Infection kinetics of human adenovirus serotype 41 in HEK 293 cells

Siqueira-Silva, Joselma; Yeda, Fernanda Perez; Favier, Anne-Laure; Mézin, P.; Silva, Misael Leonardo; Barrella, Karina M.; Mehnert, Dolores Ursula; Fender, Pascal; Hársi, Charlotte Marianna

doi:10.1590/s0074-02762009000500013

Cited by 16 publications

(14 citation statements)

References 47 publications

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“…Mammalian Cells HEK-293 cells (human embryonic kidney cells) were cultured in a monolayer with Eagle's Minimum Essential Medium supplemented with 10% of fetal bovine serum (FBS), both from Cultilab (Emcare, Campinas, Brazil) as previously described [10] .…”

Section: Cell Culturementioning

confidence: 99%

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Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41

Siqueira-Silva

Fenel²,

Gout³

et al. 2011

Intervirology

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Objective: In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have constructed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 penton base. Methods: Recombinant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 penton base were cloned and amplified in Sf9 insect cells. Recombinant dodecahedra were expressed by coinfection of High Five™ cells with both baculoviruses, 72 h post-infection. Cell lysate was centrifuged on sucrose density gradient and the purified recombinant dodecahedra were recovered. Results: Analysis by negative staining electron microscopy demonstrated that chimeric dodecahedra made of the HAdV-3 penton base and decorated with the HAdV-41 SF were successfully generated. Next, recombinant dodecahedra were digested with pepsin and analyzed by Western blot. A ‘site-specific’ proteolysis of the HAdV-41 SF was observed, while the HAdV-3 penton base core was completely digested. Conclusion: These results show that, in vitro, the HAdV-41 SF likely undergoes proteolysis in the gastrointestinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results obtained in the present study may be the basis for the development of gene therapy vectors towards the intestinal epithelium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification.

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Section: Cell Culturementioning

confidence: 99%

“…They are important etiological agents of infantile gastroenteritis [9] and apparently do not cause diseases outside the gastrointestinal tract, which make them attractive as vectors for therapy or vaccination [10] . The molecular basis of their specific pathogenesis is not known, but may be related to the capsid structures.…”

Section: Introductionmentioning

confidence: 99%

Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41

Siqueira-Silva

Fenel²,

Gout³

et al. 2011

Intervirology

Self Cite

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“…HEK 293 cells are very easy to grow and transfect very readily and have been widely used in cell biology research for many years. They are also used by the biotechnology industry to produce therapeutic proteins and viruses for gene therapy [4], [5]. Scrophularia striata which grows in Ilam province is used as a traditional medicine for treatment of diseases for years.…”

Section: Introductionmentioning

confidence: 99%

The Effects of Scrophularia striata Extract on HEK Cell Line in Cell Culture

2016

2016 International Conference on Biological and Environmental Science (BIOES-16) June 8-9, 2016 New Jersey (USA)

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“…In addition, virus detection by PCR methods can be used for specific monitoring of multiple virus types simultaneously, while plaque assays may completely fail to detect certain virus types, when fast growing competitive strains are present [19,20]. For example human adenovirus [hAdV] serotype 2 is released twice as fast by infected HEK293 cells as the slow growing fecal adenoviruses and may completely lyse a cell layer before human adenovirus serotype 41 will show any visible effect, especially when different amounts of these viruses are present in the same sample [21]. Moreover, adenovirus cultivation is clearly influenced by the cell line used.…”

Section: Introductionmentioning

confidence: 99%

Monitoring of adenovirus serotypes in environmental samples by combined PCR and melting point analyses

Hartmann

Dartscht

Szewzyk

et al. 2013

Virol J

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BackgroundHuman adenoviruses are promising candidates for addressing health risks associated with enteric viruses in environmental waters. Relatively harmless but common, these DNA viruses persist within the population and are generally considered extremely stable, remaining infectious in water for long periods of time. Group-specific or single species detection of human adenoviruses in environmental samples is usually based on polymerase chain reaction assays. Simultaneous identification of specific species or serotypes needs additional processing. Here we present a simple molecular approach for the monitoring of serotypic diversity in the human adenovirus populations in contaminated water sites.MethodsDiversity patterns of human adenoviruses in environmental samples, collected in an outdoor artificial stream and pond simulation system, were analyzed using a closed tube polymerase chain reaction method with subsequent melting point analysis.ResultsHuman adenovirus serotype 41 was the most prominent adenovirus serotype detected in environmental water samples, but melting point analyses indicated the presence of additional adenovirus serotypes.ConclusionsBased on investigations with spiked and environmental samples, a combination of qPCR and melting point analysis was shown to identify adenovirus serotypes in sewage contaminated water.

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Infection kinetics of human adenovirus serotype 41 in HEK 293 cells

Abstract: The purpose of this work was to acquire an overview of the infectious cycle of

Cited by 16 publications

References 47 publications

Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41

Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41

The Effects of Scrophularia striata Extract on HEK Cell Line in Cell Culture

Monitoring of adenovirus serotypes in environmental samples by combined PCR and melting point analyses

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