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  • 1
    Keywords: CANCER ; tumor ; carcinoma ; human ; CLASSIFICATION ; EXPOSURE ; RISK ; SITE ; PROTEIN ; PROTEINS ; TUMORS ; PATIENT ; DNA ; INFECTION ; FAMILY ; RISK-FACTORS ; SKIN ; MR ; papillomavirus ; ASSOCIATION ; antibodies ; IN-SITU ; risk factors ; PATHOGENESIS ; human papillomavirus ; VIRUS-LIKE PARTICLES ; HUMAN-PAPILLOMAVIRUS ; CARCINOMAS ; case-control studies ; squamous cell carcinoma ; INDIVIDUALS ; sensitivity ; RENAL-TRANSPLANT RECIPIENTS ; basal cell carcinoma ; glutathione-S-transferase ; CELL CARCINOMA ; case-control study ; population-based case-control study ; ASSOCIATIONS ; case control studies ; INTERVAL ; TECHNOLOGY ; RISK-FACTOR ; CANCERS ; population-based ; IMMUNOCOMPETENT INDIVIDUALS ; E6 PROTEIN ; multiplex serology ; PLUCKED EYEBROW HAIRS
    Abstract: Background. Although infection with human papillomaviruses (HPVs) is a major risk factor for several epithelial cancers, an etiologic relationship between HPV and keratinocyte cancers, such as squamous cell carcinomas (SCCs) and basal cell carcinomas (BCCs), remains unclear. Methods: In a population-based case-control study of 252 SCC case patients, 525 BCC case patients, and 461 control subjects, we used multiplex serology to detect antibodies in plasma samples against 16 HPV types from phylogenetic genera alpha, beta, and mu. Multiplex serology is a new method that is based on fluorescent bead technology and allows simultaneous detection of antibodies against up to 100 different in situ affinity-purified recombinant HPV proteins. Data on sun sensitivity, outdoor exposure, and other risk factors for keratinocyte cancers were collected through personal interviews. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated via unconditional logistic regression models. Results: Overall, we detected HPV antibodies more frequently in SCC patients than in control subjects (OR = 1.6, 95% CI = 1.2 to 2.3), but we found no difference in HPV seropositivity between BCC case patients and control subjects (OR = 0.8, 95% CI = 0.6 to 1.1). Among HPV types, seropositivity to HPV types in genus beta (OR = 1.5,95% CI = 1.0 to 2.1), particularly HPV 5 (OR = 1.8,95% CI = 1.0 to 3.1), was associated with SCC risk. Individuals with tumors on chronically sun exposed sites were more likely to be seropositive for beta HPV types than individuals with SCC at other anatomic sites. The highest SCC risk was associated with positivity for multiple HPV types and, among individuals seropositive for HPV beta, a tendency to sunburn; however, the associations had limited statistical precision. Conclusions: Our findings support a role for HPV types from the genus beta in the pathogenesis of SCC
    Type of Publication: Journal article published
    PubMed ID: 16537831
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  • 2
    Keywords: Germany ; human ; screening ; PROTEINS ; RISK-FACTORS ; ANTIGEN ; BINDING ; antibodies ; virus ; ASSAY ; CERVICAL-CANCER ; human papillomavirus ; HUMAN-PAPILLOMAVIRUS ; GLUTATHIONE S-TRANSFERASE ; SERUM ; ELISA ; RE ; ASSAYS ; TECHNOLOGY ; MICROSPHERE IMMUNOASSAY ; multiplex serology ; Luminex ; polyvinylalcohol ; polyvinylpyrrolidone ; xMAP
    Abstract: Bead-based suspension array technology (xMAP, Luminex Corp.) permits the simultaneous analysis of antibodies with specificities for up to 100 different antigens in a single reaction and the high through-put screening Of LIP to 1000 sera per day. Therefore, this technology is becoming more and more popular for serological analyses. replacing ELISA techniques at least for epidemiological purposes. However, a major intrinsic problem of Luminex technology is that human sera may contain antibodies that directly bind to the beads, resulting in intolerably high non-specific background, The proportion of such "bead binders" in different serum panels frequently exceeds 5% and is therefore a severe problem. We screened for background inhibitors and found that serum pre-incubation with polyvinylalcohol, polyvinylpyrrolidone and a proprietary reagent (Super ChemiBlock, Chemicon) significantly reduced non-specific background, whereas use of Luminex SeroMap beads only partially solved the problem. (c) 2005 Elsevier B.V. All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 16406059
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  • 3
    Keywords: OPTIMIZATION ; PEPTIDE ; CANCER ; INHIBITOR ; evaluation ; Germany ; human ; THERAPY ; screening ; PROTEIN ; PROTEINS ; DRUG ; INFECTION ; BINDING ; papillomavirus ; antibodies ; antibody ; TARGET ; LESIONS ; ASSAY ; GLUTATHIONE ; WOMEN ; cervical cancer ; CERVICAL-CANCER ; FUSION ; p53 ; human papillomavirus ; HPV ; E6 ; ONCOGENE ; HUMAN-PAPILLOMAVIRUS ; POSITIVE CANCER-CELLS ; PROTEIN-PROTEIN INTERACTIONS ; specificity ; glutathione-S-transferase ; E6 ONCOPROTEIN ; INHIBITORS ; CHEMISTRY ; LIBRARIES ; development ; methods ; USA ; COMPOUND ; microbiology ; premalignant lesions ; AlphaScreen((R)) ; E6-AP ; GST fusion protein ; papillornavirus
    Abstract: Human papillomavirus (HPV) infection is responsible for the development of cervical cancer and its premalignant lesions in women. The virus-encoded oncogene E6 is a promising target for an anti-HPV drug therapy. The authors describe the development of a homogenous screening assay for inhibitors of the E6 interaction with its cellular target, the E6-associated protein (E6AP), based on AlphaScreen (R) technology. The E6 protein was expressed and purified as glutathione S-transferase (GST) fusion protein, and the binding to a biotinylated E6AP peptide was monitored using GST-detecting Acceptor beads coated either with anti-GST antibody or glutathione. After optimization of the assay conditions, a commercial library of 3000 compounds was screened for inhibitors. Active compounds were retested and counterscreened for E6/E6AP specificity using biotinylated GST as a control protein. The results obtained with both types of GST-detecting reagents correlated very well and demonstrated the great potential of the newly developed glutathione-coated Acceptor beads as a detection reagent for GST fusion proteins
    Type of Publication: Journal article published
    PubMed ID: 17478484
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  • 4
    Keywords: Germany ; human ; DISEASE ; DISEASES ; HISTORY ; POPULATION ; DISTINCT ; PROTEIN ; TIME ; INFECTION ; SKIN ; papillomavirus ; ALPHA ; antibodies ; antibody ; PATTERNS ; AGE ; WOMEN ; MEN ; CAPSID PROTEIN ; human papillomavirus ; VIRUS-LIKE PARTICLES ; HIGH-RISK ; HPV ; BETA ; HUMAN-PAPILLOMAVIRUS ; Jun ; SQUAMOUS-CELL CARCINOMA ; POLYMERASE-CHAIN-REACTION ; L1 ; CHILDREN ; NATURAL-HISTORY ; PREVALENCE ; NONMELANOMA SKIN CANCERS ; glutathione-S-transferase ; SERUM ; ADULT ; ADULTS ; SAN-FRANCISCO ; review ; RE ; PATTERN ; papillomaviruses ; GAMMA ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; HPV 16 ; USA ; YOUNG-ADULTS ; INFECTIOUS-DISEASES ; microbiology ; serology ; multiplex serology ; SEROPREVALENCE ; GENERAL-POPULATION ; HPV types ; MAJOR CAPSID PROTEIN ; HPV-16 ; CUTANEOUS HUMAN PAPILLOMAVIRUSES ; FOOD-CONSUMPTION HABITS ; NORMAL CERVICAL SMEARS
    Abstract: The natural history of infections with many human papillomavirus (HPV) types is poorly understood. Here, we describe for the first time the age-and sex-dependent antibody prevalence for 29 cutaneous and five mucosal HPV types from 15 species within five phylogenetic genera (alpha, beta, gamma, mu, nu) in a general population. Sera from 1,797 German adults and children (758 males and 1,039 females) between 1 and 82 years (median 37 years) were analysed for antibodies to the major capsid protein L1 by Luminex-based multiplex serology. The first substantial HPV antibody reactions observed already in children and young adults are those to cutaneous types of the genera nu (HPV 41) and mu (HPV 1, 63). The antibody prevalence to mucosal high-risk types, most prominently HPV 16, was elevated after puberty in women but not in men and peaked between 25 and 34 years. Antibodies to beta and gamma papillomaviruses (PV) were rare in children and increased homogeneously with age, with prevalence peaks at 40 and 60 years in women and 50 and 70 years in men. Antibodies to cutaneous alpha PV showed a heterogeneous age distribution. In summary, these data suggest three major seroprevalence patterns for HPV of phylogenetically distinct genera: antibodies to mu and nu skin PV appear early in life, those to mucosal alpha PV in women after puberty, and antibodies to beta as well as to gamma skin PV accumulate later in life
    Type of Publication: Journal article published
    PubMed ID: 18566657
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  • 5
    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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  • 6
    Keywords: PEPTIDE ; INFECTION ; EFFICACY ; TYPE-16 ; VIRUS-LIKE PARTICLES ; MONOCLONAL-ANTIBODIES ; HPV16 ; VACCINE ; EPITOPE ; NEUTRALIZATION ; monoclonal antibody ; MINOR CAPSID PROTEIN ; pseudovirion ; prophylactic vaccine ; Immune epitope ; L2 protein
    Abstract: The N-terminal region of the human papillomavirus (HPV) L2 protein has been shown to contain immune epitopes able to induce the production of neutralizing and cross-neutralizing antibodies (Gambhira et al., 2007; Kawana et al., 1999). Using bacterial thioredoxin as a scaffold, we managed to enhance the immunogenicity of putative L2 neutralizing epitopes, but only a minor fraction of the resulting immune responses was found to be neutralizing (Rubio et al., 2009). To determine the recognition patterns for non-neutralizing, neutralizing and cross-neutralizing antibodies, we isolated and characterized a panel of 46 monoclonal antibodies directed against different HPV16 L2 epitopes. Four of such antibodies proved to be neutralizing, and two of them, both targeting the amino acid (aa) 20-38 region of L2, were found to cross-neutralize a broad range of papillomaviruses. The epitopes recognized by neutralizing and cross-neutralizing antibodies were mapped at high resolution and were found to be characterized by distinct recognition patterns. Even in the case of the L2 20-38 epitope, cross-neutralization of HPV31 pseudovirions proved to be extremely inefficient, and this was found to be primarily due to the lack of a proline residue at position 30. HPV16 specific amino acids in this region also appear to be responsible for the lack of cross-neutralizing activity, thus suggesting a potential immune escape mechanism. For the aa 71-80 region, instead, the data indicate that restriction of neutralization to HPV16 is due to sequence (or structural) differences laying outside of the epitope. Besides providing new insights on the molecular bases of L2-mediated immune reactivity, the present data may pave the way to novel vaccination approaches specifically evoking cross-neutralizing antibody responses.
    Type of Publication: Journal article published
    PubMed ID: 21074234
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  • 7
    Keywords: COHORT ; DISEASE ; HPV ; SAFETY ; UNITED-STATES ; YOUNG-WOMEN ; SEX-DIFFERENCES ; ANTIBODY-RESPONSE ; anal cancer ; AGED 18-45 YEARS
    Abstract: Background. We compared the immunogenicity and reactogenicity of Cervarix or Gardasil human papillomavirus (HPV) vaccines in adults infected with the human immunodeficiency virus (HIV). Methods. This was a double-blind, controlled trial randomizing HIV-positive adults to receive 3 doses of Cervarix or Gardasil at 0, 1.5, and 6 months. Immunogenicity was evaluated for up to 12 months. Neutralizing anti-HPV-16/18 antibodies were measured by pseudovirion-based neutralization assay. Laboratory tests and diary cards were used for safety assessment. The HPV-DNA status of the participants was determined before and after immunization. Results. Ninety-two participants were included in the study. Anti-HPV-18 antibody titers were higher in the Cervarix group compared with the Gardasil group at 7 and 12 months. No significant differences in anti-HPV-16 antibody titers were found among vaccine groups. Among Cervarix vaccinees, women had higher anti-HPV-16/18 antibody titers compared to men. No sex-specific differences in antibody titers were found in the Gardasil group. Mild injection site reactions were more common in the Cervarix group than in the Gardasil group (91.1% vs 69.6%; P = .02). No serious adverse events occurred. Conclusions. Both vaccines were immunogenic and well tolerated. Compared with Gardasil, Cervarix induced superior vaccine responses among HIV-infec
    Type of Publication: Journal article published
    PubMed ID: 24273179
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  • 8
    Abstract: Ninety-one HIV-infected individuals (61 men and 30 women) were randomized to vaccination either with quadrivalent (Gardasil) or bivalent (Cervarix) HPV vaccine. Neutralizing and specific HPV-binding serum antibodies were measured at baseline and 12 months after the first vaccine dose. Presence of neutralizing and binding antibodies had good agreement (average Kappa for HPV types 6, 11, 16, 18, 31, 33 and 45 was 0.65). At baseline, 88% of subjects had antibodies against at least one genital HPV. Following vaccination with Cervarix, all subjects became seropositive for HPV16 and 18. After Gardasil vaccination, 96% of subjects seroconverted for HPV16 and 73% for HPV18. Levels of HPV16-specific antibodies were 〈1 international unit (IU) in 87% of study subjects before vaccination but 〉10IU in 85% of study subjects after vaccination. Antibodies against non-vaccine HPV types appeared after Gardasil vaccination for 〉50% of vaccinated females for HPV 31, 35 and 73 and for 〉50% of Cervarix-vaccinated females for HPV 31, 33, 35, 45, 56 and 58. Cross-reactivity with non-genital HPV types was also detected. In conclusion, HIV-infected subjects responded to HPV vaccination with induction of neutralizing antibodies against both vaccine and non-vaccine types.
    Type of Publication: Journal article published
    PubMed ID: 26896686
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  • 9
    Keywords: PEPTIDE ; CELLS ; EXPRESSION ; IN-VITRO ; tumor ; TUMOR-CELLS ; Germany ; human ; PROTEIN ; PROTEINS ; TUMORS ; MICE ; RELEASE ; COMPLEX ; RESPONSES ; COMPLEXES ; SERA ; REDUCTION ; DENDRITIC CELLS ; BINDING ; SEQUENCE ; antibodies ; NEUTRALIZING ANTIBODIES ; PARTICLES ; ASSAY ; ESCHERICHIA-COLI ; GLUTATHIONE ; LYMPHOCYTES ; VIRUS-LIKE PARTICLES ; HPV ; HPV16 ; VACCINE ; IMMUNE-RESPONSE ; vaccination ; L1 ; SEQUENCE-ANALYSIS ; glutathione-S-transferase ; HPV PSEUDOVIRIONS ; NASAL IMMUNIZATION
    Abstract: We analyzed capsomeres of human papillomavirus type 16 (HPV16) consisting of the L1 major structural protein for their ability to trigger a cytotoxic T-cell (CTL) response. To this end, we immunized C57BL/6 mice and used the L1(165-173) peptide for ex vivo restimulation of splenocytes prior to analysis (Cr-51 release assay and enzyme-linked immunospot assay [ELISPOT]). This peptide was identified in this study as a D-b-restricted naturally processed CTL epitope by HPV16 L1 sequence analysis, major histocompatibility complex class I binding, and Cr-51 release assays following immunization of C57BL/6 mice with HPV16 L1 virus-like particles (VLPs). HPV16 L1 capsomeres were obtained by purification of HPV16 L1 lacking 10 N-terminal amino acids after expression in Escherichia coli as a glutathione S-transferase fusion protein (GST-HPV16 L1DeltaN10). Sedimentation analysis revealed that the majority of the purified protein consisted of pentameric capsomeres, and assembled particles were not observed in minor contaminating higher-molecular-weight material. Subcutaneous (s.c.) as well as intranasal immunization of C57BL/6 m ice with HPV16 L1 capsomeres triggered an L1-specific CTL response in a dose- dependent manner as measured by ELISPOT and Cr-51 release as say. Significant reduction of contaminating bacterial endotoxin (lipopolysaccharide) from the capsomere preparation did not diminish the immunogenicity. Antibody responses (serum and vaginal) were less robust under the experimental conditions employed. In addition, s.c. vaccination with HPV16 L1 capsomeres induced regression of established tumors expressing L1 determinants (C3 tumor cells). Our data demonstrate that capsomeres are potent inducers of CTL responses similar to completely assembled T=7 VLPs. This result is of potential relevance for the development of (combined prophylactic and therapeutic) HPV-specific vaccines, since capsomeres can be produced easily and also can be modified to incorporate heterologous sequences such as early HPV proteins
    Type of Publication: Journal article published
    PubMed ID: 12663770
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  • 10
    Keywords: CELLS ; RISK ; TISSUE ; DNA ; ASSOCIATION ; antibodies ; antibody ; PCR ; human papillomavirus ; HPV ; SQUAMOUS-CELL CARCINOMA ; HEAD ; L1 ; PREVALENCE ; POLYMERASE CHAIN-REACTION ; glutathione-S-transferase ; NECK-CANCER ; HUMAN-PAPILLOMAVIRUS HPV
    Abstract: According to PCR, the prevalences of human papillomavirus (HPV) DNA were 6.3% (13 of 206) in tonsillitis or hypertrophic tonsillar tissues and 0.6% (1 of 174) in exfoliated cells from normal tonsils. HPV-16 was the only type detected in tonsillar tissues, but it did not appear to lead to L1 antibody production
    Type of Publication: Journal article published
    PubMed ID: 15750119
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