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1.
BMC Infect Dis ; 21(1): 213, 2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33632148

RESUMO

BACKGROUND: Previous studies have demonstrated an association between adenovirus viremia and disease severity in immunocompromised children. However, few studies have focused on this association in immunocompetent children. This study explored the association between adenovirus viremia and adenovirus pneumonia severity in immunocompetent children. METHODS: We performed a retrospective, observational study of immunocompetent children with adenovirus pneumonia admitted to Shenzhen Children's Hospital in Shenzhen, China. Pneumonia was classified as severe or mild based on the Chinese guideline for the classification of pneumonia severity. Serum samples from all the children included in the study were tested for adenovirus DNA with a quantitative polymerase chain reaction. Clinical manifestations, laboratory examinations, and disease severity were compared between children with severe and mild pneumonia. RESULTS: A total of 111 immunocompetent children with adenovirus pneumonia (60 severe, 51 mild) were included. The median age was 40 months, and 64 patients were male. Five patients were admitted to the intensive care unit, and two underwent endotracheal intubation. All patients were discharged after recovery or improvement. Univariate analysis and binary logistic regression analysis showed that leukocytosis (OR = 1.1; 95% CI: 1.0 to 1.2; P = 0.033), co-infection with Mycoplasma pneumoniae (OR = 5.0; 95% CI: 2.1 to 12.3; P <  0.001), and high blood viral load (OR = 1.5; 95% CI: 1.2 to 2.0; P = 0.001) may be risk factors for severe adenovirus pneumonia. CONCLUSIONS: Leukocytosis, co-infection with Mycoplasma pneumoniae, and high blood viral load may be risk factors for severe adenovirus pneumonia in immunocompetent children. Blood viral load may predict pneumonia severity.


Assuntos
Adenoviridae/fisiologia , Infecções por Adenovirus Humanos/virologia , Pneumonia Viral/virologia , Viremia/virologia , Infecções por Adenovirus Humanos/sangue , Infecções por Adenovirus Humanos/epidemiologia , Criança , Pré-Escolar , China/epidemiologia , Feminino , Hospitais Pediátricos , Humanos , Lactente , Masculino , Pneumonia Viral/sangue , Pneumonia Viral/epidemiologia , Estudos Retrospectivos , Fatores de Risco , Índice de Gravidade de Doença , Carga Viral , Viremia/epidemiologia
2.
Int J Mol Sci ; 22(1)2021 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-33466472

RESUMO

A viral infection that involves virus invasion, protein synthesis, and virion assembly is typically accompanied by sharp fluctuations in the intracellular levels of metabolites. Under certain conditions, dramatic metabolic shifts can result in various types of cell death. Here, we review different types of adenovirus-induced cell death associated with changes in metabolic profiles of the infected cells. As evidenced by experimental data, in most cases changes in the metabolome precede cell death rather than represent its consequence. In our previous study, the induction of autophagic cell death was observed following adenovirus-mediated lactate production, acetyl-CoA accumulation, and ATP release, while apoptosis was demonstrated to be modulated by alterations in acetate and asparagine metabolism. On the other hand, adenovirus-induced ROS production and ATP depletion were demonstrated to play a significant role in the process of necrotic cell death. Interestingly, the accumulation of ceramide compounds was found to contribute to the induction of all the three types of cell death mentioned above. Eventually, the characterization of metabolite analysis could help in uncovering the molecular mechanism of adenovirus-mediated cell death induction and contribute to the development of efficacious oncolytic adenoviral vectors.


Assuntos
Adenoviridae/genética , Adenoviridae/fisiologia , Morte Celular/genética , Morte Celular/fisiologia , Metaboloma/genética , Metaboloma/fisiologia , Apoptose/genética , Apoptose/fisiologia , Vetores Genéticos/genética , Vetores Genéticos/fisiologia , Humanos
3.
Ann Hematol ; 100(3): 753-761, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33439306

RESUMO

Adenovirus (ADV)- or BK virus (BKV)-associated hemorrhagic cystitis (HC) is a common complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Several risk factors have been previously reported; however, it is unclear whether virus-associated HC can be transmitted. To clarify this point, we performed a retrospective cohort study on 207 consecutive patients who underwent allo-HSCT at Kyoto University Hospital between 2012 and 2018. We evaluated the incidence and risk factors of virus-associated HC and performed a phylogenetic analysis of the ADV partial sequence. The median age at transplantation was 50 (range, 17-68) years. Fifty-eight patients (28%) developed HC. ADVs were detected in 18 cases, BKVs were detected in 51, both were detected in 12, and only John Cunningham virus (JCV) was detected in 1 case. No factor was significantly associated with HC. However, both ADV- and BKV-HC occurred intensively between April 2016 and September 2017, which suggested possible nosocomial transmission of ADV and BKV. Genome sequencing of the hexon, E3, and penton regions of detected ADVs identified 7 cases of ADV type 11, 2 cases of type 35, and 3 cases of a type 79-related strain. A sequence analysis revealed that these strains in each type were almost identical, except for one case of a type 79-related strain. In conclusion, ADV-HCs with possible nosocomial transmission were described based on genotyping of the virus and partial sequencing of the viral genome. Although viral HC after allo-HSCT is thought to mainly be due to reactivation of a latent virus, nosocomial transmission of ADV or BKV should also be considered.


Assuntos
Infecção Hospitalar/etiologia , Cistite/virologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Hemorragia/virologia , Viroses/etiologia , Adenoviridae/isolamento & purificação , Adenoviridae/fisiologia , Infecções por Adenoviridae/epidemiologia , Infecções por Adenoviridae/etiologia , Adolescente , Adulto , Idoso , Vírus BK/isolamento & purificação , Vírus BK/fisiologia , Estudos de Coortes , Infecção Hospitalar/epidemiologia , Cistite/epidemiologia , Cistite/etiologia , Feminino , Transplante de Células-Tronco Hematopoéticas/estatística & dados numéricos , Hemorragia/epidemiologia , Hemorragia/etiologia , Humanos , Vírus JC/isolamento & purificação , Vírus JC/fisiologia , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/etiologia , Estudos Retrospectivos , Fatores de Risco , Transplante Homólogo/efeitos adversos , Transplante Homólogo/estatística & dados numéricos , Infecções Tumorais por Vírus/epidemiologia , Infecções Tumorais por Vírus/etiologia , Infecções Urinárias/epidemiologia , Infecções Urinárias/etiologia , Viroses/epidemiologia , Adulto Jovem
4.
Anticancer Res ; 41(2): 773-782, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33517282

RESUMO

BACKGROUND/AIM: Oncolytic adenoviruses (OAds) have attracted much attention as novel anticancer therapeutics. The proper design of an expression cassette containing the E1A gene, which is indispensable for self-replication of the Ad genome, is crucial for efficient tumor cell-specific infection of an OAd. Various types of oncolytic adenoviruses (OAds) possessing different types of the E1A gene expression cassettes have been developed, but their oncolytic activities and safety profiles have not been systematically evaluated. Herein we examined the oncolytic activities and safety profiles of five types of OAds possessing different types of the E1A gene expression cassette in order to optimize the E1A gene expression cassette for development of an efficient and safe OAd. MATERIALS AND METHODS: We prepared five types of OAds containing different types of E1 gene expression cassettes, and examined the oncolytic activities and safety profiles of the OAds. RESULTS: Among the OAds examined, OAd-Δ24, which had a 24-bp deletion in the E1A gene, mediated the most efficient oncolytic activities against the human tumor cell lines, although OAd-Δ24 showed slightly higher cytotoxicity to normal human cells than the other OAds. CONCLUSION: These results provide important clues for the development of safe and efficient OAds.


Assuntos
Adenoviridae/fisiologia , Proteínas E1A de Adenovirus/genética , Deleção de Sequência , Survivina/genética , Telomerase/genética , Adenoviridae/genética , Apoptose , Linhagem Celular Tumoral , Expressão Gênica , Células HCT116 , Células HEK293 , Células HeLa , Células Hep G2 , Células Endoteliais da Veia Umbilical Humana , Humanos , Células MCF-7 , Terapia Viral Oncolítica , Vírus Oncolíticos/genética , Vírus Oncolíticos/fisiologia , Regiões Promotoras Genéticas , Replicação Viral
5.
Nat Biomed Eng ; 5(2): 179-189, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33495639

RESUMO

Base editors are RNA-programmable deaminases that enable precise single-base conversions in genomic DNA. However, off-target activity is a concern in the potential use of base editors to treat genetic diseases. Here, we report unbiased analyses of transcriptome-wide and genome-wide off-target modifications effected by cytidine base editors in the liver of mice with phenylketonuria. The intravenous delivery of intein-split cytidine base editors by dual adeno-associated viruses led to the repair of the disease-causing mutation without generating off-target mutations in the RNA and DNA of the hepatocytes. Moreover, the transient expression of a cytidine base editor mRNA and a relevant single-guide RNA intravenously delivered by lipid nanoparticles led to ~21% on-target editing and to the reversal of the disease phenotype; there were also no detectable transcriptome-wide and genome-wide off-target edits. Our findings support the feasibility of therapeutic cytidine base editing to treat genetic liver diseases.


Assuntos
Citidina/genética , DNA/genética , Edição de Genes/métodos , Hepatócitos/metabolismo , RNA/genética , Adenoviridae/fisiologia , Animais , Vetores Genéticos/fisiologia , Células HEK293 , Humanos , Camundongos Endogâmicos C57BL
6.
Int J Nanomedicine ; 15: 6327-6338, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32922004

RESUMO

Purpose: To construct a three-dimensional (3D) culture model of adenovirus in vitro using the nanoself-assembling peptide RADA16-I as a 3D cell culture scaffold combined with virology experimental technology to provide a novel research method for virus isolation and culture, pathogenesis research, antiviral drug screening and vaccine preparation. Methods: The nanoself-assembling peptide RADA16-I was used as a 3D scaffold material for 293T cell culture, and adenovirus was cultured in the cells. The growth, morphological characteristics and pathological effects of 3D-cultured 293T cells after adenovirus infection were observed with an inverted microscope and MTS. The proliferation of adenovirus in 293T cells was observed by TEM and detected by qPCR. The levels of TNF-α and IL-8 secreted by adenovirus-infected 293T cells in the RADA16-I 3D culture system were detected by ELISA. Results: The 293T cells grew well in the RADA16-I 3D culture system for a prolonged period of time. The adenovirus infection persisted for a long time with multiple proliferation peaks, which closely resembled those of in vivo infections. The adenovirus virions amplified in the 3D system remained infectious. There were multiple secretion peaks of TNF-α and IL-8 secretion levels in adenovirus-infected 293T cells cultured in 3D culture systems. Conclusion: The nanoself-assembling peptide RADA16-I can be used as a 3D scaffold for adenovirus isolation, culture and research. The 3D culture system shows more realistic in vivo effects than two-dimensional (2D) culture.


Assuntos
Infecções por Adenoviridae/virologia , Adenoviridae/fisiologia , Técnicas de Cultura de Células/métodos , Nanopartículas/química , Peptídeos/química , Adenoviridae/crescimento & desenvolvimento , Adenoviridae/ultraestrutura , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Células HEK293 , Humanos , Vírion/ultraestrutura
7.
Viruses ; 12(7)2020 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-32630840

RESUMO

Many geometric forms are found in nature, some of them adhering to mathematical laws or amazing aesthetic rules. One of the best-known examples in microbiology is the icosahedral shape of certain viruses with 20 triangular facets and 12 edges. What is less known, however, is that a complementary object displaying 12 faces and 20 edges called a 'dodecahedron' can be produced in huge amounts during certain adenovirus replication cycles. The decahedron was first described more than 50 years ago in the human adenovirus (HAdV3) viral cycle. Later on, the expression of this recombinant scaffold, combined with improvements in cryo-electron microscopy, made it possible to decipher the structural determinants underlying their architecture. Recently, this particle, which mimics viral entry, was used to fish the long elusive adenovirus receptor, desmoglein-2, which serves as a cellular docking for some adenovirus serotypes. This breakthrough enabled the understanding of the physiological role played by the dodecahedral particles, showing that icosahedral and dodecahedral particles live more than a simple platonic story. All these points are developed in this review, and the potential use of the dodecahedron in therapeutic development is discussed.


Assuntos
Adenoviridae/fisiologia , Capsídeo/fisiologia , Infecções por Adenoviridae/patologia , Animais , Proteínas do Capsídeo/fisiologia , Microscopia Crioeletrônica , Humanos , Replicação Viral/fisiologia
8.
PLoS One ; 15(7): e0236175, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32697798

RESUMO

Adenoviruses cause upper respiratory infections, conjunctivitis, keratitis, and gastrointestinal illness. These can be fatal in immunocompromised individuals. Adenoviruses have also been engineered into viral vectors to deliver therapeutic genes or induce immunity as vaccine carriers. The success of ocular gene therapy is driven partly by the immunologic and biochemical influences of the intraocular environment. We have shown that versican and hyaluronan modulate adenoviral vector transgene expression through CD44 signaling. Herein we explored the role of these pathways on virus replication and viral protein expression of wild type adenovirus. We report that the addition of vitreous humor (which contains both versican and hyaluronan) increases viral hexon protein levels. Vitreous humor also increased wild type adenovirus DNA replication in vitro. Metalloproteinase and γ-secretase inhibitors, which inhibit CD44 proteolytic activation, blocked adenoviral replication in vitro. Similarly, protein kinase C and RhoA kinase inhibitors, both proteins associated with CD44 mediated pathways, also inhibited wild type adenoviral replication in vitro. Application of metalloproteinase and γ-secretase inhibitors to human conjunctival explants sharply decreased adenoviral vector gene expression. Our results demonstrate that pharmacologic delivery of these inhibitors is easily achievable. The inhibition of these enzymes should be explored as potential therapies of wild type adenoviral infections.


Assuntos
Infecções por Adenoviridae/tratamento farmacológico , Adenoviridae/efeitos dos fármacos , Antivirais/farmacologia , Vetores Genéticos/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Adenoviridae/fisiologia , Infecções por Adenoviridae/virologia , Administração Oftálmica , Amidas/farmacologia , Amidas/uso terapêutico , Antivirais/uso terapêutico , Túnica Conjuntiva/metabolismo , DNA Viral/genética , DNA Viral/isolamento & purificação , Diaminas/farmacologia , Diaminas/uso terapêutico , Dipeptídeos/farmacologia , Dipeptídeos/uso terapêutico , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Vetores Genéticos/fisiologia , Células HeLa , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/metabolismo , Ácidos Hidroxâmicos/farmacologia , Ácidos Hidroxâmicos/uso terapêutico , Indóis/farmacologia , Indóis/uso terapêutico , Maleimidas/farmacologia , Maleimidas/uso terapêutico , Metaloproteases/antagonistas & inibidores , Metaloproteases/metabolismo , Permeabilidade , Proteína Quinase C/antagonistas & inibidores , Proteína Quinase C/metabolismo , Proteólise/efeitos dos fármacos , Piridinas/farmacologia , Piridinas/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Tiazóis/farmacologia , Tiazóis/uso terapêutico , Versicanas/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo , Corpo Vítreo/metabolismo , Quinases Associadas a rho/metabolismo
9.
J Virol ; 94(10)2020 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-32161167

RESUMO

Nuclear import of viral genomes is an important step during the life cycle of adenoviruses (AdV), requiring soluble cellular factors as well as proteins of the nuclear pore complex (NPC). We addressed the role of the cytoplasmic nucleoporin Nup358 during adenoviral genome delivery by performing depletion/reconstitution experiments and time-resolved quantification of adenoviral genome import. Nup358-depleted cells displayed reduced efficiencies of nuclear import of adenoviral genomes, and the nuclear import receptor transportin 1 became rate limiting under these conditions. Furthermore, we identified a minimal N-terminal region of Nup358 that was sufficient to compensate for the import defect. Our data support a model where Nup358 functions as an assembly platform that promotes the formation of transport complexes, allowing AdV to exploit a physiological protein import pathway for accelerated transport of its DNA.IMPORTANCE Nuclear import of viral genomes is an essential step to initiate productive infection for several nuclear replicating DNA viruses. On the other hand, DNA is not a physiological nuclear import substrate; consequently, viruses have to exploit existing physiological transport routes. Here, we show that adenoviruses use the nucleoporin Nup358 to increase the efficiency of adenoviral genome import. In its absence, genome import efficiency is reduced and the transport receptor transportin 1 becomes rate limiting. We show that the N-terminal half of Nup358 is sufficient to drive genome import and identify a transportin 1 binding region. In our model, adenovirus genome import exploits an existing protein import pathway and Nup358 serves as an assembly platform for transport complexes.


Assuntos
Adenoviridae/genética , Adenoviridae/fisiologia , Chaperonas Moleculares/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , beta Carioferinas/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Genoma Viral , Células HEK293 , Células HeLa , Humanos , Chaperonas Moleculares/química , Poro Nuclear/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/química , Transporte Proteico , Receptores Citoplasmáticos e Nucleares/metabolismo , beta Carioferinas/química
11.
PLoS One ; 15(2): e0228451, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32017805

RESUMO

Viral reactivation occurs frequently in the context of immunodeficiency and immunosuppression after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and can cause severe complications. The aim of this single-center retrospective analysis was to characterize viral infections in the first year after HSCT, to investigate risk factors and to study the impact of viral infections on transplantation outcome. This will facilitate the identification of at-risk patients and the development of new preventive strategies. 107 pediatric allo-HSCT from January 2005 through December 2015 were analyzed for infections with Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpesvirus 6 (HHV-6), adenovirus (ADV), herpes simplex virus (HSV) and varicella zoster virus (VZV). Viral infections were detected after 68.2% of transplantations. The viruses most commonly encountered were HHV-6 (36/107) and EBV (30/107). Severe viral disease was rare (7/107) and none of the patients died as result of viral reactivation. Important risk factors for viral infections were higher age at HSCT, donor type and occurrence of acute graft-versus-host disease (aGvHD). Especially for EBV, transplant from an unrelated donor and in-vivo T-cell depletion (TCD) had a significant effect on infection rates, whereas for CMV the strongest effect was seen by donor and recipient serostatus with recipient seropositivity most predictive for reactivation. The occurrence of severe aGvHD was associated with EBV and ADV infections. For HSV, the recipient serostatus was identified as prognostic factor for HSV infections, while we found higher age at time of HSCT as risk factor for VZV infections. The overall survival of patients with or without viral infections did not differ significantly. Interestingly, when looking at the 85 patients in our cohort who had received an HSCT for a malignant disease, a tendency towards lower relapse rates was seen in patients affected by viral infections (HR 0.51, 95% CI 0.25 - 1.06, p = 0.072). Viral reactivations are common after pediatric allo-HSCT, though severe complications were rare in our collective. Determining risk factors for viral reactivations may help to identify patients in need of intensified monitoring and to individualize preventive strategies.


Assuntos
Doença Enxerto-Hospedeiro/epidemiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Ativação Viral , Viroses/epidemiologia , Adenoviridae/fisiologia , Adolescente , Adulto , Criança , Pré-Escolar , Citomegalovirus/fisiologia , Feminino , Doença Enxerto-Hospedeiro/virologia , Herpesvirus Humano 3/fisiologia , Herpesvirus Humano 4/fisiologia , Herpesvirus Humano 6/fisiologia , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Retrospectivos , Medição de Risco , Simplexvirus/fisiologia , Análise de Sobrevida , Transplante Homólogo/efeitos adversos , Viroses/virologia , Adulto Jovem
12.
Sci Rep ; 10(1): 425, 2020 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-31949228

RESUMO

Mesenchymal stem cells (MSCs) loaded with oncolytic viruses are presently being investigated as a new modality of advanced/metastatic tumors treatment and enhancement of virotherapy. MSCs can, however, either promote or suppress tumor growth. To address the critical question of how MSCs loaded with oncolytic viruses affect virotherapy outcomes and tumor growth patterns in a tumor microenvironment, we developed and analyzed an integrated mathematical-experimental model. We used the model to describe both the growth dynamics in our experiments of firefly luciferase-expressing Hep3B tumor xenografts and the effects of the immune response during the MSCs-based virotherapy. We further employed it to explore the conceptual clinical feasibility, particularly, in evaluating the relative significance of potential immune promotive/suppressive mechanisms induced by MSCs loaded with oncolytic viruses. We were able to delineate conditions which may significantly contribute to the success or failure of MSC-based virotherapy as well as generate new hypotheses. In fact, one of the most impactful outcomes shown by this investigation, not inferred from the experiments alone, was the initially counter-intuitive fact that using tumor-promoting MSCs as carriers is not only helpful but necessary in achieving tumor control. Considering the fact that it is still currently a controversial debate whether MSCs exert a pro- or anti-tumor action, mathematical models such as this one help to quantitatively predict the consequences of using MSCs for delivering virotherapeutic agents in vivo. Taken together, our results show that MSC-mediated systemic delivery of oncolytic viruses is a promising strategy for achieving synergistic anti-tumor efficacy with improved safety profiles.


Assuntos
Adenoviridae/fisiologia , Células-Tronco Mesenquimais/metabolismo , Modelos Biológicos , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/fisiologia , Adenoviridae/metabolismo , Proliferação de Células , Vírus Oncolíticos/metabolismo
13.
Clin Lab ; 65(12)2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31850714

RESUMO

BACKGROUND: Viral gastroenteritis is one of the most common illnesses in humans worldwide, and different viral agents have been shown to be associated with the disease. Among these, rotaviruses and adenoviruses are the responsible causative agents of acute gastroenteritis and causing numerous outbreaks. Therefore, a simple and rapid diagnostic tool, such as an immunochromatographic (IC) test, is required for rapid diagnosis, especially during an outbreak of these pathogens. METHODS: The efficiency of two commercial IC kits were evaluated for simultaneous detections of rotavirus and adenovirus in clinical stool specimens by a single test kit. RESULTS: The data demonstrated that both IC test kits could detect either adenovirus or rotavirus positive alone, as well as mixed infections of both viruses in a single stool specimen. In addition, a wide variety of rotavirus genotypes, including G1-P[8]-I1, G2-P[4]-I2, G3-P[8]-I2, G8-P[8]-I2, and G9-P[8]-I1 could be detected by both IC kits. The detection limit of the kits for the detection of rotavirus and adenovirus were comparable to those of real-time PCR at 105 copies/mL. CONCLUSIONS: These two IC test kits could be used as an alternative choice for rapid screening of rotavirus and adenovirus in the stool specimens, especially during the seasonal outbreak of acute gastroenteritis.


Assuntos
Infecções por Adenoviridae/diagnóstico , Adenoviridae/genética , Cromatografia/métodos , Imunoensaio/métodos , Infecções por Rotavirus/diagnóstico , Rotavirus/genética , Adenoviridae/fisiologia , Infecções por Adenoviridae/complicações , Infecções por Adenoviridae/virologia , Pré-Escolar , Fezes/virologia , Gastroenterite/complicações , Gastroenterite/diagnóstico , Genótipo , Humanos , Lactente , Kit de Reagentes para Diagnóstico/normas , Reprodutibilidade dos Testes , Rotavirus/fisiologia , Infecções por Rotavirus/complicações , Infecções por Rotavirus/virologia , Sensibilidade e Especificidade
14.
Cold Spring Harb Protoc ; 2019(12)2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31792138

RESUMO

The sensitivity of an assay for replication-competent adenoviruses (RCAs) can often be enhanced by biological amplification of the RCAs with serial passage. Here, we describe an extension of this technique, termed "concentration passage," in which RCA replicated during the first plating of the vector is collected and concentrated onto one-tenth of the original number of cells. This significantly increases the chances of detecting the RCAs. Combining this approach with the use of quantitative polymerase chain reaction (qPCR) for sensitive detection of the RCA E1 gene, we are able to reach levels of sensitivity of 1 IU of RCAs in 1011 vector particles. The protocol described here is tailored for HuAd5 vectors using wild-type HuAd5 as the RCA surrogate. However, we have also adapted this technique with similar sensitivity to vectors based on other adenovirus serotypes. If other adenovirus serotypes are assayed, careful consideration should be given to the appropriate RCA surrogate. Strictly speaking, if the vector is propagated in HEK-293 or similar cell lines, the RCA surrogate should be a hybrid virus containing the HuAd5 E1 gene.


Assuntos
Adenoviridae/genética , Adenoviridae/fisiologia , Bioensaio/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Replicação Viral , Células A549 , Análise de Dados , Coleta de Dados , Humanos
15.
FEBS Lett ; 593(24): 3518-3530, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31710378

RESUMO

Adenoviruses induce an extensive reorganization of the host cell nucleus during replication. Such a process results in the assembly of viral and cellular macromolecules into nuclear structures called adenovirus replication compartments (AdRCs), which function as platforms for viral DNA replication and gene expression. AdRCs co-opt host proteins and cellular pathways that restrict viral replication, suggesting that the mechanisms that control AdRC formation and function are essential for viral replication and lay at the basis of virus-host interactions. Here, we review the hallmarks of AdRCs and recent progress in our understanding of the formation, composition, and function of AdRCs. Furthermore, we discuss how AdRCs facilitate the interplay between viral and cellular machineries and hijack cellular functions to promote viral genome replication and expression.


Assuntos
Infecções por Adenoviridae/virologia , Adenoviridae/fisiologia , DNA Viral/genética , Adenoviridae/genética , Infecções por Adenoviridae/metabolismo , Animais , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Regulação Viral da Expressão Gênica , Interações Hospedeiro-Patógeno , Humanos , Proteínas Virais/genética , Replicação Viral
16.
FEBS Lett ; 593(24): 3660-3673, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31747467

RESUMO

Adenoviruses (AdVs) infect representatives of numerous species from almost every major vertebrate class, albeit their incidence shows great variability. AdVs infecting birds, reptiles, and bats are the most common and diverse, whereas only one AdV has been so far isolated both from fish and amphibians. The family Adenoviridae is divided into five genera, each corresponding to an independent evolutionary lineage that supposedly coevolved with its respective vertebrate hosts. Members of genera Mastadenovirus and Aviadenovirus seem to infect exclusively mammals and birds, respectively. The genus Ichtadenovirus includes the single known AdV from fish. The majority of AdVs in the genus Atadenovirus originated from squamate reptiles (lizards and snakes), but also certain mammalian and avian AdVs are classified within this genus. The genus Siadenovirus contains the only AdV isolated from frog, along with numerous avian AdVs. In turtles, members of a sixth AdV lineage have been discovered, pending official recognition as an independent genus. The most likely scenario for AdV evolution includes long-term cospeciation with the hosts, as well as occasional switches between closely or, rarely, more distantly related hosts.


Assuntos
Infecções por Adenoviridae/virologia , Adenoviridae/classificação , Adenoviridae/fisiologia , Infecções por Adenoviridae/veterinária , Animais , Evolução Molecular , Especificidade de Hospedeiro , Filogenia
17.
FEBS Lett ; 593(24): 3395-3418, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31777951

RESUMO

Animal viruses are well recognized for their ability to uncover fundamental cell and molecular processes, and adenovirus certainly provides a prime example. This review illustrates the lessons learned from studying adenovirus over the past five decades. We take a look back at the key studies of adenovirus structure and biophysical properties, which revealed the mechanisms of adenovirus association with antibody, cell receptor, and immune molecules that regulate infection. In addition, we discuss the critical contribution of studies of adenovirus gene expression to elucidation of fundamental reactions in pre-mRNA processing and its regulation. Other pioneering studies furnished the first examples of protein-primed initiation of DNA synthesis and viral small RNAs. As a nonenveloped virus, adenoviruses have furnished insights into the modes of virus attachment, entry, and penetration of host cells, and we discuss the diversity of cell receptors that support these processes, as well as membrane penetration. As a result of these extensive studies, adenovirus vectors were among the first to be developed for therapeutic applications. We highlight some of the early (unsuccessful) trials and the lessons learned from them.


Assuntos
Adenoviridae/fisiologia , Vetores Genéticos/administração & dosagem , Receptores de Superfície Celular/metabolismo , Adenoviridae/genética , Animais , Ensaios Clínicos como Assunto , Vetores Genéticos/uso terapêutico , Humanos , Ligação Viral , Internalização do Vírus
18.
Emerg Microbes Infect ; 8(1): 1679-1687, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31749409

RESUMO

Adenoviruses (AdVs) are major contributors to clinical illnesses. Novel human and animal AdVs continue to be identified and characterized. Comparative analyses using bioinformatic methods and Omics-based technologies allow insights into how these human pathogens have emerged and their potential for host cross-species transmission. Systematic review of literature published across ProQuest, Pubmed, and Web of Science databases for evidence of adenoviral zoonotic potential identified 589 citations. After removing duplicates, 327 citations were screened for relevance; of which, 74 articles received full-text reviews. Among these, 24 were included here, of which 16 demonstrated evidence of zoonotic transmission of AdVs. These documented instances of AdV crossing host species barriers between humans and non-human primate, bat, feline, swine, canine, ovine, and caprine. Eight studies sought to but did not find evidence of zoonosis. The findings demonstrate substantial evidence suggesting AdVs have previously and will continue crossing host species barriers. These have human health consequences both in terms of novel pathogen emergence and epidemic outbreaks, and of appropriate and safe use of non-human adenoviruses for therapeutics. As routine human clinical diagnostics may miss a novel cross-species adenovirus infection in humans, next generation sequencing or panspecies molecular diagnostics may be necessary to detect such incursions.


Assuntos
Infecções por Adenoviridae/transmissão , Infecções por Adenoviridae/veterinária , Adenoviridae/fisiologia , Zoonoses/transmissão , Adenoviridae/genética , Infecções por Adenoviridae/virologia , Animais , Gatos , Cães , Cabras , Especificidade de Hospedeiro , Humanos , Filogenia , Ovinos , Suínos , Zoonoses/virologia
19.
FEBS Lett ; 593(24): 3419-3448, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31758703

RESUMO

Incoming adenoviruses seize control of cytosolic transport mechanisms to relocate their genome from the cell periphery to specialized sites in the nucleoplasm. The nucleus is the site for viral gene expression, genome replication, and the production of progeny for the next round of infection. By taking control of the cell, adenoviruses also suppress cell-autonomous immunity responses. To succeed in their production cycle, adenoviruses rely on well-coordinated steps, facilitated by interactions between viral proteins and cellular factors. Interactions between virus and host can impose remarkable morphological changes in the infected cell. Imaging adenoviruses has tremendously influenced how we delineate individual steps in the viral life cycle, because it allowed the development of specific optical markers to label these morphological changes in space and time. As technology advances, innovative imaging techniques and novel tools for specimen labeling keep uncovering previously unseen facets of adenovirus biology emphasizing why imaging adenoviruses is as attractive today as it was in the past. This review will summarize past achievements and present developments in adenovirus imaging centered on fluorescence microscopy approaches.


Assuntos
Infecções por Adenoviridae/virologia , Adenoviridae/patogenicidade , Adenoviridae/fisiologia , Núcleo Celular/metabolismo , Núcleo Celular/virologia , Microscopia Crioeletrônica , Interações Hospedeiro-Patógeno , Humanos , Microscopia de Força Atômica , Proteínas Virais/metabolismo , Replicação Viral
20.
FEBS Lett ; 593(24): 3484-3495, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31721176

RESUMO

Viruses alter host cell processes to optimize their replication cycle. Human adenoviruses (Ad) encode proteins that promote viral macromolecular synthesis and counteract innate and adaptive responses to infection. The focus of this review is on how Ad evades innate cellular responses to infection, including an interferon (IFN) response and a DNA damage response (DDR). Ad blocks the IFN response by inhibiting cytoplasmic signaling pathways and the activation of IFN-stimulated genes (ISGs), as well as the functions of ISG products, such as PML. Ad also inhibits DDR sensors, for instance, the Mre11-Rad50-Nbs1 complex, and DDR effectors like DNA ligase IV. These innate cellular responses impact many different viruses, and studies on Ad have provided broad insight into these areas.


Assuntos
Infecções por Adenoviridae/imunologia , Adenoviridae/fisiologia , Interferons/metabolismo , Proteínas Virais/metabolismo , Adenoviridae/imunologia , Animais , Redes Reguladoras de Genes , Interações Hospedeiro-Patógeno , Humanos , Evasão da Resposta Imune , Imunidade Inata , Transdução de Sinais
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