- MeSH
- adenovirové infekce lidí diagnóza komplikace přenos MeSH
- Caliciviridae patogenita účinky léků MeSH
- dítě MeSH
- dospělí MeSH
- gastroenteritida * komplikace mortalita prevence a kontrola virologie MeSH
- infekce viry z čeledi Astroviridae diagnóza komplikace MeSH
- kultivace virů metody MeSH
- lidé MeSH
- rotavirové infekce komplikace mortalita prevence a kontrola MeSH
- tekutinová terapie metody MeSH
- vakcína proti rotavirům MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
Only very few comparative studies have been performed that evaluate general trends of virus growth under 3D in comparison with 2D cell culture conditions. The aim of this study was to investigate differences when four animal viruses are cultured in 2D and 3D. Suid herpesvirus 1 (SuHV-1), Vesicular stomatitis virus (VSIV), Bovine adenovirus (BAdV) and Bovine parainfluenza 3 virus (BPIV-3) were cultivated in 3D rotating wall vessels (RWVs) and conventional 2D cultures. The production of virus particles, the portion of infectious particles, and the infectious growth curves were compared. For all viruses, the production of virus particles (related to cell density), including the non-infectious ones, was lower in 3D than in 2D culture. The production of only infectious particles was significantly lower in BAdV and BPIV-3 in 3D cultures in relation to cell density. The two cultivation approaches resulted in significantly different virus particle-to-TCID50 ratios in three of the four viruses: lower in SuHV-1 and BPIV-3 and higher in BAdV in 3D culture. The infectious virus growth rates were not significantly different in all viruses. Although 3D RWV culture resulted in lower production of virus particles compared to 2D systems, the portion of infectious particles was higher for some viruses.
- MeSH
- Atadenovirus růst a vývoj fyziologie ultrastruktura MeSH
- buněčné kultury * přístrojové vybavení metody MeSH
- buňky MDCK MeSH
- Cercopithecus aethiops MeSH
- kultivace virů metody MeSH
- prasata MeSH
- prasečí herpesvirus 1 růst a vývoj fyziologie ultrastruktura MeSH
- psi MeSH
- replikace viru MeSH
- skot MeSH
- Vero buňky MeSH
- virus bovinní parainfluenzy 3 růst a vývoj fyziologie ultrastruktura MeSH
- virus vezikulární stomatitidy, kmen Indiana růst a vývoj fyziologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Mouse polyomavirus (MPyV) is a member of the Polyomaviridae family, which comprises non-enveloped tumorigenic viruses infecting various vertebrates including humans and causing different pathogenic responses in the infected organisms. Despite the variations in host tropism and pathogenicity, the structure of the virions of these viruses is similar. The capsid, with icosahedral symmetry (ø, 45 nm, T = 7d), is composed of a shell of 72 capsomeres of structural proteins, arranged around the nucleocore containing approximately 5-kbp-long circular dsDNA in complex with cellular histones. MPyV has been one of the most studied polyomaviruses and serves as a model virus for studies of the mechanisms of cell transformation and virus trafficking, and for use in nanotechnology. It can be propagated in primary mouse cells (e.g., in whole mouse embryo cells) or in mouse epithelial or fibroblast cell lines. In this unit, propagation, purification, quantification, and storage of MPyV virions are presented.
Mumps vaccines are live attenuated viruses. They are known to vary in effectiveness, degree of attenuation and adverse event profile. However, the underlying reasons are poorly understood. We studied two closely related mumps vaccines which originate from the same attenuated Jeryl Lynn-5 strain but have different efficacies. Jeryl Lynn-Canine Kidney (JL-CK), produced on primary canine kidney cells, is less effective than RIT4385, which is produced on chicken embryo fibroblasts. JL-CK and RIT4385 could be distinguished by a number of in vitro and in vivo properties. JL-CK produced heterogeneous, generally smaller plaques than RIT4385, but gave 100-fold higher titres when grown in cells and showed a higher degree of hydrocephalus formation in neonatal rat brains. Sanger sequencing of JL-CK identified 14 regions of heterogeneity throughout the genome. Plaque purification of JL-CK demonstrated the presence of five different Jeryl Lynn-5 variants encompassing the 14 mutations. One JL-CK mutation was associated with a small plaque phenotype, the effects of the others in vitro or in vivo were less clear. Only 4% of the JL-CK population corresponded to the parental Jeryl Lynn-5 strain. Next generation sequencing of JL-CK and virus before and after growth in cell lines or neonatal rat brains showed that propagation in vitro or in vivo altered the population dramatically. Our findings indicate that growth of JL-CK in primary canine kidney cells resulted in the selection of a mixture of mumps virus variants that have different biological properties compared to the parent Jeryl Lynn-5 virus. We also report three previously unknown heterogenic regions within the N gene of the RIT4385 vaccine.
- MeSH
- epitelové buňky MeSH
- farmaceutická technologie metody MeSH
- hydrocefalus patologie virologie MeSH
- krysa rodu rattus MeSH
- kultivace virů metody MeSH
- kultivované buňky MeSH
- kuřecí embryo MeSH
- modely nemocí na zvířatech MeSH
- novorozená zvířata MeSH
- plakové testy MeSH
- populační dynamika MeSH
- vakcína proti příušnicím aplikace a dávkování imunologie MeSH
- virová nálož MeSH
- virulence MeSH
- virus příušnic růst a vývoj imunologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
In virological practice, the pre-concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time-consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14 000 x g, and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real-time PCR. The pIs of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification.
- MeSH
- isoelektrická fokusace metody MeSH
- izoelektrický bod MeSH
- koncentrace vodíkových iontů MeSH
- koně MeSH
- kultivace virů metody MeSH
- Orthomyxoviridae chemie růst a vývoj izolace a purifikace MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody MeSH
- prasata MeSH
- sacharosa MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- MeSH
- elektronová mikroskopie metody normy využití MeSH
- imunologické testy metody normy využití MeSH
- klinické laboratorní techniky normy využití MeSH
- kultivace virů metody normy využití MeSH
- polymerázová řetězová reakce metody normy využití MeSH
- sérologické testy metody normy využití MeSH
- viry izolace a purifikace MeSH
- MeSH
- kultivace virů metody MeSH
- kultivační média chemie MeSH
- virus japonské encefalitidy růst a vývoj MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- infekce onkogenními viry diagnóza MeSH
- kultivace virů metody MeSH
- lidé MeSH
- onkogenní viry MeSH
- Check Tag
- lidé MeSH
