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  • 1
    Keywords: CELLS ; EXPRESSION ; carcinoma ; CELL ; Germany ; human ; GENE ; GENE-EXPRESSION ; GENES ; PROTEIN ; PROTEINS ; MOLECULES ; COMPLEX ; COMPLEXES ; KERATINOCYTES ; BOVINE PAPILLOMAVIRUS ; DOWN-REGULATION ; GROWTH-FACTOR RECEPTOR ; papillomavirus ; MOLECULE ; MHC ; TRANSPORT ; IDENTIFICATION ; LESIONS ; gene expression ; DESIGN ; DIFFERENCE ; PLASMA ; MEMBRANE ; STRESS ; SPECTROMETRY ; human papillomavirus ; TYPE-16 ; MASS-SPECTROMETRY ; SURFACE ; CLASS-I ; EPITHELIAL-CELL LINE ; GOLGI-APPARATUS ; HUMAN-PAPILLOMAVIRUS ; MHC class I ; MHC CLASS-I ; CARCINOMAS ; DIFFERENTIAL EXPRESSION ; MEMBRANE PROTEIN ; HaCaT ; MEMBRANE-PROTEIN ; GEL-ELECTROPHORESIS ; MASSES ; E5 PROTEIN ; CERVICAL NEOPLASIA
    Abstract: Membrane proteins differentially expressed in human papillomavirus type 16 (HPV-16) E5-transfected HaCaT cells have been identified. Membrane proteins were isolated and separated by two-dimensional gel electrophoresis. Spots showing quantitative differences between E5-transfected and control cells were extracted and the proteins were identified by nanoelectrospray ionization mass spectrometry. A total of 24 spots was analysed. Among the proteins showing differential expression, a decreased amount of calnexin and increased expression of hsp70, proteins both involved in maturation and transport of MHC class I complexes to the plasma membrane, were noticed. These findings correlate with the decreased surface expression of MHC class I molecules described in E5-expressing cells, HPV-positive cervical lesions and cervical carcinomas. These results stress the value of the proteomic approach, as used here in the experimental design, which allows the correlation of changes in host gene expression with biological functions of viral genes
    Type of Publication: Journal article published
    PubMed ID: 15166425
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  • 2
    Keywords: CELLS ; CELL ; Germany ; human ; SITE ; SITES ; DISTINCT ; GENE ; GENES ; GENOME ; transcription ; cell line ; FAMILY ; SEQUENCE ; SEQUENCES ; TARGET ; virus ; VECTOR ; PROMOTER ; NUMBER ; CELL-LINE ; leukemia ; LINE ; PCR ; HUMAN GENOME ; REGION ; REGIONS ; ONCOGENE ; SELECTION ; foamy virus ; GENE-THERAPY ; HEMATOPOIETIC STEM-CELLS ; RETROVIRAL VECTORS ; DNA INTEGRATION ; HUMAN-IMMUNODEFICIENCY-VIRUS ; CPG-ISLANDS ; INTEGRATION ; FAMILIES ; CPG ISLANDS ; HIV ; SEVERE COMBINED IMMUNODEFICIENCY ; HIGH-THROUGHPUT ; PROMOTER REGION ; IMMUNODEFICIENCY VIRUS ; LEUKEMIA-VIRUS ; HUMAN-GENOME ; TRANSCRIPTION START REGIONS ; HIV INTEGRASE ; RETROTRANSPOSITION ; RETROVIRAL INTEGRATION ; SITE SELECTION
    Abstract: Integration-site selection by retroviruses and retroviral vectors has gained increased scientific interest. Foamy viruses (FVs) constitute a unique subfamily (Spumavirinae) of the family Retroviridae, for which the integration pattern into the human genome has not yet been determined. To accomplish this, 293 cells were transduced with FV vectors and the integration sites into the cellular genome were determined by a high-throughput method based on inverse PCR. For comparison, a limited number of murine leukemia virus (MLV) and human immunodeficiency virus (HIV) integration sites were analysed in parallel. Altogether, 628 FV, 87 HIV and 141 MLV distinct integration sites were mapped to the human genome. The sequences were analysed for RefSeq genes, promoter regions, CpG islands and insertions into cellular oncogenes. Compared with the integration-site preferences of HIV, which strongly favours active genes, and MLV, which favours integration near transcription-start regions, our results indicate that FV integration has neither of these preferences. However, once integration has occurred into a transcribed region of the genome, FVs tend to target promoter-close regions, albeit with less preference than MLV. Furthermore, our study revealed a palindromic consensus sequence for integration, which was centred on the virus-specific, four-base-duplicated target site. In summary, it is shown that the integration pattern of FVs appears to be unique compared with those of other retroviral genera
    Type of Publication: Journal article published
    PubMed ID: 16603537
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  • 3
    Keywords: CANCER ; CELLS ; BLOOD ; CELL ; Germany ; human ; IN-VIVO ; MODEL ; VIVO ; SYSTEM ; incidence ; RISK ; HYBRIDIZATION ; TUMORS ; DNA ; INFECTION ; IMPACT ; CARCINOGENESIS ; RAT ; animals ; tumour ; SKIN ; papillomavirus ; TARGET ; TRANSGENIC MICE ; IN-SITU ; skin cancer ; LINE ; E6 ; BENIGN ; STEM-CELLS ; TARGET-CELLS ; INVOLVEMENT ; SQUAMOUS-CELL CARCINOMAS ; TARGETS ; INFECTIONS ; SKIN-CANCER ; FOLLICLE ; in situ hybridization ; HUMAN CANCER ; HAIR FOLLICLE ; RECIPIENTS ; LIFE-CYCLE ; development ; chemical carcinogenesis ; PERSISTENCE ; HUMAN CANCERS ; CANCERS ; in vivo ; PREDICTOR ; animal ; microbiology ; viral ; virology ; Mothers ; biotechnology ; GENOMES
    Abstract: The high incidence of multiple wart formation and skin cancer in organ-transplant recipients, as well as the question of an involvement of papillomaviruses in a variety of human cancers, require a model system for papillomavirus infections in immunocompetent animals. Such an in vivo model is represented by the multimammate rat Mastomys coucha, which is infected with Mastomys natalensis papillomavirus (MnPV). MnPV primarily induces benign skin tumours, such as papillomas and keratoacanthomas. Here, the incidence of MnPV infections in different skin areas and various organs is described. In situ hybridization showed that hair follicle cells were positive for viral DNA and that the amount of MnPV in normal skin may be considered a predictor for the development of skin tumours. MnPV infection is not restricted to the skin, but can also be detected in inner organs. As the blood and the lymphatic system were temporarily also found to be virus-positive, a haematogenic propagation of MnPV can be assumed. However, MnPV is apparently not transmitted through the germ line, as fetuses and newborns lack viral DNA, despite infection of their mothers. In conclusion, M. coucha is not only useful to study papillomavirus-induced skin carcinogenesis, but may also serve as a model to identify additional, still unknown target cells of papillomavirus infections and the potential pathological impact
    Type of Publication: Journal article published
    PubMed ID: 17872518
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  • 4
    Keywords: CELLS ; CELL ; human ; MODEL ; GENOME ; PROTEIN ; PROTEINS ; SAMPLE ; SAMPLES ; TUMORS ; PATIENT ; DNA ; INFECTION ; CARCINOGENESIS ; tumour ; SKIN ; BINDING ; E7 ; OPEN READING FRAME ; SEQUENCES ; skin carcinogenesis ; PCR ; SWEDEN ; HPV ; INFECTIONS ; NONMELANOMA SKIN CANCERS ; real-time PCR ; BIOPSY ; RETINOBLASTOMA PROTEIN ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; REAL-TIME ; E6 PROTEINS ; microbiology ; ENGLAND ; HUMAN PAPILLOMAVIRUSES ; retinoblastoma ; BIOPSIES ; E7 PROTEIN ; viral ; virology ; biotechnology ; GENOMES ; HPV types ; HPV DNA ; HEALTHY SKIN ; MALIGNANT-TRANSFORMATION
    Abstract: Two novel human papillomaviruses (HPVs), HPV93 and HPV96, with genomes of 7450 and 7438 bp, respectively, are described. The Ll open reading frame of HPV93 showed highest identity to HPV24 (79%) and that of HPV96 had highest identity to HPV92 (71 %). Real-time PCR for HPV92, 93 and 96 on stripped biopsies from tumours and healthy skin from 269 immunocompetent patients found HPV DNA in 2.6 % of tumours and in 0.4 % of healthy skin samples. Double infections were observed in two tumours. HPV92 was detected in four, HPV93 in two and HPV96 in three tumours. The range of viral loads spanned from one copy per 45 cells to one copy per 10 000 cells. The E7 proteins of HPV92, 93 and 96 were found to bind the retinoblastoma protein (pRb). These results suggest a possible role for these HPV types in skin carcinogenesis that deserves further study,
    Type of Publication: Journal article published
    PubMed ID: 17412976
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  • 5
    Keywords: carcinoma ; CELL ; EXPOSURE ; POPULATION ; HYBRIDIZATION ; PATIENT ; INFECTION ; SKIN ; FREQUENCIES ; LESIONS ; ASSAY ; AGE ; HPV ; DIVERSITY ; HUMAN-PAPILLOMAVIRUS DNA ; SQUAMOUS-CELL CARCINOMA ; NETHERLANDS ; squamous cell carcinoma ; INDIVIDUALS ; PREVALENCE ; RENAL-TRANSPLANT RECIPIENTS ; immunosuppression ; CELL CARCINOMA ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; hair ; NONMELANOMA SKIN-CANCER ; SQUAMOUS-CELL ; SUN EXPOSURE ; virology ; biotechnology ; GENERAL-POPULATION ; HEALTHY SKIN ; HAIRS ; KERATOSES ; SEROREACTIVITY
    Abstract: Betapapillomavirus (betaPV) infections are often associated with squamous-cell carcinoma (SCC) and the prevalence of betaPV infections in (immunosuppressed) SCC patients is known to be high. The distribution and possible associated factors of betaPV infections in the general population, however, are largely unknown. To address this issue, betaPV infection was studied in 1405 SCC-free immunocompetent (n=845) and immunosuppressed (n=560) individuals from six countries of different latitudes. A standard study protocol was used to obtain information about age, sex, UV-irradiation and skin type, and from all participants eyebrow hairs were collected for detection and genotyping of 25 established betaPV types using the PM-PCR reverse hybridization assay (RHA) method. The frequency of betaPV-positive participants ranged from 84 to 91% in the immunocompetent population with HPV23 as the most prevalent type, and from 81 to 98% in the immunosuppressed population with HPV23 as the most or the second most prevalent type. The median number of infecting betaPV types ranged from four to six in the immunocompetent and from three to six in the immunosuppressed population. Increasing age in the immunocompetent participants and (duration of) immunosuppression in the immunosuppressed patients were associated with betaPV infection. In both groups, sex, skin phototype, sunburns and sun-exposure were not consistently associated with betaPV infection. This study demonstrates that betaPV infections are also highly prevalent in SCC-free individuals, with similar HPV types prevailing in both immunocompetent and immunosuppressed persons. Age and (duration of) immunosuppression were identified as betaPV infection-associated factors, whereas characteristics related to sun exposure and skin type were not
    Type of Publication: Journal article published
    PubMed ID: 19321753
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  • 6
    Keywords: Germany ; human ; CLASSIFICATION ; TISSUE ; DNA ; papillomavirus ; LESIONS ; human papillomavirus ; HPV ; DIVERSITY ; HUMAN-PAPILLOMAVIRUS ; SQUAMOUS-CELL CARCINOMA ; PREVALENCE ; NONMELANOMA SKIN CANCERS ; RENAL-TRANSPLANT RECIPIENTS ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; NUCLEOTIDE-SEQUENCE ; IMMUNOCOMPETENT INDIVIDUALS ; virology ; biotechnology ; KERATOSIS ; PAPILLOMAVIRUS TYPES
    Abstract: Seven novel human papillomavirus (HPV) types were isolated and characterized. HPV 94 is related most closely to HPV 10 and belongs to the genus Alphapapillomavirus, whereas HPV 98, HPV 99, HPV 100, HPV 104, HPV 105 and HPV 113 all belong to the genus Betapapillomavirus. These HPV types were isolated from and demonstrated in cutaneous tissue, but HPV 98, HPV 100, HPV 104 and HPV 113 were also detected in malignant oesophageal and oral lesions. The general prevalence of these HPV types in lesions is infrequent
    Type of Publication: Journal article published
    PubMed ID: 19386784
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  • 7
    Keywords: CELLS ; CELL ; COMBINATION ; Germany ; GENE ; GENES ; GENOME ; PROTEINS ; COMPLEX ; COMPLEXES ; SKIN ; papillomavirus ; ACID ; EVOLUTION ; L1 ; PHYLOGENETIC ANALYSIS ; AMINO-ACID ; hair ; LONG ; adaptation ; GENOMES ; SEQUENCE DATA ; EEPV
    Abstract: Knowledge about biological diversity is the prerequisite to reliably reconstruct the evolution of pathogens such as papillomaviruses (PV). However, complete genomes of non-human PV have only been cloned and sequenced from 8 out of 18 orders within the Placentalia, although the host-specific variety of PV is considered much larger. We isolated and sequenced the complete genome of the first insectivoran PV type from hair follicle cells of the European hedgehog (Erinaceus europaeus), designated EHPV. We conducted phylogenetic analyses (maximum-likelihood criterion and Bayesian inference) with the genomic information of a systematically representative set of 67 PV types including EHPV As inferred from amino acid sequence data of the separate genes E1, E2 and L1 as well as of the gene combination E6-E7-E1-E2-L1, EHPV clustered within the beta-gamma-pi-zeta-PV supertaxon and constituted the closest relative of genus Betapapillomavirus infecting primates. Beside the typical organization of the PV genome, EHPV exhibited a 1172 lop, non-coding region between the E2 and the L2 open reading frames. This trait has been previously described for the only distantly related Lambdapapillomavirus, but a common evolutionary origin of both non-coding regions is unlikely. Our results underscore the modular organization of the PV genome and the complex natural history of PV
    Type of Publication: Journal article published
    PubMed ID: 19218207
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  • 8
    Keywords: SPECTRA ; CANCER ; carcinoma ; CELL ; Germany ; human ; EXPOSURE ; HISTORY ; POPULATION ; RISK ; GENOME ; radiation ; RESPONSES ; DNA ; INFECTION ; CARCINOGENESIS ; SKIN ; papillomavirus ; antibodies ; antibody ; LESIONS ; WOMEN ; MEN ; RISK FACTOR ; human papillomavirus ; HPV ; HUMAN-PAPILLOMAVIRUS ; SQUAMOUS-CELL CARCINOMA ; NETHERLANDS ; squamous cell carcinoma ; INDIVIDUALS ; sensitivity ; NATURAL-HISTORY ; RENAL-TRANSPLANT RECIPIENTS ; glutathione-S-transferase ; SERUM ; CELL CARCINOMA ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; development ; RISK-FACTOR ; SQUAMOUS-CELL ; SUN EXPOSURE ; virology ; SEROPREVALENCE ; biotechnology ; CUTANEOUS HUMAN PAPILLOMAVIRUSES ; CONFIDENCE ; SCC ; PAPILLOMAVIRUS TYPES
    Abstract: Solar UV radiation is the main risk factor for cutaneous squamous cell carcinoma (SCC), but infections with skin human papillomavirus (HPV) types have also been linked to the development of SCC. Little is known about the natural history of these infections and whether the seroprevalence of skin HPV types is affected by ambient or individual levels of sun exposure. This study investigated this by analysing sera for antibodies to 26 skin HPV types from five phylogenetic genera obtained from 807 healthy individuals from the Netherlands, Italy and Australia, countries with strong differences in sunlight intensity. Overall HPV seroprevalence, was similar across the three countries (50-57% for beta-HPV types, 40-48% for gamma-HPV types), and the most frequent beta-HPV and gamma-HPV types were the same in all countries. The highest seroprevalences; for 24 of the 26 skin HPV types were observed in Italy (114 types) and Australia (ten types). Seroprevalence among men was generally higher than among women, and the male sex was significantly associated with both beta-HPV [odds ratio (OR) 2.81, 95 % confidence interval (CI) 1.64-4.821 and gamma-HPV (OR 2.42, 95% CI 1.40-4.18) antibodies in Australia. The only measure of sun sensitivity or UV exposure significantly associated with skin HPV seroprevalence was found for weekend sun exposure in Australia and beta-HPV antibodies. It was concluded that type spectra and HPV seroprevalence are similar in countries with different sunlight intensity, and that levels of UV exposure do not play a strong role in the development of skin HPV antibodies in this study population
    Type of Publication: Journal article published
    PubMed ID: 19386782
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  • 9
    Abstract: Human papillomaviruses (HPVs) of the genus Betapapillomavirus appear to be involved in the early stages of skin cancer development, since both the prevalence and viral load are higher in precancerous actinic keratoses than in skin cancers. Interleukin-8 (IL-8) is an inflammatory cytokine that serves to alert the surrounding tissue after UV-induced damage. We examined the effects of the E2, E6 and E7 proteins of HPV8 and the E6 proteins of various HPV genotypes on IL-8 secretion from primary keratinocytes. HPV5 and HPV8 E6 showed the highest downregulation of basal IL-8 secretion. HPV8 E6 also negatively modulated IL-8 mRNA expression and protein secretion upon UVB irradiation. The downregulation of IL-8 in actinic keratoses may weaken the response to UV-induced damage and thus favour the accumulation of UVB-induced mutations.
    Type of Publication: Journal article published
    PubMed ID: 20007354
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  • 10
    Keywords: ACID, AMINO-ACID, animals, CANCER, CELL, CELLS, cluster analysis, DNA,Viral/chemistry/genetics, Evol
    Abstract: A series of papillomavirus (PV) types have been isolated from different rodent species, and most of them belong to the genus Pipapillomavirus. We isolated and sequenced the complete genome of a novel PV type (designated RnPV) from the oral cavity of the Norway rat (Rattus norvegicus), as well as an L1 gene fragment from hair-follicle cells of the European beaver (Castor fiber). As inferred from amino acid sequence data, RnPV clustered within the beta+gamma+pi+Xi-PV supertaxon as a member of the genus Pipapillomavirus. The closest relatives of RnPV were McPV-2 and MmPV, and time estimates indicated that the genus Pipapillomavirus originated in the late Cenozoic era. The close relationship of RnPV to other murid PV types supports the hypothesis of co-divergence between members of the genus Pipapillomavirus and their hosts. However, the derived Neogene origin of the genus Pipapillomavirus is much younger than has been considered for the Rodentia as the primary hosts, indicating that alternative interpretations of the phylogenetic trees should be conceived.
    Type of Publication: Journal article published
    PubMed ID: 19605590
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