Tick-borne encephalitis virus (TBEV) is a causative agent of tick-borne encephalitis (TBE), one of the most important human infections involving the central nervous system. Although effective vaccines are available on the market, they are recommended only in endemic areas. Despite many attempts, there are still no specific antiviral therapies for TBEV treatment. Previously, we synthesized a series of uridine derivatives of 2-deoxy sugars and proved that some compounds show antiviral activity against viruses from the Flaviviridae and Orthomyxoviridae families targeting the late steps of the N-glycosylation process, affecting the maturation of viral proteins. In this study, we evaluated a series of uridine derivatives of 2-deoxy sugars for their antiviral properties against two strains of the tick-borne encephalitis virus; the highly virulent TBEV strain Hypr and the less virulent strain Neudoerfl. Four compounds (2, 4, 10, and 11) showed significant anti-TBEV activity with IC50 values ranging from 1.4 to 10.2 μM and low cytotoxicity. The obtained results indicate that glycosylation inhibitors, which may interact with glycosylated membrane TBEV E and prM proteins, might be promising candidates for future antiviral therapies against TBEV.
- MeSH
- antivirové látky chemie farmakologie MeSH
- deoxycukry chemie farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- molekulární struktura MeSH
- nádorové buněčné linie MeSH
- plakové testy MeSH
- proteosyntéza účinky léků MeSH
- uridin analogy a deriváty chemie farmakologie MeSH
- viry klíšťové encefalitidy účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
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
BACKGROUND: The clinical course of tick-borne encephalitis (TBE), a disease caused by TBE virus, ranges from asymptomatic or mild influenza-like infection to severe debilitating encephalitis or encephalomyelitis. Despite the medical importance of this disease, some crucial steps in the development of encephalitis remain poorly understood. In particular, the basis of the disease severity is largely unknown. METHODS: TBE virus growth, neutralizing antibody response, key cytokine and chemokine mRNA production and changes in mRNA levels of cell surface markers of immunocompetent cells in brain were measured in mice with different susceptibilities to TBE virus infection. RESULTS: An animal model of TBE based on BALB/c-c-STS/A (CcS/Dem) recombinant congenic mouse strains showing different severities of the infection in relation to the host genetic background was developed. After subcutaneous inoculation of TBE virus, BALB/c mice showed medium susceptibility to the infection, STS mice were resistant, and CcS-11 mice were highly susceptible. The resistant STS mice showed lower and delayed viremia, lower virus production in the brain and low cytokine/chemokine mRNA production, but had a strong neutralizing antibody response. The most sensitive strain (CcS-11) failed in production of neutralizing antibodies, but exhibited strong cytokine/chemokine mRNA production in the brain. After intracerebral inoculation, all mouse strains were sensitive to the infection and had similar virus production in the brain, but STS mice survived significantly longer than CcS-11 mice. These two strains also differed in the expression of key cytokines/chemokines, particularly interferon gamma-induced protein 10 (IP-10/CXCL10) and monocyte chemotactic protein-1 (MCP-1/CCL2) in the brain. CONCLUSIONS: Our data indicate that the genetic control is an important factor influencing the clinical course of TBE. High neutralizing antibody response might be crucial for preventing host fatality, but high expression of various cytokines/chemokines during TBE can mediate immunopathology and be associated with more severe course of the infection and increased fatality.
- MeSH
- buněčná imunita imunologie MeSH
- centrální nervový systém patologie MeSH
- chemokiny biosyntéza MeSH
- cytokiny biosyntéza MeSH
- genotyp MeSH
- klíšťová encefalitida imunologie patologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- membránové proteiny biosyntéza MeSH
- messenger RNA biosyntéza genetika MeSH
- mozek - chemie fyziologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- náchylnost k nemoci MeSH
- neutralizující protilátky biosyntéza MeSH
- odolnost vůči nemocem MeSH
- plakové testy MeSH
- protilátky virové biosyntéza genetika MeSH
- virová nálož MeSH
- viry klíšťové encefalitidy * MeSH
- zánět patologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- dospělí MeSH
- Flavivirus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- neutralizační testy MeSH
- plakové testy MeSH
- protilátky virové krev MeSH
- virus západního Nilu imunologie izolace a purifikace růst a vývoj MeSH
- západonilská horečka epidemiologie krev MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Tahyna virus (TAHV), a mosquito-borne bunyavirus (California group), is frequently associated with inapparent or influenza-like (Valtice fever) infections in humans, rarely leading to atypical pneumonia or meningitis. Field TAHV strains exhibit a high variability in their biological properties with respect to virulence for laboratory mouse, temperature-sensitivity or character of plaques in cell culture. In consideration of the variations in the antigenic properties TAHV and its potential genetic variability, we analyzed complete nucleotide sequences of the small (S) and medium (M) genomic segments of field TAHV strains with different combinations of phenotypic markers. S segment was highly conservative in all analyzed TAHV strains. Within the M segment, the highest variability was observed in the G(C) gene encoding viral envelope protein and to a less extent also in the NSm gene. However, 5' and 3' non-coding regions of M segment, as well as in G(N) gene exhibited highly conservative pattern, indicating its functional importance, but minor or no role in the determination of biological properties of TAHV field strains.
- MeSH
- 3' nepřekládaná oblast MeSH
- 5' nepřekládaná oblast MeSH
- fylogeneze MeSH
- genetická variace MeSH
- genom virový MeSH
- kalifornská encefalitida virologie MeSH
- konzervovaná sekvence MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši MeSH
- plakové testy MeSH
- proteiny virového obalu genetika MeSH
- RNA virová genetika MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie MeSH
- shluková analýza MeSH
- virulence MeSH
- viry kalifornské encefalitidy genetika izolace a purifikace patogenita MeSH
- vysoká teplota MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Smallpox was declared eradicated in 1980. However recently, the need of agents effective against poxvirus infection has emerged again. In this paper, we report an original finding that two redox-modulating agents, the ethacrynic and alpha-lipoic acids (EA, LA), inhibit growth of vaccinia virus (VACV) in vitro. The effect of EA and LA was compared with those of beta-mercaptoethanol, DTT and ascorbic acid, but these agents increased VACV growth in HeLa G cells. The inhibitory effects of EA and LA on the growth of VACV were further confirmed in several cell lines of different embryonic origin, in epithelial cells, fibroblasts, macrophages and T-lymphocytes. Finally, we have analyzed the mechanism of action of the two agents. They both decreased expression of VACV late genes, as demonstrated by western blot analysis and activity of luciferase expressed under control of different VACV promoters. In contrast, they did not inhibit virus entry into the cell, expression of VACV early genes or VACV DNA synthesis. The results suggest new directions in development of drugs effective against poxvirus infection.
- MeSH
- antivirové látky farmakologie MeSH
- exprese genu účinky léků MeSH
- financování organizované MeSH
- HeLa buňky MeSH
- kultivované buňky MeSH
- kyselina ethakrynová farmakologie MeSH
- kyselina lipoová farmakologie MeSH
- lidé MeSH
- luciferasy metabolismus MeSH
- plakové testy MeSH
- replikace viru účinky léků MeSH
- reportérové geny MeSH
- virus vakcinie růst a vývoj účinky léků MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
Various processes of bacteriophage lambda development in Escherichia coli cells bearing either the whole lambda exo-xis region (with truncated, thus nonfunctional, exo and xis genes) or particular genes from this region were investigated. The presence of either the exo-xis region or the ea8.5 gene on a plasmid resulted in formation of fuzzy plaques by infecting phage. Both efficiency of plating and efficiency of lysogenization were decreased in such hosts. On the other hand, neither the efficiency of adsorption nor intracellular lytic development of the infecting phage (measured in one-step-growth experiments) was affected while significantly more host cells survived the infection, when containing the exo-xis region. Although no effects of the exo-xis region on the activity of the p (L) promoter was detected, this region contributed to a decreased transcription from the cII-stimulated promoters p (I), p (aQ) and p (E). These results, together with the results of measurement of efficiency of plating of phages bearing mutations in cI, cII and cIII genes on hosts containing the exo-xis region, strongly suggest that genes from this region (especially ea8.5) are involved in the regulation of bacteriophage lambda development at the stage of the lysis-vs.-lysogenization decision.
- MeSH
- bakteriofág lambda fyziologie genetika růst a vývoj MeSH
- bakteriolýza MeSH
- DNA-nukleotidyltransferasy genetika chemie MeSH
- Escherichia coli virologie MeSH
- exodeoxyribonukleasy genetika chemie MeSH
- lyzogenie MeSH
- mutace MeSH
- plakové testy MeSH
- regulace exprese virových genů MeSH
- virové proteiny genetika chemie metabolismus MeSH
A phage antibody display library of single chain fragment variables (scFv) was applied to develop anti-equid herpesvirus-1 (EHV-1) glycoprotein D (gD) neutralizing antibodies. To enrich for specific scFvs, the phage antibody library was panned against epitope derived from the N-terminal part of EHV-1 gD. Unique clones were differentiated by BstNI fingerprinting and further characterized by sequencing and immunoreactivity. The neutralizing effect of each clone was assessed by plaque reduction assay. Three clones with neutralizing effect were isolated.
- MeSH
- buněčné linie MeSH
- financování organizované MeSH
- fluorescenční mikroskopie MeSH
- herpesvirus 1 koní imunologie MeSH
- herpetické infekce imunologie veterinární MeSH
- imunoblotting MeSH
- imunoglobuliny - fragmenty imunologie MeSH
- kinetika MeSH
- koně MeSH
- molekulární sekvence - údaje MeSH
- nemoci koní virologie MeSH
- peptidová knihovna MeSH
- plakové testy MeSH
- proteiny virového obalu imunologie MeSH
- rekombinantní proteiny imunologie MeSH
- sekvence aminokyselin MeSH
- variabilní oblast imunoglobulinu imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
