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  • 1
    Keywords: PEPTIDE ; RECEPTOR ; CELLS ; EXPRESSION ; CELL ; Germany ; PROTEIN ; PROTEINS ; MONOCLONAL-ANTIBODY ; LIGAND ; MOTIFS ; FAMILY ; BINDING ; SEQUENCE ; SEQUENCES ; ALPHA ; ACID ; ACIDS ; antibodies ; antibody ; MUTANT ; VECTOR ; VIRUS-LIKE PARTICLES ; EXTRACELLULAR-MATRIX ; SURFACE ; BETA ; CRYSTAL-STRUCTURE ; PEPTIDES ; LIGANDS ; MONOCLONAL-ANTIBODIES ; INTEGRIN ; EPITOPE ; AMINO-ACIDS ; STRUCTURAL-CHANGES ; RECEPTORS ; CELL-SURFACE RECEPTOR ; MATRIX ; SUBSET ; MOTIF ; VIRIONS ; INSECT CELLS ; ADENOVIRUS RECEPTOR ; ALPHA(V)BETA(3) INTEGRIN ; COXSACKIEVIRUS A9 ; MOUTH-DISEASE VIRUS ; MURINE POLYOMAVIRUS ; POLYOMAVIRUS ; PROTEIN VP1
    Abstract: Integrins are a family of cell surface proteins that function as receptors for extracellular matrix ligands and for some viruses. A subset of integrins recognises peptide sequences containing arginine-glycine-aspartic acid (RGD) motifs as ligands. The B-lymphotropic polyomavirus (LPV) has a non-enveloped capsid that recognises a sialylated cell surface receptor. To change the receptor binding specificity we have replaced sets of three amino acids in three predicted surface loops of the major capsid protein VP1 of the B-lymphotropic polyomavirus LPV by RGD. Ten mutants gave rise to the expected 40 kDa VP1 protein upon expression from a baculovirus vector in insect cells. Five of the VP1 mutants representing all three surface loops have retained the ability to spontaneously assemble to capsids in the nuclei of the insect cells. Structural changes of the mutant capsid surface were shown by differential reactivity with a set of 7 neutralising monoclonal antibodies that recognise conformational surface epitopes of wildtype LPV virions. In addition all mutant capsids had lost specific binding to the LPV receptor. Three mutant capsids of one loop (BC) showed specific binding to alphavbeta3 integrin but not to integrins alphavbeta5, alphavbeta6, or to alphaIIbbeta3 known also to recognise RGD containing peptide sequences. This selective binding of the mutant capsids could be inhibited by synthetic peptides that specifically bind to alphavbeta3 integrin with IC50 values between 10 and 40 nM
    Type of Publication: Journal article published
    PubMed ID: 15290355
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  • 2
    Keywords: RECEPTOR ; APOPTOSIS ; EXPRESSION ; GROWTH ; GROWTH-FACTOR ; Germany ; TYROSINE KINASE ; EXPOSURE ; PROTEIN ; ACTIVATION ; INFECTION ; INDUCTION ; KERATINOCYTES ; DYNAMICS ; MOLECULE ; PLASMA ; MEMBRANE ; STRESS ; human papillomavirus ; TYPE-16 ; EPITHELIAL-CELL LINE ; GOLGI-APPARATUS ; MHC CLASS-I ; PLASMA-MEMBRANE ; RECEPTORS ; HUMAN FORESKIN KERATINOCYTES ; keratinocyte ; phosphatidylcholine ; plasma membrane ; CERVICAL-CANCER WORLDWIDE ; CHOLESTEROL EFFLUX ; CTP-PHOSPHOCHOLINE CYTIDYLYLTRANSFERASE ; CYCLODEXTRIN ; SPHINGOMYELIN
    Abstract: The E5 protein of the human papillomavirus type 16 is a small protein found associated to membranes, mainly in the Golgi apparatus, and expressed in the early stages of viral infection. Its expression modifies the cell response towards growth factors and stress exposures, and also blocks the surface expression of MHC molecules. A global explanation for these multiple effects is hitherto not available. Here we present data showing that the expression of HPV16-E5 increases the amount of free cholesterol readily extractable from the plasma membrane, without altering the total cholesterol content. In addition, HPV16-E5 modifies the composition of the cell membranes, increasing the synthesis rate of phosphatidylcholine and phosphatidylserine, while diminishing that of phosphatidylglycerol. We propose that these changes in the lipid composition of the membrane are the central effect of HPV16-E5 on the cell. The multiple and apparently disconnected effects of HPV16-E5 on tyrosine-kinase receptors, induction of the apoptosis and impairment of MHC trafficking could follow the initial alteration on the membrane composition
    Type of Publication: Journal article published
    PubMed ID: 15503216
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  • 3
    Abstract: HPV types with high viral load are associated with cervical abnormalities. However, viral load measurements and concordance of HPV loads and viral mRNA have not been demonstrated for all high-risk/possibly high-risk (HR-/pHR-)HPV types in cervical cancer (CxCa). Especially, the biological role of co-infecting HR-/pHR-HPV types with low viral load has not been thoroughly investigated. Using BSGP5+/6+-PCR/MPG genotyping, we analyzed viral loads for all currently defined 51 mucosal HPV types in 74 cervical smears from patients with CxCa and compared this data with HPV DNA and mRNA status in these patients' corresponding CxCa tissues. All cervical smear/tissue pairs were HPV DNA+. Overall HPV type agreement within pairs was 99% (complete agreement in 50%, partial agreement in 49%, and complete disagreement in 1% of cases). The proportion of multiple HPV types was significantly higher in smears compared to tissues (p〈0.0001). High load HPV infections (〉1 copy/cell) were found in 88% of HPV DNA+ smears, and were significantly associated with the presence of respective HPV DNA (kappa=0.685, CI: 0.567-0.803), and HPV mRNA (kappa=0.693, CI: 0.566-0.820) in CxCa tissues. In total, 93% (67/72) of high load HR-/pHR-HPV infections identified in smears were also present in corresponding CxCa tissues, and 93% (62/67) of these were HPV mRNA+. On the other hand, 78% (42/54) of low load HR-/pHR-HPV infections identified in smears were not detectable in tissues, including 11 out of 15 low load HPV16 infections. This data demonstrates that the presence of high HPV loads in CxCa smears predicts biologically active HR-/pHR-HPV types in tumor tissues.
    Type of Publication: Journal article published
    PubMed ID: 28717857
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  • 4
    Keywords: RECEPTOR ; CELLS ; EXPRESSION ; GROWTH ; GROWTH-FACTOR ; CELL ; FACTOR RECEPTOR ; human ; TOXICITY ; GENE ; PROTEIN ; PROTEINS ; MECHANISM ; mechanisms ; BINDING ; BOVINE PAPILLOMAVIRUS ; papillomavirus ; ACID ; ACIDS ; MUTANT ; DIFFERENCE ; MEMBRANE ; human papillomavirus ; TYPE-16 ; WILD-TYPE ; LOCALIZATION ; NUCLEUS ; ADHESION ; CELL-ADHESION ; TRANSFORMATION ; EPITHELIAL-CELL LINE ; HUMAN KERATINOCYTES ; HUMAN-PAPILLOMAVIRUS ; EPITHELIAL-CELLS ; MORPHOLOGY ; ONCOPROTEIN ; SITE-DIRECTED MUTAGENESIS ; KINETICS ; AMINO-ACIDS ; RECEPTORS ; mutagenesis ; MEMBRANES ; Bcl-2 ; ACTIN CYTOSKELETON ; FINLAND ; CELLULAR-TRANSFORMATION ; E5 PROTEIN ; FACTOR RECEPTORS ; INTRACELLULAR MEMBRANES ; JUNCTIONAL INTERCELLULAR COMMUNICATION ; PORE-FORMING PROTEIN ; RECEPTOR ACTIVATION ; VACUOLAR H+-ATPASE
    Abstract: Human papillomavirus type 16 E5 protein contributes to cellular transformation by increasing the mitogenic stimulus from growth factor receptors to the nucleus. In order to study the biological mechanisms of the E5 protein we performed site-directed mutagenesis of the E5 gene. Wild-type as well as mutant E5 proteins were transiently expressed in human cervical epithelial cells, and cell morphology, expression of proteins involved in cell adhesion, and localization of the different proteins were studied. Little differences in cell morphology or expression kinetics were observed between the different E5 proteins, except for relocalization of a mutant E5 protein where a hydrophobic leucine membrane anchor was mutated to positively charged amino acids. This mutant E5 protein localized to lamellipodia, which are motility-associated structures at the leading edge of motile cells. In our experimental conditions, 100% of E5-expressing epithelial cells died by four days of expression, possibly due to toxicity or disturbance of the membrane compartment by the E5 protein. Most interestingly, a remarkable colocalization of the E5 protein with the Bcl-2 antiapoptotic protein on intracellular membranes was established
    Type of Publication: Journal article published
    PubMed ID: 15593417
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  • 5
    Keywords: CELLS ; CELL ; INHIBITION ; DRUG ; TIME ; INFECTION ; CONTRAST ; CYCLE ; ENTRY ; PARTICLES ; virus ; MEMBRANE ; BETA ; cholesterol ; lipids ; LIPID RAFTS ; ENVELOPED VIRUSES ; HUMAN-IMMUNODEFICIENCY-VIRUS ; LIFE ; MICRODOMAINS ; membrane rafts ; MEDIATED ENDOCYTOSIS ; CD ; SERINE PALMITOYLTRANSFERASE
    Abstract: Lipid rafts are involved in the life cycle of many viruses. In this study, we investigated the role of lipids in the life cycle of vesicular stomatitis virus (VSV). Cholesterol depletion by pretreatment of BHK cells or VSV particles with methyl-beta-cyclodextrin (M beta CD), a cholesterol-sequestering drug, inhibited the production of VSV dramatically. This effect was reversible, and virus production was restored by the addition of cholesterol, indicating that the reduction was caused by the loss of cholesterol in the cell membrane and virus, respectively. Cholesterol depletion at the adsorption stage also reduced the production of VSV significantly, but in contrast, only had a limited effect on virus production at the post-entry stage. Inhibition of sphingomyelin by myriocin treatment only showed a minor effect on VSV production. However, reduction of cholesterol and sphingomyelin at the same time dramatically reduced VSV production, showed a significant synergistic effect. These results suggest that lipid rafts play an important role in the life cycle of VSV
    Type of Publication: Journal article published
    PubMed ID: 19288237
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  • 6
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The susceptibility of congenitally anemic W/Wv mice to infection with murine cytomegalovirus (MCMV) was examined. W/Wv mice showed a higher mortality rate and shorter survival time after MCMV infection than did their +/+ littermate mice. In addition, W/Wv mice showed a lower plaque-forming unit (PFU) per 50% lethal dose (LD50) and produced higher titers of infectious virus in various organs. The mortality rate and survival time of W/Wv mice which received a bone marrow graft 4 weeks before infection was completely restored to the level for +/+ mice, suggesting the importance of the cells of myeloid origin. Although natural killer (NK) activity of W/Wv mice was comparable to that of +/+ mice before infection, marked reduction was observed after MCMV infection. Furthermore, OK-432 treatment failed to enhance NK activity of W/Wv mice. Impaired NK response was also completely restored by bone marrow grafting 4 weeks before infection. The level of serum interferon (IFN) of infected or uninfected W/Wv mice was comparable to that of +/+ mice. Therefore, impaired NK inducibility seems to be responsible, at least in part, for the high susceptibility of W/Wv mice to MCMV infection.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Archives of virology 101 (1988), S. 255-259 
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Incoulation of lambs with a New Zealand strain of orf virus (NZ 2) failed to protect them against subsequent infection with the Lister strain of vaccinia virus. Similarly, in the reciprocal test, vaccinia virus failed to protect against subsequent orf virus infection. Inoculation with either orf virus or vaccinia virus alone afforded protection against reinfection with the same virus. These results have relevance to the use of vaccinia virus gene vectors in sheep.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Archives of virology 101 (1988), S. 261-261 
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A 6008 base pair fragment of the vaccinia virus DNA containing the gene for the precursor of the major core protein 4 a, which has been designated P4 a, was sequenced. A long open reading frame (ORF) encoding a protein of molecular weight 102,157 started close to the position where the P4 a mRNA had been mapped. Analysis of the mRNA by S1 nuclease mapping and primer extension indicated that the 5′ end defined by the former method is not the true 5′ end. This suggests that the P4 a coding region is preceded by leader sequences that are not derived from the immediate vicinity of the gene, similar to what has been reported for another late vaccinia virus mRNA. The sequenced DNA contained several further ORFs on the same, or opposite DNA strand, providing further evidence for the close spacing of protein-coding sequences in the viral genome.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Eleven European isolates of Suid herpesvirus type 1 (SHV-1) were compared by restriction fragment pattern analyses and Southern blot hybridization using different genomic probes. The presence of strain discriminative 4 major genome types and several subtypes as well as the molecular distinctions between these were demonstrated. Evolutionary relationships between the different types are suggested on the basis of homologous restriction fragment length polymorphisms.
    Type of Medium: Electronic Resource
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