Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.658
Filtrar
1.
Arch Virol ; 165(10): 2291-2299, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32754877

RESUMO

The multimammate mouse (Mastomys natalensis; M. natalensis) serves as the main reservoir for the zoonotic arenavirus Lassa virus (LASV), and this has led to considerable investigation into the distribution of LASV and other related arenaviruses in this host species. In contrast to the situation with arenaviruses, the presence of other viruses in M. natalensis remains largely unexplored. In this study, herpesviruses and polyomaviruses were identified and partially characterized by PCR methods, sequencing, and phylogenetic analysis. In tissues sampled from M. natalensis populations in Côte d'Ivoire and Mali, six new DNA viruses (four betaherpesviruses, one gammaherpesvirus and one polyomavirus) were identified. Phylogenetic analysis based on glycoprotein B amino acid sequences showed that the herpesviruses clustered with cytomegaloviruses and rhadinoviruses of multiple rodent species. The complete circular genome of the newly identified polyomavirus was amplified by PCR. Amino acid sequence analysis of the large T antigen or VP1 showed that this virus clustered with a known polyomavirus from a house mouse (species Mus musculus polyomavirus 1). These two polyomaviruses form a clade with other rodent polyomaviruses, and the newly identified virus represents the third known polyomavirus of M. natalensis. This study represents the first identification of herpesviruses and the discovery of a novel polyomavirus in M. natalensis. In contrast to arenaviruses, we anticipate that these newly identified viruses represent a low zoonotic risk due to the normally highly restricted specificity of members of these two DNA virus families to their individual mammalian host species.


Assuntos
Genoma Viral , Infecções por Herpesviridae/epidemiologia , Herpesviridae/genética , Filogenia , Infecções por Polyomavirus/epidemiologia , Polyomavirus/genética , Doenças dos Roedores/epidemiologia , África ao Sul do Saara/epidemiologia , Animais , Antígenos Virais de Tumores/genética , Proteínas do Capsídeo/genética , Reservatórios de Doenças/virologia , Herpesviridae/classificação , Herpesviridae/isolamento & purificação , Infecções por Herpesviridae/virologia , Especificidade de Hospedeiro , Tipagem Molecular , Murinae/virologia , Polyomavirus/classificação , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/virologia , Doenças dos Roedores/virologia , Proteínas do Envelope Viral/genética
2.
BMC Infect Dis ; 20(1): 488, 2020 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-32646445

RESUMO

BACKGROUND: Washington University polyomavirus (WUPyV) is a novel human polyomavirus detected in childwith acute respiratory infection in 2007. However, the relationship between WUPyV and respiratory diseases has yet to be established for lacking of a suitable in vitro culture system. METHODS: To isolate WUPyV with human airway epithelial (HAE) cells, the positive samples were incubated in HAE, and then the nucleic acid, VP1 protein and virions were detected using real-time PCR, immunofluorescence and electron microscopy respectively. RESULTS: The result showed that WUPyV could replicate effectively in HAE cells and virions with typical polyomavirus characteristics could be observed. Additionally, the entire genome sequence of the isolated strain (BJ0771) was obtained and phylogenetic analysis indicated that BJ0771 belongs to gene cluster I. CONCLUSIONS: Our findings demonstrated clinical WUPyV strain was successfully isolated for the first time in the world and this will help unravel the etiology and pathogenic mechanisms of WUPyV in respiratory infection diseases.


Assuntos
Células Epiteliais/virologia , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/virologia , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Mucosa Respiratória/patologia , Infecções Respiratórias/diagnóstico , Adolescente , Proteínas do Capsídeo/genética , Polaridade Celular , Células Cultivadas , Criança , Pré-Escolar , Células Epiteliais/metabolismo , Feminino , Humanos , Masculino , Família Multigênica , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Infecções Respiratórias/virologia , Vírion/genética , Replicação Viral , Sequenciamento Completo do Genoma
3.
J Virol ; 94(14)2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32404521

RESUMO

UNC5B is a dependence receptor that promotes survival in the presence of its ligand, netrin-1, while inducing cell death in its absence. The receptor has an important role in the development of the nervous and vascular systems. It is also involved in the normal turnover of intestinal epithelium. Netrin-1 and UNC5B are deregulated in multiple cancers, including colorectal, neuroblastoma, and breast tumors. However, the detailed mechanism of UNC5B function is not fully understood. We have utilized the murine polyomavirus small T antigen (PyST) as a tool to study UNC5B-mediated apoptosis. PyST is known to induce mitotic arrest followed by extensive cell death in mammalian cells. Our results show that the expression of PyST increases mRNA levels of UNC5B by approximately 3-fold in osteosarcoma cells (U2OS) and also stabilizes UNC5B at the posttranslational level. Furthermore, UNC5B is upregulated predominantly in those cells that undergo mitotic arrest upon PyST expression. Interestingly, although its expression was previously reported to be regulated by p53, our data show that the increase in UNC5B levels by PyST is p53 independent. The posttranslational stabilization of UNC5B by PyST is regulated by the interaction of PyST with PP2A. We also show that netrin-1 expression, which is known to inhibit UNC5B apoptotic activity, promotes survival of PyST-expressing cells. Our results thus suggest an important role of UNC5B in small-T antigen-induced mitotic catastrophe that also requires PP2A.IMPORTANCE UNC5B, PP2A, and netrin-1 are deregulated in a variety of cancers. UNC5B and PP2A are regarded as tumor suppressors, as they promote apoptosis and are deleted or mutated in many cancers. In contrast, netrin-1 promotes survival by inhibiting dependence receptors, including UNC5B, and is upregulated in many cancers. Here, we show that UNC5B-mediated apoptosis can occur independently of p53 but in a PP2A-dependent manner. A substantial percentage of cancers arise due to p53 mutations and are insensitive to chemotherapeutic treatments that activate p53. Unexpectedly, treatment of cancers having functional p53 with many conventional drugs leads to the upregulation of netrin-1 through activated p53, which is counterintuitive. Therefore, understanding the p53-independent mechanisms of the netrin-UNC5B axis, such as those involving PP2A, assumes greater clinical significance. Anticancer strategies utilizing anti-netrin-1 antibody treatment are already in clinical trials.


Assuntos
Antígenos Virais de Tumores/metabolismo , Apoptose , Receptores de Netrina/metabolismo , Polyomavirus/metabolismo , Proteína Fosfatase 2/metabolismo , Células A549 , Animais , Antígenos Virais de Tumores/genética , Células HeLa , Humanos , Camundongos , Receptores de Netrina/genética , Polyomavirus/genética , Proteína Fosfatase 2/genética
4.
PLoS Pathog ; 16(4): e1008523, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32324824

RESUMO

JC polyomavirus (JCPyV, JCV) causes progressive multifocal leukoencephalopathy (PML) in immunocompromised hosts. JCPyV replicates in oligodendrocytes within the brain tissue of patients with PML. The JCPyV genome encodes a microRNA (miRNA) in the region encoding the large T antigen. JCPyV-encoded miRNA (miR-J1) has been detected in the tissue and cerebrospinal fluid samples of patients with PML; however, there are no reports describing the localization of polyomavirus-encoded miRNA in histological samples of patients with virus-associated diseases. In the present study, we detected high miR-J1 expression in the nuclei of JCPyV-infected cells in PML tissue samples via in situ hybridization. Additionally, in situ hybridization also revealed the expression of BK polyomavirus (BKPyV, BKV)-encoded miRNA in lesions of BKPyV-associated nephropathy. In situ hybridization for miR-J1-5p and -3p showed positive signals in 24/25 (96%) of PML tissues that were positive for JCPyV by immunohistochemistry. Higher copy numbers of miR-J1 were detected in PML tissues than in non-PML tissues by real-time reverse transcription PCR. Next generation sequencing showed that miR-J1-5p, a mature miRNA of primary miRNA, was predominant in the lesions compared with miR-J1-3p, another mature miRNA. Deletion or mutation of miR-J1 in recombinant JCPyV promoted the production of JCPyV-encoded proteins in cells transfected with JCPyV DNA, suggesting that polyomavirus-encoded miRNA may have a repressive role in viral replication in PML tissues. In situ hybridization for viral miRNA may be a useful diagnostic tool for PML.


Assuntos
Vírus JC/genética , Leucoencefalopatia Multifocal Progressiva/genética , Adulto , Antígenos Virais de Tumores , Vírus BK/genética , DNA Viral/genética , Feminino , Expressão Gênica/genética , Regulação Viral da Expressão Gênica/genética , Genoma , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Oligodendroglia/metabolismo , Polyomavirus/genética , Infecções por Polyomavirus/genética , RNA Viral/genética , Transfecção , Carga Viral , Replicação Viral
5.
Nat Biomed Eng ; 4(6): 601-609, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32284553

RESUMO

In organ transplantation, infection and rejection are major causes of graft loss. They are linked by the net state of immunosuppression. To diagnose and treat these conditions earlier, and to improve long-term patient outcomes, refined strategies for the monitoring of patients after graft transplantation are needed. Here, we show that a fast and inexpensive assay based on CRISPR-Cas13 accurately detects BK polyomavirus DNA and cytomegalovirus DNA from patient-derived blood and urine samples, as well as CXCL9 messenger RNA (a marker of graft rejection) at elevated levels in urine samples from patients experiencing acute kidney transplant rejection. The assay, which we adapted for lateral-flow readout, enables-via simple visualization-the post-transplantation monitoring of common opportunistic viral infections and of graft rejection, and should facilitate point-of-care post-transplantation monitoring.


Assuntos
Sistemas CRISPR-Cas , Rejeição de Enxerto/virologia , Infecções Oportunistas/diagnóstico , Patologia Molecular/métodos , Biomarcadores/sangue , Biomarcadores/urina , Quimiocina CXCL9/sangue , Quimiocina CXCL9/urina , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Citomegalovirus/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , DNA Viral/sangue , DNA Viral/genética , DNA Viral/urina , Humanos , Rim , Nefropatias/virologia , Transplante de Rim/efeitos adversos , Masculino , Pessoa de Meia-Idade , Testes Imediatos , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/diagnóstico , Complicações Pós-Operatórias/diagnóstico , RNA Mensageiro , Infecções Tumorais por Vírus/diagnóstico
6.
Transplant Proc ; 52(3): 823-828, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32111385

RESUMO

BACKGROUND: Human polyoma virus-associated nephropathy frequently refers to allograft failure after kidney transplant. Thus, the early detection of viral activation is extremely important for these immunocompromised patients. METHODS: Previously, urine polyoma virus-infected cells (decoy cells) were indicated as the virus action, usually screened by the routine papanicolaou cytology in renal biopsy, but these methods are complex and the positive rate is low. In this article, the direct microscopy observation method, Wright-Giemsa staining, and Sternheimer-Malbin (SM) staining were all used to screen the decoy cells in urine samples of 213 kidney transplant patients who had used immunosuppressive drugs. RESULTS: Among them, decoy cells were detected in 40 cases (18.8%) by the direct observation method, 44 cases (20.7%) by Wright-Giemsa staining and 49 cases (23.0%) by SM staining. Furthermore, the most common polyoma viruses, BK and JC viruses, were also confirmed in 41 (83.7%) cases among these 49 decoy cell-positive samples. Importantly, compared with other decoy cell detection methods, SM staining is fast, easy to operate, and has a high positive rate. CONCLUSION: Therefore, SM staining is recommended as a fast and effective method for screening urine decoy cells in kidney transplant patients.


Assuntos
DNA Viral/urina , Nefropatias/diagnóstico , Infecções por Polyomavirus/diagnóstico , Polyomavirus/genética , Complicações Pós-Operatórias/diagnóstico , Coloração e Rotulagem/métodos , Adulto , Vírus BK/genética , Feminino , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/efeitos adversos , Vírus JC/genética , Nefropatias/patologia , Nefropatias/urina , Nefropatias/virologia , Transplante de Rim/efeitos adversos , Masculino , Teste de Papanicolaou , Infecções por Polyomavirus/urina , Infecções por Polyomavirus/virologia , Complicações Pós-Operatórias/urina , Complicações Pós-Operatórias/virologia , Transplante Homólogo , Urinálise/métodos , Ativação Viral
7.
J Virol ; 94(9)2020 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-32075934

RESUMO

Polyomaviruses (PyVs) are small DNA viruses carried by diverse vertebrates. The evolutionary relationships of viruses and hosts remain largely unclear due to very limited surveillance in sympatric communities. In order to investigate whether PyVs can transmit among different mammalian species and to identify host-switching events in the field, we conducted a systematic study of a large collection of bats (n = 1,083) from 29 sympatric communities across China which contained multiple species with frequent contact. PyVs were detected in 21 bat communities, with 192 PyVs identified in 186 bats from 15 species within 6 families representing at least 28 newly described PyVs. Surveillance results and phylogenetic analyses surprisingly revealed three interfamily PyV host-switching events in these sympatric bat communities: two distinct PyVs were identified in two bat species in restricted geographical locations, while another PyV clustered phylogenetically with PyVs carried by bats from a different host family. Virus-host relationships of all discovered PyVs were also evaluated, and no additional host-switching events were found. PyVs were identified in different horseshoe bat species in sympatric communities without observation of host-switching events, showed high genomic identities, and clustered with each other. This suggested that even for PyVs with high genomic identities in closely related host species, the potential for host switching is low. In summary, our findings revealed that PyV host switching in sympatric bat communities can occur but is limited and that host switching of bat-borne PyVs is relatively rare on the predominantly evolutionary background of codivergence with their hosts.IMPORTANCE Since the discovery of murine polyomavirus in the 1950s, polyomaviruses (PyVs) have been considered highly host restricted in mammals. Sympatric bat communities commonly contain several different bat species in an ecological niche facilitating viral transmission, and they therefore represent a model to identify host-switching events of PyVs. In this study, we screened PyVs in a large number of bats in sympatric communities from diverse habitats across China. We provide evidence that cross-species bat-borne PyV transmission exists, though is limited, and that host-switching events appear relatively rare during the evolutionary history of these viruses. PyVs with close genomic identities were also identified in different bat species without host-switching events. Based on these findings, we propose an evolutionary scheme for bat-borne PyVs in which limited host-switching events occur on the background of codivergence and lineage duplication, generating the viral genetic diversity in bats.


Assuntos
Quirópteros/genética , Quirópteros/virologia , Polyomavirus/genética , Animais , Evolução Biológica , China , Variação Genética/genética , Genoma Viral/genética , Especificidade de Hospedeiro/genética , Infecções por Polyomavirus/virologia , Análise de Sequência de DNA/métodos
8.
Virology ; 543: 27-33, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32056844

RESUMO

When purified from persistent infections, the genomes of most human polyomaviruses contain single enhancers. However, when isolated from productively infected cells from immunocompromised individuals, the genomes of several polyomaviruses contain duplicated enhancers that promote a number of polyoma-based diseases. The mechanism(s) that gives rise to the duplicated enhancers in the polyomaviruses is, however, not known. Herein we propose a model for the duplication of the enhancers that is based on recent advances in our understanding of; 1) the initiation of polyomavirus DNA replication, 2) the formation of long flaps via displacement synthesis and 3) the subsequent generation of duplicated enhancers via double stranded break repair. Finally, we discuss the possibility that the polyomavirus based replication dependent enhancer duplication model may be relevant to the enhancer-associated rearrangements detected in human genomes that are associated with various diseases, including cancers.


Assuntos
Reparo do DNA/genética , DNA Viral/biossíntese , Polyomavirus/genética , Replicação Viral/genética , Animais , DNA/genética , Quebras de DNA de Cadeia Dupla , Elementos de DNA Transponíveis , Elementos Facilitadores Genéticos , Humanos , Modelos Genéticos , Infecções por Polyomavirus/virologia
9.
mSphere ; 5(1)2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31941820

RESUMO

Diana V. Pastrana works in the field of DNA tumor virus biology. In this mSphere of Influence article, she reflects on how the two papers "Donor origin of BKV replication after kidney transplantation" (C. Schmitt, L. Raggub, S. Linnenweber-Held, O. Adams, et al., J Clin Virol 59:120-125, 2014, https://doi.org/10.1016/j.jcv.2013.11.009) and "Neutralizing antibody-mediated response and risk of BK virus-associated nephropathy" (M. Solis, A. Velay, R. Porcher, P. Domingo-Calap, et al., J Am Soc Nephrol 29:326-334, 2018, https://doi.org/10.1681/ASN.2017050532) reminded her of the importance of allowing data, and not adherence to dogma, to drive her research.


Assuntos
Transplante de Rim/efeitos adversos , Doadores de Tecidos , DNA Viral/genética , Feminino , Humanos , Imunossupressores , Polyomavirus/genética , Infecções por Polyomavirus/prevenção & controle
10.
Int J Pharm ; 576: 119008, 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-31901358

RESUMO

Viral nanoparticles represent potential natural versatile platforms for targeted gene and drug delivery. Improving the efficiency of gene transfer mediated by viral vectors could not only enhance their therapeutic potential, but also contribute to understanding the limitations in interactions of nanoparticles with cells and the development of new therapeutic approaches. In this study, four cell-penetrating peptides (CPPs), cationic octaarginine (R8), histidine-rich peptides (LAH4 and KH27K) and fusogenic peptide (FUSO), are investigated for their effect on infection by mouse polyomavirus (MPyV) or on transduction of reporter genes delivered by MPyV or related viral vectors. Peptides noncovalently associated with viral particles enhance gene transfer (with the exception of FUSO). Removal of cellular heparan sulfates by the heparinase does not significantly change the enhancing potential of CPPs. Instead, CPPs influences the physical state of viral particles: R8 slightly destabilizes the intact virus, KH27K induces its aggregation and LAH4 promotes disassembly and aggregation of the particles that massively and rapidly associate with cells. The findings indicate that peptides acting as transduction-enhancing agents of polyomavirus-based nanoparticles modulate their physical state, which can be an important prerequisite for sensitization of cells and determination of the further fate of viral particles inside cells.


Assuntos
Peptídeos Penetradores de Células/metabolismo , Vetores Genéticos , Polyomavirus/metabolismo , Transdução Genética , Vírion/metabolismo , Animais , Capsídeo/metabolismo , Capsídeo/ultraestrutura , Peptídeos Penetradores de Células/química , Células HEK293 , Humanos , Camundongos , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Polyomavirus/genética , Polyomavirus/ultraestrutura , Vírion/genética , Vírion/ultraestrutura
11.
Jpn J Infect Dis ; 73(2): 132-139, 2020 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-31787742

RESUMO

Trichodysplasia spinulosa-associated polyomavirus (TSPyV or human polyomavirus 8) was identified from patients with trichodysplasia spinulosa, a rare skin disease affecting the faces of immunocompromised patients. Like other polyomaviruses, the TSPyV genome encodes a large T antigen (LT). However, the expression and functions of TSPyV LT in infected cells remain largely unknown. In the present study, we cloned a full-length TSPyV LT cDNA from cells transfected with the full-length of TSPyV LT DNA. Transfection study using green fluorescence protein-tagged LT expression plasmids showed that TSPyV LT was expressed in the nucleus of transfected cells. Analysis of deletion mutants identified a nuclear localization signal in TSPyV LT. Recombinant TSPyV LT exhibited an ATPase activity. TSPyV LT has a chitinase-like domain; however, no chitinase activity was detected. Immunoprecipitation assays revealed that TSPyV LT bound to retinoblastoma 1, but not to p53 in transfected cells. Expression of TSPyV LT in NIH3T3 cells induced colony formation in soft agar, suggesting its transformation activity. These data indicate that TSPyV LT may be associated with the pathogenesis of trichodysplasia spinulosa, which is a hyperplasia of keratinocytes in inner hair follicles.


Assuntos
Antígenos Virais de Tumores/genética , Antígenos Virais de Tumores/metabolismo , Polyomavirus/genética , Dermatopatias/virologia , Adenosina Trifosfatases/metabolismo , Animais , Hiperplasia , Hospedeiro Imunocomprometido , Queratinócitos/patologia , Queratinócitos/virologia , Camundongos , Células NIH 3T3 , Sinais de Localização Nuclear/genética , Polyomavirus/patogenicidade , Infecções por Polyomavirus/virologia , Dermatopatias/patologia
13.
Virol J ; 16(1): 137, 2019 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-31727090

RESUMO

BACKGROUND: Polyomaviruses (PyVs) have a wide range of hosts, from humans to fish, and their effects on hosts vary. The differences in the infection characteristics of PyV with respect to the host are assumed to be influenced by the biochemical function of the LT-Ag protein, which is related to the cytopathic effect and tumorigenesis mechanism via interaction with the host protein. METHODS: We carried out a comparative analysis of codon usage patterns of large T-antigens (LT-Ags) of PyVs isolated from various host species and their functional domains and sequence motifs. Parity rule 2 (PR2) and neutrality analysis were applied to evaluate the effects of mutation and selection pressure on codon usage bias. To investigate evolutionary relationships among PyVs, we carried out a phylogenetic analysis, and a correspondence analysis of relative synonymous codon usage (RSCU) values was performed. RESULTS: Nucleotide composition analysis using LT-Ag gene sequences showed that the GC and GC3 values of avian PyVs were higher than those of mammalian PyVs. The effective number of codon (ENC) analysis showed host-specific ENC distribution characteristics in both the LT-Ag gene and the coding sequences of its domain regions. In the avian and fish PyVs, the codon diversity was significant, whereas the mammalian PyVs tended to exhibit conservative and host-specific evolution of codon usage bias. The results of our PR2 and neutrality analysis revealed mutation bias or highly variable GC contents by showing a narrow GC12 distribution and wide GC3 distribution in all sequences. Furthermore, the calculated RSCU values revealed differences in the codon usage preference of the LT-AG gene according to the host group. A similar tendency was observed in the two functional domains used in the analysis. CONCLUSIONS: Our study showed that specific domains or sequence motifs of various PyV LT-Ags have evolved so that each virus protein interacts with host cell targets. They have also adapted to thrive in specific host species and cell types. Functional domains of LT-Ag, which are known to interact with host proteins involved in cell proliferation and gene expression regulation, may provide important information, as they are significantly related to the host specificity of PyVs.


Assuntos
Antígenos Virais de Tumores/genética , Uso do Códon , Infecções por Polyomavirus/veterinária , Infecções por Polyomavirus/virologia , Polyomavirus/genética , Motivos de Aminoácidos , Animais , Composição de Bases , Aves , Biologia Computacional , Peixes , Humanos , Mamíferos , Filogenia , Polyomavirus/isolamento & purificação
14.
Vet Microbiol ; 237: 108397, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31585638

RESUMO

Aves polyomavirus 1 (APV) causes inflammatory disease in psittacine birds, especially in young budgerigar. In this study, an APV virus (SD18 strain) was isolated from a diseased psittacine birds breeding facility. The full genome (4981 bp) of SD18 was determined and analyzed. Phylogenetic analysis of full genome sequences indicated all the APV strains form two groups. The SD18 strain showed close relationship with APV isolated from Poland, however, the other Chinese strains are located in group II, which suggested different genotypes APVs are co-circulating in China. Compared with the consensus sequence of APV full genome, the SD18 strain contains 13 nucleotide mutations, and 2 unique amino acid substitutions (R179M and Q382K) located in VP2/3 and Large T proteins. To explore the pathogenicity of the virus, the SD18 strain was used to challenge 2-week-old budgerigars. All infected birds died no later than 5 days post infection, and virus was detected in multiple organs including brain, heart, ingluvies, liver, and intestine, which indicated that SD18 is fatal and causes systemic infection in young budgerigar. In vitro studies showed that SD18 replicated efficiently in CEF cells and reached the highest viral titers at 9 days post infection. Notably, replication of SD18 stimulated IFN-ß response in CEF cells and overexpression of the VP4 or VP4Delta proteins significantly inhibited IFN-ß promoter activation, which could be the strategy of APV to escape from the host innate immunity.


Assuntos
Doenças das Aves/virologia , Melopsittacus/virologia , Infecções por Polyomavirus/veterinária , Polyomavirus/isolamento & purificação , Animais , Doenças das Aves/epidemiologia , Surtos de Doenças/veterinária , Genoma Viral , Filogenia , Polyomavirus/genética , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/virologia
15.
Viruses ; 11(10)2019 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-31658738

RESUMO

As the phylogenetic organization of mammalian polyomaviruses is complex and currently incompletely resolved, we aimed at a deeper insight into their evolution by identifying polyomaviruses in host orders and families that have either rarely or not been studied. Sixteen unknown and two known polyomaviruses were identified in animals that belong to 5 orders, 16 genera, and 16 species. From 11 novel polyomaviruses, full genomes could be determined. Splice sites were predicted for large and small T antigen (LTAg, STAg) coding sequences (CDS) and examined experimentally in transfected cell culture. In addition, splice sites of seven published polyomaviruses were analyzed. Based on these data, LTAg and STAg annotations were corrected for 10/86 and 74/86 published polyomaviruses, respectively. For 25 polyomaviruses, a spliced middle T CDS was observed or predicted. Splice sites that likely indicate expression of additional, alternative T antigens, were experimentally detected for six polyomaviruses. In contrast to all other mammalian polyomaviruses, three closely related cetartiodactyl polyomaviruses display two introns within their LTAg CDS. In addition, the VP2 of Glis glis (edible dormouse) polyomavirus 1 was observed to be encoded by a spliced transcript, a unique experimental finding within the Polyomaviridae family. Co-phylogenetic analyses based on LTAg CDS revealed a measurable signal of codivergence when considering all mammalian polyomaviruses, most likely driven by relatively recent codivergence events. Lineage duplication was the only other process whose influence on polyomavirus evolution was unambiguous. Finally, our analyses suggest that an update of the taxonomy of the family is required, including the creation of novel genera of mammalian and non-mammalian polyomaviruses.


Assuntos
Antígenos Virais de Tumores/genética , Mamíferos/virologia , Polyomavirus , Animais , Evolução Biológica , Classificação , Genes Virais , Genoma Viral , Humanos , Filogenia , Polyomavirus/classificação , Polyomavirus/genética , Polyomavirus/isolamento & purificação
16.
Transfusion ; 59(12): 3689-3697, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31633816

RESUMO

BACKGROUND: Human polyomaviruses (HPyVs), like herpesviruses, cause persistent infection in a large part of the population. In immunocompromised and elderly patients, PyVs cause severe diseases such as nephropathy (BK polyomavirus [BKPyV]), progressive multifocal leukoencephalopathy (JC polyomavirus [JCPyV]), and skin cancer (Merkel cell polyomavirus [MCPyV]). Like cytomegalovirus, donor-derived PyV can cause disease in kidney transplant recipients. Possibly blood components transmit PyVs as well. To study this possibility, as a first step we determined the presence of PyV DNA in Dutch blood donations. STUDY DESIGN AND METHODS: Blood donor serum samples (n = 1016) were analyzed for the presence of DNA of 14 HPyVs using HPyV species-specific quantitative polymerase chain reaction (PCR) procedures. PCR-positive samples were subjected to confirmation by sequencing. Individual PCR findings were compared with the previously reported PyV serostatus. RESULTS: MC polyomavirus DNA was detected in 39 donors (3.8%), JCPyV and TS polyomavirus (TSPyV) DNA in five donors (both 0.5%), and HPyV9 DNA in four donors (0.4%). BKPyV, WU polyomavirus (WUPyV), HPyV6, MW polyomavirus (MWPyV), and LI polyomavirus (LIPyV) DNA was detected in one or two donors. Amplicon sequencing confirmed the expected product for BKPyV, JCPyV, WUPyV, MCPyV, HPyV6, TSPyV, MWPyV, HPyV9, and LIPyV. For JCPyV a significant association was observed between detection of viral DNA and the level of specific IgG antibodies. CONCLUSION: In 5.4% of Dutch blood donors PyV DNA was detected, including DNA from pathogenic PyVs such as JCPyV. As a next step, the infectivity of PyV in donor blood and transmission via blood components to immunocompromised recipients should be investigated.


Assuntos
Doadores de Sangue/estatística & dados numéricos , DNA Viral/análise , Polyomavirus/genética , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polyomavirus/isolamento & purificação , Polyomavirus/patogenicidade , Prevalência , Adulto Jovem
17.
Arch Virol ; 164(11): 2837-2841, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31494776

RESUMO

Since January 2019, abnormal molting has been observed frequently in approximately 40-day-old Pekin ducks in China. To investigate the possible involvement of a virus, we tested the prevalence of duck circovirus (DuCV), goose hemorrhagic polyomavirus (GHPyV), and goose parvovirus (GPV) in 11 molt cases in two provinces. GPV was detected in all cases, particularly in all samples collected from the feather area. The complete genome sequences of three GPV strains were determined and found to have 52 nucleotide changes relative to GPVs associated with short beak and dwarfism syndrome of Pekin ducks. These data will enhance our understanding of GPV diversity and outcomes of GPV infection in Pekin ducks.


Assuntos
Patos/virologia , Gansos/virologia , Muda/fisiologia , Parvovirinae/isolamento & purificação , Doenças das Aves Domésticas/virologia , Animais , China/epidemiologia , Circovirus/genética , Circovirus/isolamento & purificação , Genoma Viral/genética , Parvovirinae/genética , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Doenças das Aves Domésticas/epidemiologia
18.
Med Microbiol Immunol ; 208(6): 845-854, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31375897

RESUMO

Fragmented data are available on the human polyomaviruses (HPyVs) prevalence in the gastrointestinal tract. Rearrangements in the non-coding control region (NCCR) of JCPyV and BKPyV have been extensively studied and correlated to clinical outcome; instead, little information is available for KIPyV, WUPyV and MCPyV NCCRs. To get insights into the role of HPyVs in the gastrointestinal tract, we investigated JCPyV, BKPyV, KIPyV, WUPyV and MCPyV distribution among hematological patients in concomitance with gastrointestinal symptoms. In addition, NCCRs and VP1 sequences were examined to characterize the strains circulating among the enrolled patients. DNA was extracted from 62 stool samples and qPCR was carried out to detect and quantify JCPyV, BKPyV, KIPyV, WUPyV and MCPyV genomes. Positive samples were subsequently amplified and sequenced for NCCR and VP1 regions. A phylogenetic tree was constructed aligning the obtained VP1 sequences to a set of reference sequences. qPCR revealed low viral loads for all HPyVs searched. Mono and co-infections were detected. A significant correlation was found between gastrointestinal complications and KIPyV infection. Archetype-like NCCRs were found for JCPyV and BKPyV, and a high degree of NCCRs stability was observed for KIPyV, WUPyV and MCPyV. Analysis of the VP1 sequences revealed a 99% identity with the VP1 reference sequences. The study adds important information on HPyVs prevalence and persistence in the gastrointestinal tract. Gastrointestinal signs were correlated with the presence of KIPyV, although definitive conclusions cannot be drawn. HPyVs NCCRs showed a high degree of sequence stability, suggesting that sequence rearrangements are rare in this anatomical site.


Assuntos
Fezes/virologia , Variação Genética , Neoplasias Hematológicas/complicações , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/virologia , Polyomavirus/isolamento & purificação , Eliminação de Partículas Virais , Adulto , Criança , Feminino , Humanos , Masculino , Filogenia , Polyomavirus/classificação , Polyomavirus/genética , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Carga Viral
19.
J Vet Diagn Invest ; 31(5): 719-725, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31423916

RESUMO

Aves polyomavirus 1, psittacine beak and feather disease virus, and psittacid herpesvirus 1 are important pathogens of psittacine birds with the potential to cause substantial morbidity and mortality. Using publically available nucleotide sequences, we developed and validated a triplex real-time PCR (rtPCR) assay to rapidly detect these 3 viruses. The assay had high analytical sensitivity, detecting <6 copies of viral DNA per reaction, and 100% analytical specificity, showing no cross-reactivity with 59 other animal pathogens. Archived formalin-fixed, paraffin-embedded tissues from psittacine birds diagnosed at postmortem as infected with each of the viruses as well as virus-negative birds were used to validate the utility of the assay. Birds were selected for the positive cohort if they showed histologic evidence of infection (i.e., characteristic inclusion bodies in tissues); birds in the negative cohort had final diagnoses unrelated to the pathogens of interest. The triplex rtPCR assay confirmed 98% of histopathology-positive cases, and also identified subclinical infections that were not observed by histologic examination, including coinfections. Birds that tested positive only by rtPCR had significantly higher cycle threshold values compared to those with histologic evidence of infection. Positive, negative, and overall percentage agreements as well as the kappa statistic between the results of the assay and histopathology were high, demonstrating the usefulness of the assay as a tool to confirm disease diagnoses, and to improve detection of subclinical infections.


Assuntos
Doenças das Aves/diagnóstico , Infecções por Vírus de DNA/veterinária , Vírus de DNA/isolamento & purificação , Herpesviridae/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/veterinária , Psittaciformes/virologia , Alphaherpesvirinae/genética , Alphaherpesvirinae/isolamento & purificação , Animais , Doenças das Aves/virologia , Infecções por Circoviridae/diagnóstico , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Circovirus/genética , Circovirus/isolamento & purificação , Infecções por Vírus de DNA/diagnóstico , Infecções por Vírus de DNA/virologia , Vírus de DNA/genética , DNA Viral , Herpesviridae/genética , Papagaios/virologia , Polyomaviridae/genética , Polyomaviridae/isolamento & purificação , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária
20.
Nat Commun ; 10(1): 3425, 2019 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-31366885

RESUMO

Single-stranded (ss) DNA viruses are a major component of the earth virome. In particular, the circular, Rep-encoding ssDNA (CRESS-DNA) viruses show high diversity and abundance in various habitats. By combining sequence similarity network and phylogenetic analyses of the replication proteins (Rep) belonging to the HUH endonuclease superfamily, we show that the replication machinery of the CRESS-DNA viruses evolved, on three independent occasions, from the Reps of bacterial rolling circle-replicating plasmids. The CRESS-DNA viruses emerged via recombination between such plasmids and cDNA copies of capsid genes of eukaryotic positive-sense RNA viruses. Similarly, the rep genes of prokaryotic DNA viruses appear to have evolved from HUH endonuclease genes of various bacterial and archaeal plasmids. Our findings also suggest that eukaryotic polyomaviruses and papillomaviruses with dsDNA genomes have evolved via parvoviruses from CRESS-DNA viruses. Collectively, our results shed light on the complex evolutionary history of a major class of viruses revealing its polyphyletic origins.


Assuntos
Archaea/genética , Bactérias/genética , DNA Viral/genética , Evolução Molecular , Plasmídeos/genética , Sequência de Bases , DNA Helicases/genética , DNA de Cadeia Simples/genética , Genoma Viral/genética , Papillomaviridae/genética , Parvovirus/genética , Polyomavirus/genética , Alinhamento de Sequência
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...