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1.
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
2.
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
3.
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
4.
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
5.
Vet Res Commun ; 43(4): 197-202, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31297735

RESUMO

Avian polyomavirus (APV) infection, also called as budgerigar fledgling disease (BFD) causes various health problems in many psittacine species which may cause untimely death. The aims of this study were to investigate, for the first time, the detection, molecular characterization and phylogenetic analysis of avian polyomavirus (APV) in Pakistani psittacine birds. In an aviary a disease similar to APV was found and 90% of the nestlings died within a few weeks. Seven to ten-day-old parrot nestlings (n = 3) from the aviary were presented with feather abnormalities, plumage defect and were clinically depressed. Birds died at 11th, 14th and 16th day of age. Samples of hearts, livers, spleen, feathers and kidneys were collected from the dead birds. Samples were analyzed for the presence of APV DNA by using PCR. APV VP1 gene was partially sequenced, and phylogenetic analysis was performed. The APV strain was similar to those previously reported in other areas of the world. The results of this investigation indicate presence of a high frequency of APV infections in psittacine birds in Pakistan.


Assuntos
Doenças das Aves/virologia , Papagaios/virologia , Infecções por Polyomavirus/veterinária , Polyomavirus/classificação , Polyomavirus/genética , Animais , Doenças das Aves/diagnóstico , Doenças das Aves/patologia , Proteínas do Capsídeo/genética , Paquistão , Filogenia , Reação em Cadeia da Polimerase , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/patologia , Infecções por Polyomavirus/virologia
6.
Artigo em Inglês | MEDLINE | ID: mdl-31275867

RESUMO

An effective vaccine against the Plasmodium parasite is likely to require the induction of robust antibody and T cell responses. Chimeric virus-like particles are an effective vaccine platform for induction of antibody responses, but their capacity to induce robust cellular responses and cell-mediated protection against pathogen challenge has not been established. To evaluate this, we produced chimeric constructs using the murine polyomavirus structural protein with surface-exposed CD8+ or CD4+ T cell or B cell repeat epitopes derived from the Plasmodium yoelii circumsporozoite protein, and assessed immunogenicity and protective capacity in a murine model. Robust CD8+ T cell responses were induced by immunization with the chimeric CD8+ T cell epitope virus-like particles, however CD4+ T cell responses were very low. The B cell chimeric construct induced robust antibody responses but there was no apparent synergy when T cell and B cell constructs were administered as a pool. A heterologous prime/boost regimen using plasmid DNA priming followed by a VLP boost was more effective than homologous VLP immunization for cellular immunity and protection. These data show that chimeric murine polyomavirus virus-like particles are a good platform for induction of CD8+ T cell responses as well as antibody responses.


Assuntos
Formação de Anticorpos/imunologia , Antígenos de Protozoários/imunologia , Linfócitos T CD8-Positivos/imunologia , Polyomavirus/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia , Animais , Anticorpos Antiprotozoários , Linfócitos B , Linfócitos T CD4-Positivos , Modelos Animais de Doenças , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Imunidade Celular , Imunização , Imunização Secundária , Vacinas Antimaláricas , Camundongos , Camundongos Endogâmicos BALB C , Plasmodium yoelii , Polyomavirus/genética , Proteínas de Protozoários/imunologia , Vacinas de Partículas Semelhantes a Vírus/genética
7.
Virol J ; 16(1): 35, 2019 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-30885224

RESUMO

BACKGROUND: Papillomaviruses (PVs) and polyomaviruses (PyVs) infect diverse vertebrates including human and cause a broad spectrum of outcomes from asymptomatic infection to severe disease. There has been no PV and only one PyV detected in tree shrews, though the genomic properties of tree shrews are highly similar to those of the primates. METHODS: Swab and organ samples of tree shrews collected in the Yunnan Province of China, were tested by viral metagenomic analysis and random PCR to detect the presence of PVs and PyVs. By PCR amplification using specific primers, cloning, sequencing and assembling, genomes of two PVs and one PyV were identified in the samples. RESULTS: Two novel PVs and a novel PyV, named tree shrew papillomavirus 1 and 2 (TbelPV1 and TbelPV2) and polyomavirus 1 (TbelPyV1) were characterized in the Chinese tree shrew (Tupaia belangeri chinensis). The genomes of TbelPV1, TbelPV2, and TbelPyV1 are 7410 bp, 7526 bp, and 4982 bp in size, respectively. The TbelPV1 genome contains 7 putative open-reading frames (ORFs) coding for viral proteins E1, E2, E4, E6, E7, L1, and L2; the TbelPV2 genome contains 6 ORFs coding for viral proteins E1, E2, E6, E7, L1, and L2; and the TbelPyV1 genome codes for the typical small and large T antigens of PyV, as well as the VP1, VP2, and VP3 capsid proteins. Genomic comparison and phylogenetic analysis indicated that TbelPV1 and TbelPV2 represented 2 novel PV genera of Papillomaviridae, and TbelPyV1 represented a new species of genus Alphapolyomavirus. Our epidemiologic study indicated that TbelPV1 and TbelPV2 were both detected in oral swabs, while TbelPyV1 was detected in oral swabs and spleens. CONCLUSION: Two novel PVs (TbelPV1 and TbelPV2) and a novel PyV (TbelPyV) were discovered in tree shrews and their genomes were characterized. TbelPV1, TbelPV2, and TbelPyV1 have the highest similarity to Human papillomavirus type 63, Ursus maritimus papillomavirus 1, and Human polyomavirus 9, respectively. TbelPV1 and TbelPV2 only showed oral tropism, while TbelPyV1 showed oral and spleen tropism.


Assuntos
Genoma Viral , Papillomaviridae/genética , Polyomavirus/genética , Tupaia/virologia , Animais , China , Genômica , Metagenômica , Boca/virologia , Fases de Leitura Aberta , Filogenia , Reação em Cadeia da Polimerase , Baço/virologia , Proteínas Virais/genética , Tropismo Viral
8.
J Fish Dis ; 42(3): 345-355, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30632177

RESUMO

In Taiwan, a petechial haemorrhage disease associated with mortality has affected marbled eels (Anguilla marmorata). The eels were revealed to be infected with adomavirus (MEAdoV, previously recognized as a polyoma-like virus). In this study, cell line DMEPF-5 was established from the pectoral fin of a diseased eel. DMEPF-5 was passaged >70 times and thoroughly proliferated in L-15 medium containing 2%-15% foetal bovine serum at 20-30°C. Transcripts of neural cell adhesion molecule 1 and nestin genes, and nucleic acids of MEAdoV and a novel reovirus (MERV) in the cells were demonstrated by reverse transcription-polymerase chain reaction analysis. Phylogenetic analysis revealed that the AdoV LO8 proteins mostly relate to adenovirus adenain, whereas MERV is close to American grass carp reovirus in Aquareovirus G, based on a partial VP2 nucleotide sequence. DMEPF-5 cells are susceptible to additional viral infection. Taken together, the marbled eels with the haemorrhagic disease have coinfection with MEAdoV and MERV, and the pathogenic role of MEAdoV and MERV warrants research. DMEPF-5 has gene expression associated with mesenchymal stem and progenitor cells and is the first cell line persistently infected with adomavirus and aquareovirus. DMEPF-5 can facilitate studies of such viruses and haemorrhagic disease.


Assuntos
Anguilla , Linhagem Celular/virologia , Doenças dos Peixes/virologia , Infecções por Polyomavirus/veterinária , Infecções por Reoviridae/veterinária , Sequência de Aminoácidos , Nadadeiras de Animais/citologia , Nadadeiras de Animais/virologia , Animais , Sequência de Bases , Suscetibilidade a Doenças/veterinária , Suscetibilidade a Doenças/virologia , Polyomavirus/genética , Infecções por Polyomavirus/virologia , Púrpura/veterinária , Púrpura/virologia , Reoviridae/genética , Infecções por Reoviridae/virologia , Pele/patologia , Pele/virologia
9.
Braz J Microbiol ; 50(1): 133-137, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30637634

RESUMO

In 2007, the new polyomaviruses WUPyV and KIPyV were identified in patients with acute respiratory infections. The aim of this study was to investigate these viruses in hospitalized patients with severe acute respiratory infection (SARI). A retrospective study was conducted with 251 patients, from April 2009 to November 2010, using nasopharyngeal aspirates, naso- and oropharyngeal swab samples from hospitalized patients (children < 12 years and adults) who had SARI within 7 days of the onset of symptoms, including fever (> 38.8 °C), dyspnea, and cough. Clinical and epidemiological information was obtained through standardized questionnaire. Enrolled patients were initially suspected to have influenza A(H1N1)pdm09 infections. WUPyV and KIPyV were detected by real-time PCR. Samples were also tested for influenza A and B viruses, human respiratory syncytial virus, rhinovirus, metapneumovirus, coronavirus, adenovirus, and parainfluenza viruses. WUPyV and KIPyV were detected in 6.77% (4.78% and 1.99%, respectively) of hospitalized patients with SARI. All samples from children showed coinfections (rhinovirus was the most commonly detected). Six adults had polyomavirus infection and four (1.6%) had monoinfection. Of them, 3 reported comorbidities including immunosuppression and 1 patient had worse outcome, requiring ICU admission. These preliminary data may suggest a possible role of polyomaviruses in SARI among immunocompromised adult patients.


Assuntos
Infecções por Polyomavirus/virologia , Polyomavirus/isolamento & purificação , Infecções Respiratórias/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Polyomavirus/classificação , Polyomavirus/genética , Adulto Jovem
10.
Eur J Clin Microbiol Infect Dis ; 38(1): 135-139, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30338464

RESUMO

Despite the growing importance of infections caused by the human polyomaviruses (HPyVs), information about their transmission, pathogenesis, and epidemiology is scarce. The objective of this work was to evaluate the excretion and distribution of HPyV (HPyV1-HPyV4 [former BKPyV, JCPyV, KIPyV, and WUPyV, respectively]) among asymptomatic individuals from different geographic regions in Brazil, in order to verify the existence of distinct epidemiologic patterns among the Brazilian population. Saliva samples from 889 healthy volunteers living in nine locations in Brazil were analyzed by real-time polymerase chain reaction (PCR) to detect HPyV1-4. Among 889 participants, 346 (39%) had evidence of infection with one or more HPyV species: 127 (14.3%) had HPyV1 only; 70 (7.9%) had HPyV3 only; 60 (6.7%) had HPyV4 only, and 25 (2.8%) had HPyV2 only. Coinfections were detected in 64 participants (7.3%). Although HPyV excretion was detected in samples from all locations, the frequency and distribution of viral species varied significantly. The epidemiologic findings presented demonstrate that the four HPyV species studied are circulating in five geographic regions of Brazil. Salivary excretion of these viruses appears common among healthy Brazilians. The distribution of viral species varies considerably between regions as well as within regions.


Assuntos
Infecções por Polyomavirus/epidemiologia , Polyomavirus/genética , Adolescente , Adulto , Idoso , Infecções Assintomáticas/epidemiologia , Brasil/epidemiologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Infecções por Polyomavirus/virologia , Saliva/virologia , Adulto Jovem
11.
J Invest Dermatol ; 139(2): 285-292, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30470393

RESUMO

Human polyomaviruses are double-stand DNA viruses with a conserved genomic structure, yet they present with diverse tissue tropisms and disease presentations. Merkel cell polyomavirus, trichodysplasia spinulosa polyomavirus, human polyomavirus 6 and 7, and Malawi polyomavirus are shed from the skin, and Merkel cell polyomavirus, trichodysplasia spinulosa polyomavirus, human polyomavirus 6 and 7 have been linked to specific skin diseases. We present an update on the genomic and clinical features of these cutaneous polyomaviruses.


Assuntos
Infecções por Polyomavirus/diagnóstico , Polyomavirus/genética , Dermatopatias Virais/diagnóstico , Antígenos Virais/genética , Genoma Viral/genética , Humanos , Polyomavirus/imunologia , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/imunologia , Infecções por Polyomavirus/virologia , Pele/imunologia , Pele/patologia , Pele/virologia , Dermatopatias Virais/imunologia , Dermatopatias Virais/virologia
12.
Microbiol Immunol ; 62(12): 763-773, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30537287

RESUMO

Trichodysplasia spinulosa-associated polyomavirus (TSPyV), a newly identified polyomavirus, has been implicated as a causative agent of trychodysplasia spinulosa (TS), a rare proliferative skin disease in severely immunocompromised hosts. Diagnosis using mAbs is a promising tool with high specificity towards the specific antigen. However, thus far, no suitable mAbs for diagnosing TS disease have been identified. In this study, mAbs specific for VP1 of TSPyV were developed and characterized. Wheat germ cell-free synthesized VP1 protein of TSPyV was used to immunize BALB/c mice to generate hybridomas. Screening of the resultant hybridoma clones resulted in selection of five strongly positive clones that produce mAbs that react with the TSPyV-VP1 antigen. Epitope mapping and bioinformatic analysis showed that these mAbs recognized epitopes located within highly conserved C-terminal region of all clinical isolates of TSPyV-VP1. Further, all these mAbs were highly effective for immunofluorescence and immunoprecipitation analysis. Three of the five mAbs exhibited no cross-reactivity with VP1 of other related polyomaviruses. In addition, one of our mAbs (#14) provided immunohistochemical staining of skin tissue of TS disease. It can be concluded that three of the mAbs in this panel of anti-VP1 antibodies may provide a useful set of tools for studying TSPyV infection and making the specific diagnosis.


Assuntos
Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/isolamento & purificação , Proteínas do Capsídeo/imunologia , Infecções por Polyomavirus/imunologia , Polyomavirus/imunologia , Infecções Tumorais por Vírus/imunologia , Animais , Proteínas do Capsídeo/genética , DNA Viral , Modelos Animais de Doenças , Mapeamento de Epitopos , Epitopos/imunologia , Feminino , Regulação Viral da Expressão Gênica , Genes Virais/genética , Humanos , Hibridomas , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Modelos Moleculares , Polyomavirus/genética , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/virologia , Alinhamento de Sequência , Pele/patologia , Infecções Tumorais por Vírus/diagnóstico
13.
Future Microbiol ; 13: 1719-1730, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30484707

RESUMO

AIM: To study prevalence of Karolinska Institutet (KI) and Washington University (WU) polyomavirus (PyV) in 100 tonsils, 100 adenoids, 146 throat swab and 15 middle ear fluid samples collected from 146 patients (120 children and 26 adults), to analyze the sequence of  noncoding control region (NCCR) and complete WUPyV genomes. MATERIALS & METHODS: Viruses were detected by quantitative real-time PCR. The NCCRs and WUPyV genomes were sequenced and analyzed. RESULTS: The frequency of WUPyV and KIPyV DNA was 27 and 11% in adenoids, 4 and 3% in tonsils, 4.1 and 1.4% in throat swab samples, respectively. The WUPyV DNA was detected in one middle ear fluid sample as well. The WUPyV NCCRs showed mutations which may alter the putative transcription factor binding sites. Phylogenetic analysis revealed three clades of WUPyV. CONCLUSION: Tonsils and adenoids might be site of virus replication and/or persistence, and WUPyV may invade into the middle ear.


Assuntos
Tonsila Faríngea/virologia , Orelha Média/virologia , Genoma Viral/genética , Tonsila Palatina/virologia , Faringe/virologia , Polyomavirus/genética , Adulto , Criança , Pré-Escolar , DNA Viral/genética , Feminino , Humanos , Masculino , Filogenia , Polyomavirus/isolamento & purificação , Infecções por Polyomavirus/virologia , Infecções Respiratórias/virologia , Sequenciamento Completo do Genoma
14.
Breast Cancer Res ; 20(1): 131, 2018 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-30367629

RESUMO

BACKGROUND: Amphiregulin (AREG), a ligand of the epidermal growth factor receptor, is not only essential for proper mammary ductal development, but also associated with breast cancer proliferation and growth. In the absence of AREG, mammary ductal growth is stunted and fails to expand. Furthermore, suppression of AREG expression in estrogen receptor-positive breast tumor cells inhibits in-vitro and in-vivo growth. METHODS: We crossed AREG-null (AREG-/-) mice with the murine luminal B breast cancer model, MMTV-PyMT (PyMT), to generate spontaneous breast tumors that lack AREG (AREG-/- PyMT). We evaluated tumor growth, cytokeratin-8 (K8)-positive luminal cells, cytokeratin-14 (K14)-positive myoepithelial cells, and expression of AREG, Ki67, and PyMT. Primary myoepithelial cells from nontumor-bearing AREG+/+ mice underwent fluorescence-activated cell sorting and were adapted to culture for in-vitro coculture studies with AT-3 cells, a cell line derived from C57Bl/6 PyMT mammary tumors. RESULTS: Intriguingly, PyMT-induced lesions progress more rapidly in AREG-/- mice than in AREG+/+ mice. Quantification of K8+ luminal and K14+ myoepithelial cells in non-PyMT AREG-/- mammary glands showed fewer K14+ cells and a thinner myoepithelial layer. Study of AT-3 cells indicated that coculture with myoepithelial cells or exposure to AREG, epidermal growth factor, or basic fibroblast growth factor can suppress PyMT expression. Late-stage AREG-/- PyMT tumors are significantly less solid in structure, with more areas of papillary and cystic growth. Papillary areas appear to be both less proliferative and less necrotic. In The Cancer Genome Atlas database, luminal-B invasive papillary carcinomas have lower AREG expression than luminal B invasive ductal carcinomas. CONCLUSIONS: Our study has revealed a previously unknown role of AREG in myoepithelial cell development and PyMT expression. AREG expression is essential for proper myoepithelial coverage of mammary ducts. Both AREG and myoepithelial cells can suppress PyMT expression. We find that lower AREG expression is associated with invasive papillary breast cancer in both the MMTV-PyMT model and human breast cancer.


Assuntos
Anfirregulina/metabolismo , Células Epiteliais/patologia , Glândulas Mamárias Animais/patologia , Neoplasias Mamárias Experimentais/patologia , Anfirregulina/genética , Animais , Antígenos Transformantes de Poliomavirus/genética , Antígenos Transformantes de Poliomavirus/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Células Epiteliais/virologia , Feminino , Humanos , Glândulas Mamárias Animais/citologia , Neoplasias Mamárias Experimentais/genética , Neoplasias Mamárias Experimentais/virologia , Vírus do Tumor Mamário do Camundongo/genética , Vírus do Tumor Mamário do Camundongo/patogenicidade , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Invasividade Neoplásica/patologia , Polyomavirus/genética , Polyomavirus/imunologia
15.
Nat Rev Clin Oncol ; 15(12): 763-776, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30287935

RESUMO

Merkel cell carcinoma (MCC) is a rare and aggressive skin cancer associated with advanced age and immunosuppression. Over the past decade, an association has been discovered between MCC and either integration of the Merkel cell polyomavirus, which likely drives tumorigenesis, or somatic mutations owing to ultraviolet-induced DNA damage. Both virus-positive and virus-negative MCCs are immunogenic, and inhibition of the programmed cell death protein 1 (PD-1)-programmed cell death 1 ligand 1 (PD-L1) immune checkpoint has proved to be highly effective in treating patients with metastatic MCC; however, not all patients have a durable response to immunotherapy. Despite these rapid advances in the understanding and management of patients with MCC, many basic, translational and clinical research questions remain unanswered. In March 2018, an International Workshop on Merkel Cell Carcinoma Research was held at the US National Cancer Institute, at which academic, government and industry experts met to identify the highest-priority research questions. Here, we review the biology and treatment of MCC and report the consensus-based recommendations agreed upon during the workshop.


Assuntos
Carcinogênese/genética , Carcinoma de Célula de Merkel/tratamento farmacológico , Carcinoma de Célula de Merkel/genética , Imunoterapia/tendências , Antígeno B7-H1/antagonistas & inibidores , Antígeno B7-H1/imunologia , Carcinoma de Célula de Merkel/imunologia , Carcinoma de Célula de Merkel/virologia , Humanos , Polyomavirus/genética , Polyomavirus/patogenicidade , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Receptor de Morte Celular Programada 1/imunologia
16.
J Clin Virol ; 107: 25-28, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30114678

RESUMO

BACKGROUND: WU polyomavirus (WUPyV) is a relatively new virus associated with respiratory infections. However, its role is unclear in children with severe respiratory failure. OBJECTIVES: We aimed to evaluate the characteristics of severe respiratory failure associated with WUPyV infection in children. STUDY DESIGN: We retrospectively reviewed cases of respiratory tract infection at a tertiary children's hospital in Japan and performed real-time polymerase chain reaction (PCR) for WUPyV using residual extracted nucleic acid samples taken from respiratory tract samples of pediatric patients primarily with respiratory failure. We investigated the clinical characteristics of patients positive for WUPyV and assessed samples positive for WUPyV for other respiratory pathogens using multiplex PCR. RESULTS: WUPyV was detected in 14 of 318 specimens of respiratory tract infections. The median age was 34 months and males were predominant (n = 11, 64%). An underlying disease was found in 11 (79%) patients including five preterm and three immunocompromised patients. The most common clinical diagnosis was pneumonia (n = 13, 93%). The majority of the samples were endotracheal tube aspirates (n = 11, 79%). Other viruses were co-detected in nine (64%) patients, while WUPyV was the only pathogen detected in five patients with a history of admission to the neonatal intensive care unit. These five patients presented with fever and cough, and perihilar infiltrates were detected on chest radiograph in several days. CONCLUSIONS: WUPyV was detected in children with severe respiratory failure independently or concurrently with other pathogens. WUPyV can be a pathogen for children with a history of preterm birth or an underlying disease.


Assuntos
Infecções por Polyomavirus/complicações , Polyomavirus/isolamento & purificação , Insuficiência Respiratória/virologia , Criança , Pré-Escolar , DNA Viral , Feminino , Humanos , Hospedeiro Imunocomprometido , Lactente , Transmissão Vertical de Doença Infecciosa/estatística & dados numéricos , Japão , Masculino , Reação em Cadeia da Polimerase Multiplex , Nasofaringe/virologia , Polyomavirus/genética , Polyomavirus/patogenicidade , Infecções por Polyomavirus/diagnóstico , Infecções Respiratórias/virologia , Estudos Retrospectivos
18.
Arch Virol ; 163(11): 3203-3206, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30097743

RESUMO

A novel polyomavirus (PyVs) comprising 5,422 bp was identified by high-throughput sequencing (HTS) in pooled organs of nutria (Myocastor coypus). The new genome displays the archetypal organization of PyVs, which includes open reading frames for the regulatory proteins small T antigen (sTAg) and large T antigen (LTAg), as well as for the capsid proteins VP1, VP2 and VP3. Based on the International Committee on Taxonomy of Viruses (ICTV) Polyomaviridae Study Group criteria, this genome comprises a new PyVs species for the Alphapolyomavirus genus and is putatively named "Myocastor coypus Polyomavirus 1" . The complete genome sequence of this Myocastor coypus Polyomavirus 1 (McPyV1) isolate is publically available under the GenBank accession no. MH182627.


Assuntos
Infecções por Polyomavirus/veterinária , Polyomavirus/isolamento & purificação , Doenças dos Roedores/virologia , Roedores/virologia , Animais , Antígenos Virais de Tumores/genética , Proteínas do Capsídeo/genética , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , Filogenia , Polyomavirus/classificação , Polyomavirus/genética , Infecções por Polyomavirus/virologia , Ratos
19.
Viruses ; 10(8)2018 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-30126238

RESUMO

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. Through this activity, they are implicated in almost every cellular process investigated to date. Hence, it is not surprising that miRNAs play diverse roles in regulation of viral infections and antiviral responses. Diverse families of DNA and RNA viruses have been shown to take advantage of cellular miRNAs or produce virally encoded miRNAs that alter host or viral gene expression. MiRNA-mediated changes in gene expression have been demonstrated to modulate viral replication, antiviral immune responses, viral latency, and pathogenesis. Interestingly, viruses mediate both canonical and non-canonical interactions with miRNAs to downregulate specific targets or to promote viral genome stability, translation, and/or RNA accumulation. In this review, we focus on recent findings elucidating several key mechanisms employed by diverse virus families, with a focus on miRNAs at the host⁻virus interface during herpesvirus, polyomavirus, retroviruses, pestivirus, and hepacivirus infections.


Assuntos
Regulação Viral da Expressão Gênica , Genoma Viral , Herpesviridae/genética , MicroRNAs/genética , Viroses/genética , Hepacivirus/genética , Hepacivirus/crescimento & desenvolvimento , Hepacivirus/patogenicidade , Herpesviridae/crescimento & desenvolvimento , Herpesviridae/patogenicidade , Humanos , Evasão da Resposta Imune/genética , MicroRNAs/classificação , MicroRNAs/imunologia , Conformação de Ácido Nucleico , Pestivirus/genética , Pestivirus/crescimento & desenvolvimento , Pestivirus/patogenicidade , Polyomavirus/genética , Polyomavirus/crescimento & desenvolvimento , Polyomavirus/patogenicidade , RNA Viral/genética , RNA Viral/imunologia , Retroviridae/genética , Retroviridae/crescimento & desenvolvimento , Retroviridae/patogenicidade , Transdução de Sinais , Viroses/imunologia , Viroses/virologia , Latência Viral/genética , Replicação Viral/genética
20.
Mol Cell Probes ; 40: 13-18, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29883628

RESUMO

In this study, we describe a duplex real-time PCR assay for the simultaneous detection of KIPyV and WUPyV polyomaviruses based on TaqMan probes. This assay detected 500 copies/mL both for KIPyV and WUPyV in 100% of tested positive samples. We assessed this technique on 482 nasopharyngeal aspirate specimens from hospitalized pediatric patients with respiratory symptoms, previously analyzed with commercial multiplex assay for 16 major respiratory viruses. Our assay detected KIPyV genome in 15 out of 482 samples (3.1%) and WUPyV genome in 24 out of 482 samples (4.9%), respectively, and in three samples the coinfection of the two viruses was found. Interestingly, 29 out of 36 of samples with KIPyV and/or WUPyV infection exhibited a co-infection with one or more respiratory viruses confirming that KIPyV and WUPyV were often detected in association to other viral infections. Of note, KIPyV and WUPyV were detected singularly in 4 out of 15 cases and 3 out of 24 cases, respectively, suggesting a possible direct role of these viruses in the respiratory diseases. In conclusion, this method could be taken into account as an alternative technical approach to detect KIPyV and/or WUPyV in respiratory samples for epidemiological and diagnostic analyses.


Assuntos
Nasofaringe/virologia , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Criança , Humanos , Padrões de Referência , Reprodutibilidade dos Testes , Infecções Respiratórias/virologia , Sucção
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