Unusual splice sites in the E1A?E1B cotranscripts synthesized in adenovirus type 40-infected A549 cells

1994 ◽  
Vol 139 (3-4) ◽  
pp. 389-402 ◽  
Author(s):  
S. Ishida ◽  
Y. Fujinaga ◽  
K. Fujinaga ◽  
N. Sakamoto ◽  
S. Hashimoto
Keyword(s):  
A549 Cells ◽  
Adenovirus Type ◽  
Splice Sites

scholarly journals iTRAQ-Based and Label-Free Proteomics Approaches for Studies of Human Adenovirus Infections

2013 ◽  
Vol 2013 ◽  
pp. 1-16 ◽  
Author(s):  
Hung V. Trinh ◽  
Jonas Grossmann ◽  
Peter Gehrig ◽  
Bernd Roschitzki ◽  
Ralph Schlapbach ◽  
...  
Keyword(s):  
A549 Cells ◽  
Correlation Coefficients ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Label Free ◽  
Absolute Quantitation ◽  
Software Packages ◽  
Itraq Label ◽  
Isobaric Tags ◽  
Type 3

Both isobaric tags for relative and absolute quantitation (iTRAQ) and label-free methods are widely used for quantitative proteomics. Here, we provide a detailed evaluation of these proteomics approaches based on large datasets from biological samples. iTRAQ-label-based and label-free quantitations were compared using protein lysate samples from noninfected human lung epithelial A549 cells and from cells infected for 24 h with human adenovirus type 3 or type 5. Either iTRAQ-label-based or label-free methods were used, and the resulting samples were analyzed by liquid chromatography (LC) and tandem mass spectrometry (MS/MS). To reduce a possible bias from quantitation software, we applied several software packages for each procedure. ProteinPilot and Scaffold Q+ software were used for iTRAQ-labeled samples, while Progenesis LC-MS and ProgenesisF-T2PQ/T3PQ were employed for label-free analyses. R2 correlation coefficients correlated well between two software packages applied to the same datasets with values between 0.48 and 0.78 for iTRAQ-label-based quantitations and 0.5 and 0.86 for label-free quantitations. Analyses of label-free samples showed higher levels of protein up- or downregulation in comparison to iTRAQ-labeled samples. The concentration differences were further evaluated by Western blotting for four downregulated proteins. These data suggested that the label-free method was more accurate than the iTRAQ method.

scholarly journals Adenovirus Types 11p and 35p Show High Binding Efficiencies for Committed Hematopoietic Cell Lines and Are Infective to These Cell Lines

2000 ◽  
Vol 74 (3) ◽  
pp. 1457-1467 ◽  
Author(s):  
Anna Segerman ◽  
Ya-Fang Mei ◽  
Göran Wadell
Keyword(s):  
Cell Lines ◽  
Hematopoietic Cells ◽  
A549 Cells ◽  
Adenovirus Vector ◽  
Adenovirus Type ◽  
Hematopoietic Cell ◽  
Jurkat Cells ◽  
High Binding ◽  
Hematopoietic Cell Lines

ABSTRACT Hematopoietic cells are attractive targets for gene therapy. However, no satisfactory vectors are currently available. A major problem with the most commonly used adenovirus vectors, based on adenovirus type 2 (Ad2) or Ad5, is their low binding efficiency for hematopoietic cells. In this study we identify two adenovirus serotypes with high affinity for hematopoietic cells. The binding efficiency of prototype serotypes Ad4p, Ad11p, and Ad35p for different committed hematopoietic cell lines representing T cells (Jurkat), B cells (DG75), monocytes (U937-2), myeloblasts (K562), and granulocytes (HL-60) was evaluated and compared to that of Ad5v, the commonly used adenovirus vector, using flow cytometry. In contrast to Ad5v, which bound to less than 10% of the cells in all experiments, Ad11p and Ad35p showed high binding efficiency for all of the different hematopoietic cell lines. Ad4p bound to the lymphocytic cell lines to some extent but less well to the myelomonocytic cell lines. The abilities of the different serotypes to infect, replicate, and form complete infectious particles in the hematopoietic cell lines were also investigated by immunostaining, 35S labeling of viral proteins, and titrations of cell lysates. Ad11p and Ad35p infected the highest proportion of cells, and Ad11p infected all of the cell lines investigated. The Ad11p hexon was expressed equally well in K562 and A549 cells. Jurkat cells also showed high levels of expression of Ad11p hexons, but the production of infectious particles was low. The binding properties of virions were correlated to their ability to infect and be expressed.

scholarly journals Replication Properties of Human Adenovirus In Vivo and in Cultures of Primary Cells from Different Animal Species

2006 ◽  
Vol 80 (7) ◽  
pp. 3549-3558 ◽  
Author(s):  
Christian Jogler ◽  
Dennis Hoffmann ◽  
Dirk Theegarten ◽  
Thomas Grunwald ◽  
Klaus Überla ◽  
...  
Keyword(s):  
Viral Load ◽  
Animal Species ◽  
A549 Cells ◽  
Human Adenovirus ◽  
Fold Increase ◽  
Adenovirus Type ◽  
Adenoviral Vectors ◽  
Treatment Modalities ◽  
Late Gene Expression

ABSTRACT Oncolytic adenoviruses have emerged as a promising approach for the treatment of tumors resistant to other treatment modalities. However, preclinical safety studies are hampered by the lack of a permissive nonhuman host. Screening of a panel of primary cell cultures from seven different animal species revealed that porcine cells support productive replication of human adenovirus type 5 (Ad5) nearly as efficiently as human A549 cells, while release of infectious virus by cells from other animal species tested was diminished by several orders of magnitude. Restriction of productive Ad5 replication in rodent and rabbit cells seems to act primarily at a postentry step. Replication efficiency of adenoviral vectors harboring different E1 deletions or mutations in porcine cells was similar to that in A549 cells. Side-by-side comparison of the viral load kinetics in blood of swine and mice injected with Ad5 or a replication-deficient adenoviral vector failed to provide clear evidence for virus replication in mice. In contrast, evidence suggests that adenovirus replication occurs in swine, since adenoviral late gene expression produced a 13.5-fold increase in viral load in an individual swine from day 3 to day 7 and 100-fold increase in viral DNA levels in the Ad5-infected swine compared to the animal receiving a replication-deficient adenovirus. Lung histology of Ad5-infected swine revealed a severe interstitial pneumonia. Although the results in swine are based on a small number of animals and need to be confirmed, our data strongly suggest that infection of swine with human adenovirus or oncolytic adenoviral vectors is a more appropriate animal model to study adenoviral pathogenicity or pharmacodynamic and toxicity profiles of adenoviral vectors than infection of mice.

Quantitative proteomic analysis of A549 cells infected with human adenovirus type 2

2019 ◽  
Vol 91 (7) ◽  
pp. 1239-1249 ◽  
Author(s):  
Kareem R. Badr ◽  
Juliana A. Parente‐Rocha ◽  
Lilian C. Baeza ◽  
Fabiola S. Ficcadori ◽  
Menira Souza ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
A549 Cells ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Quantitative Proteomic Analysis ◽  
Adenovirus Type 2

scholarly journals Interaction of Adenovirus Type 5 Fiber with the Coxsackievirus and Adenovirus Receptor Activates Inflammatory Response in Human Respiratory Cells

2006 ◽  
Vol 80 (22) ◽  
pp. 11241-11254 ◽  
Author(s):  
Anna Tamanini ◽  
Elena Nicolis ◽  
Alberto Bonizzato ◽  
Valentino Bezzerri ◽  
Paola Melotti ◽  
...  
Keyword(s):  
Heparan Sulfate ◽  
Nuclear Translocation ◽  
A549 Cells ◽  
Adenovirus Type ◽  
Nuclear Factor Κb ◽  
Mitogen Activated Protein Kinase ◽  
Penton Base ◽  
Respiratory Cells ◽  
Inducible Protein ◽  
Adenovirus Receptor

ABSTRACT The innate immune response to adenovirus (Ad)-derived gene transfer vectors has been shown to initiate immediately after interaction of Ad with respiratory epithelial cells, through the induction of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and JNK mitogen-activated protein kinase (MAPK), nuclear factor κB (NF-κB), and different proinflammatory genes. Ad serotypes 2 or 5 (Ad2/5) enter respiratory epithelia after initial binding of fiber with the coxsackievirus-adenovirus receptor (CAR) or, alternatively, with cell surface heparan sulfate glycosaminoglycans. Ad2/5 internalization is triggered by binding of penton base to cellular RGD-binding integrins. Here we investigated the role of the Ad5 surface domain proteins constituting the vector capsid, namely, the fiber, the penton base, and the hexon, on the transmembrane signals leading to the transcription of the different proinflammatory genes in the human respiratory A549 cell line. Interaction of Ad fiber with CAR activates both ERK1/2 and JNK MAPK and the nuclear translocation of NF-κB, whereas no activation was observed after exposing A549 cells to penton base and hexon proteins. Moreover, interaction of Ad fiber with CAR, but not heparan sulfate proteoglycans, promotes transcription of the chemokines interleukin-8, GRO-α, GRO-γ, RANTES, and interferon-inducible protein 10. These results identify the binding of Ad5 fiber with the cellular CAR as a key proinflammatory activation event in epithelial respiratory cells that is independent of the transcription of Ad5 genes.

scholarly journals Adenovirus Type 37 Uses Sialic Acid as a Cellular Receptor

2000 ◽  
Vol 74 (1) ◽  
pp. 42-48 ◽  
Author(s):  
Niklas Arnberg ◽  
Karin Edlund ◽  
Alistair H. Kidd ◽  
Göran Wadell
Keyword(s):  
Sialic Acid ◽  
Cho Cells ◽  
A549 Cells ◽  
Adenovirus Type ◽  
Class I Mhc ◽  
Mhc I ◽  
Neuraminidase Treatment ◽  
Cellular Attachment ◽  
Protease Treatment ◽  
Adenovirus Receptor

ABSTRACT Two cellular receptors for adenovirus, coxsackievirus-adenovirus receptor (CAR) and major histocompatibility complex class I (MHC-I) α2, have recently been identified. In the absence of CAR, MHC-I α2 has been suggested to serve as a cellular attachment protein for subgenus C adenoviruses, while members from all subgenera except subgenus B have been shown to interact with CAR. We have found that adenovirus type 37 (Ad37) attachment to CAR-expressing CHO cells was no better than that to CHO cells lacking CAR expression, suggesting that CAR is not used by Ad37 during attachment. Instead, we have identified sialic acid as a third adenovirus receptor moiety. First, Ad37 attachment to both CAR-expresing CHO cells and MHC-I α2-expressing Daudi cells was sensitive to neuraminidase treatment, which eliminates sialic acid on the cell surface. Second, Ad37 attachment to sialic acid-expressing Pro-5 cells was more than 10-fold stronger than that to the Pro-5 subline Lec2, which is deficient in sialic acid expression. Third, neuraminidase treatment of A549 cells caused a 60% decrease in Ad37 replication in a fluorescent-focus assay. Moreover, the receptor sialoconjugate is most probably a glycoprotein rather than a ganglioside, since Ad37 attachment to sialic acid-expressing Pro-5 cells was sensitive to protease treatment. Ad37 attachment to Pro-5 cells occurs via α(2→3)-linked sialic acid saccharides rather than α(2→6)-linked ones, since (i) α(2→3)-specific but not α(2→6)-specific lectins blocked Ad37 attachment to Pro-5 cells and (ii) pretreatment of Pro-5 cells with α(2→3)-specific neuraminidase resulted in decreased Ad37 binding. Taken together, these results suggest that, unlike Ad5, Ad37 makes use of α(2→3)-linked sialic acid saccharides on glycoproteins for entry instead of using CAR or MHC-I α2.

scholarly journals CD46 Is a Cellular Receptor for All Species B Adenoviruses except Types 3 and 7

2005 ◽  
Vol 79 (22) ◽  
pp. 14429-14436 ◽  
Author(s):  
Marko Marttila ◽  
David Persson ◽  
Dan Gustafsson ◽  
M. Kathryn Liszewski ◽  
John P. Atkinson ◽  
...  
Keyword(s):  
Chinese Hamster Ovary Cells ◽  
Rabbit Antiserum ◽  
Chinese Hamster ◽  
A549 Cells ◽  
Adenovirus Type ◽  
Host Cells ◽  
Cellular Receptor ◽  
Coxsackie Adenovirus Receptor ◽  
Tract Infections ◽  
Adenovirus Receptor

ABSTRACT The 51 human adenovirus serotypes are divided into six species (A to F). Adenovirus serotypes from all species except species B utilize the coxsackie-adenovirus receptor for attachment to host cells in vitro. Species B adenoviruses primarily cause ocular and respiratory tract infections, but certain serotypes are also associated with renal disease. We have previously demonstrated that adenovirus type 11 (species B) uses CD46 (membrane cofactor protein) as a cellular receptor instead of the coxsackie-adenovirus receptor (A. Segerman et al., J. Virol. 77:9183-9191, 2003). In the present study, we found that transfection with human CD46 cDNA rendered poorly permissive Chinese hamster ovary cells more permissive to infection by all species B adenovirus serotypes except adenovirus types 3 and 7. Moreover, rabbit antiserum against human CD46 blocked or efficiently inhibited all species B serotypes except adenovirus types 3 and 7 from infecting human A549 cells. We also sequenced the gene encoding the fiber protein of adenovirus type 50 (species B) and compared it with the corresponding amino acid sequences from selected serotypes, including all other serotypes of species B. From the results obtained, we conclude that CD46 is a major cellular receptor on A549 cells for all species B adenoviruses except types 3 and 7.

scholarly journals Adenovirus Type 37 Uses Sialic Acid as a Cellular Receptor on Chang C Cells

2002 ◽  
Vol 76 (17) ◽  
pp. 8834-8841 ◽  
Author(s):  
Niklas Arnberg ◽  
Patricia Pring-Åkerblom ◽  
Göran Wadell
Keyword(s):  
Sialic Acid ◽  
Cell Line ◽  
Cell Lines ◽  
A549 Cells ◽  
Adenovirus Type ◽  
Culture Collection ◽  
Cellular Receptor ◽  
Dependent Manner ◽  
C Cells ◽  
Epidemic Keratoconjunctivitis

ABSTRACT Epidemic keratoconjunctivitis (EKC) is a severe eye infection caused mainly by adenovirus type 8 (Ad8), Ad19, and Ad37. We have shown that the EKC-causing adenoviruses use sialic acid as a cellular receptor on A549 cells instead of the coxsackie-adenovirus receptor, which is used by most adenoviruses. Recently, Wu et al. (Virology 279:78-89, 2001) proposed that Ad37 uses a 50-kDa protein as a receptor on Chang C conjunctival cells and that this interaction is independent of sialic acid. According to the American Type Culture Collection, this cell line carries HeLa cell markers and should be considered to be a genital cell line. This prompted us to investigate the function of sialic acid as a cellular receptor for Ad37 in Chang C cells. In this study, we demonstrate that enzymatic removal or lectin-mediated blocking of cell surface sialic acid inhibits the binding of Ad37 virions to Chang C cells, as does soluble, virion-interacting sialic acid-containing substances. The binding was Ca2+ or Mg2+ ion independent and mediated by the knob domain of the trimeric viral fiber polypeptide. Moreover, Ad37 virions infected Chang C cells and two other genital cell lines (HeLa and SiHa) as well as a corneal cell line in a strictly sialic acid-dependent manner. From these results, we conclude that Ad37 uses sialic acid as a major receptor in cell lines derived from both genital and corneal tissues.

scholarly journals A Single-Cycle Adenovirus Type 7 Vaccine for Prevention of Acute Respiratory Disease

Viruses ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 413
Author(s):  
Brianna L. Bullard ◽  
Brigette N. Corder ◽  
Eric A. Weaver
Keyword(s):  
Respiratory Disease ◽  
A549 Cells ◽  
Adenovirus Type ◽  
Wild Type Virus ◽  
Structural Protein ◽  
Acute Respiratory Disease ◽  
Single Cycle ◽  
Replication Competent Virus ◽  
Military Recruits

Adenovirus type 7 (Ad7) infection is associated with acute respiratory disease (ARD), especially in military recruits living in close quarters. Recently, several outbreaks of Ad7 infections have occurred in civilian populations, with some cases leading to death. However, the current Ad7 vaccine is licensed for use only in military recruits because it utilizes an orally delivered wild type virus which is shed in the stool for 28 days after immunization. This poses a safety risk due to the possibility of virus spread to vulnerable populations. To address the need for a safer Ad7 vaccine for use in civilian populations, we developed a single-cycle Ad7 virus (scAd7). This scAd7 virus is deleted for the Ad7 fiber protein, so that viruses produced outside of complementing cells lines lack this essential structural protein and have severely reduced infectivity. In vitro studies in noncomplementing A549 cells showed that the scAd7 virus has genomic DNA replication kinetics and Ad7 hexon expression similar to a replication-competent virus; however, virus progeny produced after infection has impaired infectivity. Therefore, this scAd7 virus combines the safety advantages of a replication-defective virus with the increased Ad7 gene expression of a replication-competent virus. Due to these advantages, we believe that scAd7 viruses should be further studied as an alternative, safer Adenovirus 7 vaccine.

scholarly journals Rapid Construction of a Replication-Competent Infectious Clone of Human Adenovirus Type 14 by Gibson Assembly

Viruses ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 568 ◽  
Author(s):  
Haibin Pan ◽  
Yuqian Yan ◽  
Jing Zhang ◽  
Shan Zhao ◽  
Liqiang Feng ◽  
...  
Keyword(s):  
Infectious Clone ◽  
A549 Cells ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Enzyme Analysis ◽  
Gibson Assembly ◽  
Assembly Method ◽  
Rapid Construction ◽  
One Step ◽  
Genomic Variant

In 1955, Human adenovirus type 14 (HAdV-B14p) was firstly identified in a military trainee diagnosed as acute respiratory disease (ARD) in the Netherlands. Fifty years later, a genomic variant, HAdV-B14p1, re-emerged in the U.S. and caused large and fatal ARD outbreaks. Subsequently, more and more ARD outbreaks occurred in Canada, the UK, Ireland, and China, in both military and civil settings. To generate a tool for the efficient characterization of this new genomic variant, a full-length infectious genomic clone of HAdV-B14 was successfully constructed using one-step Gibson Assembly method in this study. Firstly, the full genome of HAdV-B14p1 strain GZ01, the first HAdV-B14 isolate in China, was assembled into pBR322 plasmid by Gibson Assembly. The pBRAdV14 plasmid, generated by Gibson Assembly, was analyzed and verified by PCR, restriction enzymes digestion and the sequencing. Secondly, viruses were rescued from pBRAdV14-transfected A549 cells. The integrity of the rescued viruses was identified by restriction enzyme analysis. The complete sequence of the infectious clone was further sequenced. No mutation was found in the infectious clone during the construction when compared with the parental virus and pBR322 sequences. The direct immunofluorescence assay indicated the expression of the hexon protein. Finally, typical virions were observed; the one-step growth curves further showed that the DNA replication and viral reproduction efficiency of pBRAd14 derived viruses was similar with that of wild-type HAdV-B14 strain. The successful construction of the replication-competent infectious clone of pBRAdV14 facilitates the development of vaccine and antiviral drugs against HAdV-B14, as well as provides a novel strategy for rapid construction of infectious viral clones for other large-genome DNA viruses.

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