células HEK293 , Humanos , Infecções por Reoviridae/virologia , Genética Reversa , Carga Viral , Proteínas Virais/genética
16.
Vet Microbiol ; 235: 10-20, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282366

RESUMO

African Swine Fever Virus (ASFV) causes a hemorrhagic disease in swine and wild boars with a fatality rate close to 100%. Less virulent strains cause subchronic or chronic forms of the disease. The virus is endemic in sub-Saharan Africa and an outbreak in Georgia in 2007 spread to Armenia, Russia, Ukraine, Belarus, Poland, Lithuania, and Latvia. In August 2018, there was an outbreak in China and in April 2019, ASFV was reported in Vietnam and Cambodia. Since no vaccine or treatment exists, a vaccine is needed to safeguard the swine industry. Previously, we evaluated immunogenicity of two adenovirus-vectored cocktails containing ASFV antigens and demonstrated induction of unprecedented robust antibody and T cell responses, including cytotoxic T lymphocytes. In the present study, we evaluated protective efficacy of both cocktails by intranasal challenge of pigs with ASFV-Georgia 2007/1. A nine antigen cocktail-(I) formulated in BioMize adjuvant induced strong IgG responses, but when challenged, the vaccinees had more severe reaction relative to the controls. A seven antigen cocktail-(II) was evaluated using two adjuvants: BioMize and ZTS-01. The BioMize formulation induced stronger antibody responses, but 8/10 vaccinees and 4/5 controls succumbed to the disease or reached experimental endpoint at 17 days post-challenge. In contrast, the ZTS-01 formulation induced weaker antibody responses, but 4/9 pigs succumbed to the disease while the 5 survivors exhibited low clinical scores and no viremia at 17 days post-challenge, whereas 4/5 controls succumbed to the disease or reached experimental endpoint. Overall, none of the immunogens conferred statistically significant protection.


Assuntos
Febre Suína Africana/prevenção & controle , Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Vacinas Virais/imunologia , Adenoviridae , Administração Intranasal , Febre Suína Africana/imunologia , Vírus da Febre Suína Africana , Animais , Antígenos Virais/genética , Imunoglobulina G/sangue , Suínos , Linfócitos T Citotóxicos/imunologia , Vacinas de Subunidades/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/genética , Viremia , Virulência
17.
Vet Microbiol ; 235: 21-24, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282375

RESUMO

Occurrence of avian influenza (AI) with Neuraminidase (NA) mutations which confer reduced neuraminidase inhibitor (NAI) susceptibility has remained a cause of concern. The susceptibility to NAIs of 67 highly pathogenic avian influenza H5N1 viruses isolated during 2006-2012 in India was tested in phenotypic fluorescence-based NA inhibition assay, sequence analysis and in ovo. One isolate showed a novel NA I117T amino acid substitution (N2 numbering) and eight isolates showed previously known NAI-resistance marker mutations (I117V, E119D, N294S, total 9/67). The overall incidence of resistant variants was 13.4%. The novel I117T substitution reduced oseltamivir susceptibility by 18.6-fold and zanamivir susceptibility by 11.8-fold, compared to the wild type AI H5N1virus, thus showed cross-resistance to both oseltamivir and zanamivir in NA inhibition assays. However, the other two isolates with I117V substitution were sensitive to both the NAIs. In addition, the comparison of growth of the I117T and I117V variants in presence of NAI's in the in ovo assays exhibited difference in growth levels. The present study reports the natural occurrence of a novel I117T mutation in AI H5N1 virus conferring cross-resistance to oseltamivir and zanamivir highlighting the urgent need of antiviral surveillance of AI viruses.


Assuntos
Antivirais/farmacologia , Virus da Influenza A Subtipo H5N1/efeitos dos fármacos , Neuraminidase/genética , Oseltamivir/farmacologia , Proteínas Virais/genética , Zanamivir/farmacologia , Substituição de Aminoácidos , Animais , Galinhas , Farmacorresistência Viral , Índia , Virus da Influenza A Subtipo H5N1/genética , Concentração Inibidora 50 , Mutação de Sentido Incorreto , Zigoto
18.
Arch Virol ; 164(10): 2593-2597, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31270606

RESUMO

Trionyx sinensis hemorrhagic syndrome virus (TSHSV) is a newly discovered lethal arterivirus that causes serious disease in Trionyx sinensis in China. In this study, the complete genome sequence of TSHSV was determined by RACE cloning, and the functions of the predicted proteins were predicted. The complete genome of TSHSV was found to be 17,875 bp in length, and a 3'-end poly(A) tail was detected. Eight TSHSV hypothetical proteins (TSHSV-HPs) were predicted by gene model identification. TSHSV-HP2, 3 and 4 were associated with replicase activity, since papain-like protease (PLPs), serine-type endopeptidase, P-loop-containing nucleoside triphosphate hydrolase, and EndoU-like endoribonuclease motifs were detected. Phylogenetic analysis showed that TSHSV clusters with an arterivirus from a Chinese broad-headed pond turtle.


Assuntos
Infecções por Arterivirus/veterinária , Arterivirus/classificação , Arterivirus/isolamento & purificação , Filogenia , Tartarugas/virologia , Animais , Arterivirus/genética , Infecções por Arterivirus/virologia , China , Genoma Viral , RNA Mensageiro , Análise de Sequência de DNA , Proteínas Virais/genética
19.
Arch Virol ; 164(10): 2609-2611, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31312966

RESUMO

A new virus belonging to the genus Vitivirus in the family Betaflexiviridae was identified by next-generation sequencing of a blueberry plant showing green mosaic symptoms. The genome organization of the virus, which is tentatively named "blueberry green mosaic-associated virus" (BGMaV), is typical of vitiviruses, with five open reading frames (ORFs) and a polyadenylated 3' terminus. The ORFs code for the viral replicase, a 16K protein of unknown function, a movement protein, a coat protein (CP), and a nucleic acid binding protein. Phylogenetic analyses based on the deduced amino acid sequence of the CP and conserved motifs of the RNA-dependent RNA polymerase confirmed the taxonomic placement of BGMaV in the genus Vitivirus.


Assuntos
Mirtilos Azuis (Planta)/virologia , Flexiviridae/classificação , Flexiviridae/isolamento & purificação , Filogenia , Doenças das Plantas/virologia , Ordem dos Genes , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Anotação de Sequência Molecular , Fases de Leitura Aberta , RNA Mensageiro , RNA Viral/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Proteínas Virais/genética
20.
Arch Virol ; 164(10): 2617-2620, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31346768

RESUMO

The complete genome sequence of a trichovirus was obtained from peach samples collected from Mexico and found to be 7985 nucleotides long, excluding the poly(A) tail. Phylogenetic analysis using the complete nucleotide sequence revealed that the virus is a member of the genus Trichovirus and is closely related to peach mosaic virus (PcMV) and cherry mottle leaf virus (CMLV). The highest nucleotide sequence identity was 70% to both PcMV and CMLV, indicating that this virus, which we have tentatively named "peach virus M" (PeVM) should be considered a member of a new trichovirus species. We determined, for the first time, the initiation sites of the subgenomic RNAs (sgRNA) of a trichovirus. The sgRNAs for the movement and coat proteins started with the sequence 'GAA', while the smallest one, coding for the nucleotide-binding protein, started with the nucleotides 'GU'. In all cases, the sgRNAs leader ranged between 113 and 121 nt in length.


Assuntos
Flexiviridae/genética , Flexiviridae/isolamento & purificação , Filogenia , Prunus persica/virologia , Flexiviridae/classificação , México , RNA Viral/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Proteínas Virais/genética
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