| 1. |
- Bissett, Sara L., et al.
(author)
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Human Papillomavirus Antibody Reference Reagents for Use in Postvaccination Surveillance Serology
- 2012
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In: Clinical and Vaccine Immunology. - 1556-6811 .- 1556-679X. ; 19:3, s. 449-451
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Journal article (peer-reviewed)abstract
- Suitably controlled serosurveillance surveys are essential for evaluating human papillomavirus (HPV) immunization programs. A panel of plasma samples from 18-year-old females was assembled, the majority of the samples being from recipients of the bivalent HPV vaccine. Antibody specificities were evaluated by three independent laboratories, and 3 pools that displayed no antibodies to any HPV type tested or intermediate or high levels of antibody to HPV16, HPV18, HPV31, and HPV45 were created. These pools will be useful as control reagents for HPV serology.
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| 2. |
- Bzhalava, Davit, et al.
(author)
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Phylogenetically diverse TT virus viremia among pregnant women
- 2012
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In: Virology. - : Elsevier BV. - 1096-0341 .- 0042-6822. ; 432:2, s. 427-434
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Journal article (peer-reviewed)abstract
- Infections during pregnancy have been suggested to be involved in childhood leukemias. We used high-throughput sequencing to describe the viruses most readily detectable in serum samples of pregnant women. Serum DNA of 112 mothers to leukemic children was amplified using whole genome amplification. Sequencing identified one TT virus (TTV) isolate belonging to a known type and two putatively new TTVs. For 22 mothers, we also performed ITV amplification by general primer PCR before sequencing. This detected 39 TTVs, two of which were identical to the Tilts found after whole genome amplification. Altogether, we found 40 TTV isolates, 29 of which were putatively new types (similarities ranging from 89% to 69%). In conclusion, high throughput sequencing is useful to describe the known or unknown viruses that are present in serum samples of pregnant women. (C) 2012 Elsevier Inc. All rights reserved.
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| 3. |
- Bzhalava, Davit, et al.
(author)
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Unbiased Approach for Virus Detection in Skin Lesions
- 2013
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In: PLOS ONE. - : Public Library of Science. - 1932-6203. ; 8:6
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Journal article (peer-reviewed)abstract
- To assess presence of virus DNA in skin lesions, swab samples from 82 squamous cell carcinomas of the skin (SCCs), 60 actinic keratoses (AKs), paraffin-embedded biopsies from 28 SCCs and 72 kerathoacanthomas (KAs) and fresh-frozen biopsies from 92 KAs, 85 SCCs and 92 AKs were analyzed by high throughput sequencing (HTS) using 454 or Ion Torrent technology. We found total of 4,284 viral reads, out of which 4,168 were Human Papillomavirus (HPV)-related, belonging to 15 known (HPV8, HPV12, HPV20, HPV36, HPV38, HPV45, HPV57, HPV59, HPV104, HPV105, HPV107, HPV109, HPV124, HPV138, HPV147), four previously described putative (HPV 915 F 06 007 FD1, FA73, FA101, SE42) and two putatively new HPV types (SE46, SE47). SE42 was cloned, sequenced, designated as HPV155 and found to have 76% similarity to the most closely related known HPV type. In conclusion, an unbiased approach for viral DNA detection in skin tumors has found that, although some new putative HPVs were found, known HPV types constituted most of the viral DNA.
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| 4. |
- Faust, Helena, et al.
(author)
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Antibodies to merkel cell polyomavirus correlate to presence of viral DNA in the skin.
- 2011
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In: Journal of Infectious Diseases. - : Oxford University Press (OUP). - 1537-6613 .- 0022-1899. ; 203:8, s. 1096-1100
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Journal article (peer-reviewed)abstract
- To validate whether Merkel cell polyomavirus (MCV) serology correlates with MCV infection, we compared real-time polymerase chain reaction results for MCV DNA on fresh-frozen biopsy specimens from various skin lesions and healthy skin from 434 patients to MCV serology results using viruslike particles (VLPs) and MCV neutralization assays. Sixty-five percent of participants were MCV seropositive and 18% were MCV DNA positive. The presence of antibodies was correlated with the presence of virus DNA (odds ratio, 27.85 [95% confidence interval, 6.6-166.5]), with 97% of patients who tested positive for MCV DNA being MCV seropositive. VLP antibody levels correlated to neutralization titers (r = .72), and high antibody levels correlated to high MCV load (P < .01).
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| 5. |
- Faust, Helena, et al.
(author)
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Cutaneous Human Papillomaviruses and squamous cell carcinoma of the skin: Nested case-control study.
- 2016
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In: Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology. - 1538-7755.
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Journal article (peer-reviewed)abstract
- Cutaneous Human Papillomavirus (HPV) types have been associated with non-melanoma skin cancer (NMSC), including a previous nested case-control study using HPV serology with bacterially derived fusion proteins with the major HPV capsid protein L1 (GST-L1). However, HPV serology using conformationally intact pseudovirions has been shown to correlate better with natural infection. Prospective studies using a more valid marker of infection are therefore warranted.
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| 6. |
- Faust, Helena, et al.
(author)
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Human Papillomavirus neutralizing and cross-reactive antibodies induced in HIV-positive subjects after vaccination with quadrivalent and bivalent HPV vaccines.
- 2016
-
In: Vaccine. - : Elsevier BV. - 1873-2518 .- 0264-410X. ; , s. 1559-1559
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Journal article (peer-reviewed)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.
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| 7. |
- Faust, Helena, et al.
(author)
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Mutations in human papillomavirus type 16 L1 hypervariable surface-exposed loops affect L2 binding and DNA encapsidation
- 2013
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In: The Journal of general virology. - : Microbiology Society. - 1465-2099 .- 0022-1317. ; 94:Pt 8, s. 1841-1849
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Journal article (peer-reviewed)abstract
- Prophylactic vaccines against human papillomavirus (HPV) based on virus-like particles (VLP) induce type-specific neutralizing antibodies against a small number of hypervariable residues positioned in surface-exposed loops of the major capsid protein L1. To investigate the importance of these residues for neutralization, cross-neutralization, L2 incorporation and genome encapsidation, ten surface-exposed amino acid residues in four hypervariable loops of L1 were mutated. VLPs containing mutated or WT L1, with or without WT L2, were produced in 293TT cells using pseudovirion expression vectors. The mutations reduced the ability to induce neutralizing antibodies and to incorporate the L2 protein in the capsid. Ability to induce cross-neutralizing antibodies and to encapsidate pseudogenomes were completely abrogated. In summary, the surface-exposed L1 loops are important for the function of the HPV particle.
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| 8. |
- Faust, Helena, et al.
(author)
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Prevalence of human papillomavirus types, viral load and physical status of HPV16 in head and neck squamous cell carcinoma from the South Swedish Health Care Region
- 2016
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In: Journal of General Virology. - : Microbiology Society. - 0022-1317 .- 1465-2099. ; 97:11, s. 2949-2956
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Journal article (peer-reviewed)abstract
- Incidence of human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) is rising in several countries. Intriguingly, large variations of HPV16 viral load and different proportions of the physical viral status among HNSCC have been reported. We analysed fresh biopsies of 275 HNSCC patients from the South Swedish Health Care Region for HPV types with modified general primer PCR and Luminex. Seventy-eight HPV16-positive HNSCC cases were further investigated for viral DNA load and physical status using quantitative PCR for HPV E2 and E7 genes. Presence of intact E2 gene, as a surrogate marker for episomal HPV, was investigated with conventional PCR. Fifteen different HPV types were detected in HNSCC cases and HPV16 was present in 74% of the HPV-positive cases. HPV was detected in 65% (92/141) and 11% (15/134) of oropharyngeal and non-oropharyngeal carcinomas, respectively (P<0.0001). HPV was detected in 73% (75/103) of tonsillar carcinomas. The median load of HPV16 was 13 copies cell-1 (range 0.003–1080). Among HPV16-positive patients with oropharyngeal carcinoma, metastases to regional lymph nodes were observed in 100% (17/17) and 68% (40/58) for those with <1 HPV16 copy cell-1 and >1 HPV16 copy Cell-1, respectively (P=0.007). Among HPV16 cases, purely integrated HPV16 was found in 6%, whereas entirely episomal and mixed virus was detected in 51 and 42% of cases, respectively. Conclusively, HPV16 viral DNA load demonstrated a large diversity among HNSCCs. Although integration of HPV16 is common (48%), the episomal HPV16 is salient (93%) among HPV16 HNSCCs. In addition, low amount of HPV16 was associated with lymph node metastases among oropharyngeal carcinomas.
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| 9. |
- Faust, Helena, et al.
(author)
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Prospective study of merkel cell polyomavirus and risk of merkel cell carcinoma.
- 2014
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In: International Journal of Cancer. - : Wiley. - 0020-7136 .- 1097-0215. ; 134:4, s. 844-848
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Journal article (peer-reviewed)abstract
- Merkel cell carcinoma (MCC) is a rare type of skin cancer that has a characteristically increased incidence among immunosuppressed subjects. The DNA of Merkel cell polyomavirus (MCV) is regularly found in most MCC tumors. We investigated whether Merkel cell polyomavirus (MCV) infection increases the risk for future Merkel cell carcinoma (MCC). Two large biobank cohorts (Southern Sweden Microbiology Biobank and the Janus Biobank), containing samples from 856,000 healthy donors, were linked to the Cancer Registries in Sweden and Norway to identify cases of MCC occurring up to 30 years after donation of a serum sample. For each of the 22 cases (9 males and 13 females), four matched controls were included. The serum samples were analysed with an MCV neutralization assay and for IgG antibodies to MCV pseudovirions, using JC polyomavirus and cutaneous Human papillomaviruses as control antigens. An increased risk for future MCC was associated both with high levels of MCV antibodies (OR 4.4, 95% CI 1.3-17.4) and with MCV neutralizing activity (OR 5.3, 95% CI 1.3-32.3). In males, MCV seropositivity was not associated to MCC risk, whereas the risk was strongly increased in females, both for high levels of MCV antibodies (OR 7.0, 95% CI 1.6-42.8) and for MCV neutralizing activity (OR 14.3, 95% CI 1.7-677). In conclusion, we found prospective evidence that MCV infection is associated with an increased risk for future MCC, in particular among females. © 2013 Wiley Periodicals, Inc.
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| 10. |
- Faust, Helena, et al.
(author)
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Pseudovirion-binding and neutralizing antibodies to cutaneous Human Papillomaviruses correlated to presence of HPV DNA in skin.
- 2013
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In: Journal of General Virology. - : Microbiology Society. - 1465-2099 .- 0022-1317. ; 94, s. 1096-1103
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Journal article (peer-reviewed)abstract
- We compared seroreactivity to Human papillomavirus (HPV) antigens measured with two different high-throughput assays. One method used GST-L1 fusion proteins and the other heparin-bound HPV pseudovirions as antigens and both methods used multiplexed fluorescent beads for detection. For six HPV types (5, 6, 15, 16, 32 and 38), seroreactivity could be measured in parallel for 434 serum samples from non-immunosuppressed patients with skin lesions (squamous cell carcinoma of the skin, basal cell carcinoma of the skin, actinic keratosis and benign skin lesions). Biopsies from the skin lesions were tested for presence of HPV DNA using three different PCR methods, with typing by sequencing. Among the types included in the serological tests, HPV DNA of types HPV5, 15, 38 and 76 were most frequently detected in the tumours. Serum samples from subjects with HPV DNA positive biopsies and randomly selected serum samples from subjects with HPV DNA negative biopsies were also tested with neutralization assays with HPV5, 38 and 76 pseudovirions. Agreement of the three serological methods varied from poor to moderate and showed limited consistency. Type-specific seroprevalences among patients positive for the same type of HPV DNA (sensitivity of serology) was improved with the pseudovirion-based method (average of 40%, maximum 63%) compared to the GST-L1 method (average of 20%, maximum of 25%). Neutralization was the most sensitive assay for HPV38 (50%). In summary, the pseudovirion-based methods appeared to have an improved sensitivity.
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| 11. |
- Faust, Helena
(author)
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Pseudovirions in the Study of Papilloma- and Polyomaviruses
- 2012
-
Doctoral thesis (other academic/artistic)abstract
- The papilloma- and polyomaviruses are small DNA viruses that infect humans. Some members of these virus families can cause cancer in experimental animals and some are also established as carcinogenic to humans. Detection of specific antibodies to these viruses allows tracking past and present infections to elucidate natural history and association of infection with subsequent disease. As there are more than 150 types of Human papillomaviruses (HPV) and at least 10 Human polyomaviruses (HPyV), the study of antibodies to these virus families require high-throughput methods. Although it is difficult to produce infectious virus stocks of these viruses, so-called pseudovirions that morphologically and immunologically resemble native virions but lack the viral genome can be produced in vitro. In the present thesis, we produced such pseudovirions and used them to i) delineate the importance of hypervariable surface loops for the antigenicity and biological function of the HPV particle ii) developed and validate serological assays for measuring specific antibodies to HPV and Merkel cell polyomavirus (MCPyV) and iii) perform prospective seroepidemio-logical studies to evaluate whether infection with MCPyV was associated with an increased risk for Merkel cell carcinoma. Site-directed mutagenesis of the surface loops of the HPV capsid found that these loops were essential for the incorporation of the minor capsid protein L2, the genome encapsidation and proper immunogenicity of the particle. Pseudovirion-based methods were correlated to presence of viral DNA. The pseudovirion neutralization assays and multiplexed assays using pseudovirions bound to heparin-coated fluorescent beads for 21 HPV and 2 HPyV types were correlated with viral DNA for 16 HPV types and MCPyV. MCPyV specific antibody levels in serum were found to be strongly correlated to the MCPyV viral load in skin. Finally, biobank-based seroepidemiological studies found that MCPyV infection was associated with an increased risk for Merkel cell carcinoma (MCC), in particular among females.
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| 12. |
- Faust, Helena, et al.
(author)
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Serum antibodies to human papillomavirus (HPV) pseudovirions correlate with natural infection for 13 genital HPV types.
- 2013
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In: Journal of Clinical Virology. - : Elsevier BV. - 1386-6532. ; 56:4, s. 336-341
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Journal article (peer-reviewed)abstract
- BACKGROUND: Serology for human papillomaviruses (HPV) types -16 and -18 is established as an important tool for studies of HPV vaccinology and epidemiology. However, as there are a large number of oncogenic genital types of HPV there is a need for development of high-throughput, validated HPV serological assays that can be used for more comprehensive seroepidemiological studies and for research on multivalent HPV vaccines. OBJECTIVES: To develop a multiplexed pseudovirion-based serological assay (PsV-Luminex) encompassing 21 HPV types and validate the method by correlating the serology with the presence of type specific HPV DNA in cervical samples. STUDY DESIGN: Cervical swabs from 3,291 unvaccinated women attending organized cervical screening in Slovenia were tested with 3 different HPV DNA detection methods and presence of HPV DNA compared to presence of serum antibodies to pseudovirions from 15 genital HPV types (HPV-6,-11,-16,-18,-31,-33,-35,-39,-45,-52,-56,-58,-59,-68,-73). RESULTS: On average 51% of the HPV DNA positive women were seropositive for the same HPV type that was detected in the cervical specimen. We found a strong correlation with presence of HPV DNA and antibodies to the same HPV type for 13/15 genital HPV types (median OR=5.7, CI 95%=2.4-12.9). HPV-52 serology failed the validation and HPV-11 serology could not be validated because only a single woman was positive for HPV-11 DNA. The correlation between serology and HPV DNA status tended to be stronger among women infected with single HPV type (median OR=10.5, CI 95%=2.4-48.4) than among women with multiple HPV infections (median OR=4.6, CI 95%=1.8-11.7). CONCLUSIONS: A multiplexed HPV PsV-Luminex assay has been developed and validated to correlate with natural HPV infection for 13 HPV types, thus enabling more comprehensive studies in HPV epidemiology and vaccine research.
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| 13. |
- Faust, Helena, et al.
(author)
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Validation of multiplexed human papillomavirus serology using pseudovirions bound to heparin coated beads.
- 2010
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In: Journal of General Virology. - : Microbiology Society. - 1465-2099 .- 0022-1317. ; 91, s. 1840-1848
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Journal article (peer-reviewed)abstract
- We developed and validated a high-throughput human papillomavirus (HPV) serology method based on Luminex technology, using pseudovirions (PsVs) of eight mucosal HPV types (HPV 6, 11, 16, 18, 31, 45, 52 and 58) and two cutaneous HPV types (HPV 5 and 38) bound to heparin coated beads. Analysis with neutralising type-specific monoclonal antibodies against included HPV types indicated type-specificity of the assay. Analysis of negative control serum samples from 63 children and 71 middle-aged women with up to one lifetime sexual partner indicated high specificity. Positive control serum samples from subjects with known HPV DNA status or clinical diagnosis found expected sensitivities for most of the HPV types in 219 European serum samples, but less than expected in 124 samples from Africa. HPV 45 and 52 did not react as expected with the human serum samples. The Pseudovirion-Luminex method was used to determine the HPV-seropositivity-associated relative risk for future cervical cancer using 208 serum samples from a prospective study of 18814 women followed for 23 years, previously analysed with standard HPV 16 ELISA. The Pseudovirion-Luminex method gave similar results as ELISA (Kappa= 0.77). As expected, HPV seropositivities assayed using the Pseudovirion-Luminex method found increased risk for cervical cancer for HPV 16 (OR=7.7, CI 95%=2.6-23) and HPV 31(OR=4.1, CI 95%=1.6-10.8), non-significant tendencies for increased risk for other mucosal HPV types and no risk for the cutaneous HPV types. In summary, multiplexed HPV serology using mammalian-derived pseudovirions selected for native conformation by binding to heparin-coated beads is validated as a high-throughput HPV serological method, for most of the analysed HPV types.
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| 14. |
- Letsolo, Boitelo T, et al.
(author)
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Establishment and characterization of a human papillomavirus type 16-positive tonsillar carcinoma xenograft in BALB/c nude mice
- 2016
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In: Head and Neck. - : Wiley. - 1043-3074. ; 38:3, s. 25-417
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Journal article (peer-reviewed)abstract
- BACKGROUND: Among head and neck cancers, human papillomavirus type 16 (HPV16) is associated with tonsillar carcinomas. Despite this, no HPV16-positive tonsillar cancer cell line has been established in nude mice.METHODS: Fresh tonsillar carcinoma biopsies were obtained from 23 patients and implanted subcutaneously into nude mice (BALB/c, nu/nu).RESULTS: After 7 months, one xenograft was established. The primary tumor harbored 2.7 copies (95% confidence interval = 2.4-2.9) of HPV16/cell and displayed 99.9% (7904/7906) nucleotide identity to HPV16 (EU118173.1). The xenograft showed increased methylation in two E2-binding sites of the HPV16 genome. Both episomal and integrated HPV16 were detected in the original tumor and in 14 xenografts from the second passage. From this passage, a viral load of 6.4 copies/cell (range = 4.6-9.6) and 3.7 (range = 1.0-5.5) E7-mRNA transcripts/HPV16-genome were detected.CONCLUSION: This xenograft represents the first established HPV16-positive tonsillar tumor in nude mice and could provide an experimental system of HPV16-positive tonsillar cancers. © 2015 Wiley Periodicals, Inc. Head Neck 38: 417-425, 2016.
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| 15. |
- Schellenbacher, Christina, et al.
(author)
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Efficacy of RG1-VLP Vaccination against Infections with Genital and Cutaneous Human Papillomaviruses
- 2013
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In: Journal of Investigative Dermatology. - : Elsevier BV. - 1523-1747 .- 0022-202X. ; 133:12, s. 2706-2713
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Journal article (peer-reviewed)abstract
- Licensed human papillomavirus (HPV) vaccines, based on virus-like particles (VLPs) self-assembled from major capsid protein L1, afford type-restricted protection against HPV types 16/18/6/11 (or 16/18 for the bivalent vaccine), which cause 70% of cervical cancers (CxCas) and 90% of genital warts. However, they do not protect against less prevalent high-risk (HR) types causing 30% of CxCa, or cutaneous HPV. In contrast, vaccination with the minor capsid protein L2 induces low-level immunity to type-common epitopes. Chimeric RG1-VLP presenting HPV16 L2 amino acids 17-36 (RG1 epitope) within the DE-surface loop of HPV16 L1 induced cross-neutralizing antisera. We hypothesized that RG1-VLP vaccination protects against a large spectrum of mucosal and cutaneous HPV infections in vivo. Immunization with RG1-VLP adjuvanted with human-applicable alum-MPL (aluminum hydroxide plus 3-O-desacyl-4'-monophosphoryl lipid A) induced robust L2 antibodies (ELISA titers 2,500-12,500), which (cross-)neutralized mucosal HR HPV16/18/45/37/33/52/58/35/39/51/59/68/73/26/69/34/70, low-risk HPV6/11/32/40, and cutaneous HPV2/27/3/76 (titers 25-1,000) using native virion-or pseudovirion (PsV)-based assays, and a vigorous cytotoxic T lymphocyte response by enzyme-linked immunospot. In vivo, mice were efficiently protected against experimental vaginal challenge with mucosal HR PsV types HPV16/18/45/31/33/52/58/35/39/51/59/68/56/73/26/53/66/34 and low-risk HPV6/43/44. Enduring protection was demonstrated 1 year after vaccination. RG1-VLP is a promising next-generation vaccine with broad efficacy against all relevant mucosal and also cutaneous HPV types.
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| 16. |
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| 17. |
- Ucakar, Veronika, et al.
(author)
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Pre-vaccination seroprevalence of 15 human papillomavirus (HPV) types among women in the population-based Slovenian cervical screening program
- 2013
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In: Vaccine. - : Elsevier BV. - 1873-2518 .- 0264-410X. ; 31:43, s. 4935-4939
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Journal article (peer-reviewed)abstract
- Objectives: To estimate seroprevalence of 11 high-risk (hr) HPV types and four low-risk (lr) HPV types among 20-64 years old Slovenian women participating in the population-based cervical cancer screening program. Methods: Serum samples from 3259 women were tested for HPV type-specific antibodies with a multiplexed pseudovirion-based serological assay (PsV-Luminex). Results: Seropositivity for any of the 15 HPV types was 65.7%, any of the 11 hr-HPV types 59.2%, and any of the four lr-HPV types 33.1%. Antibodies against at least one of the four vaccine HPV types (HPV 6, 11, 16, 18) were detected in 40.8% women. Among hr-HPV types, seropositivity was highest for HPV 16 (25.2%) and among lr-HPV types for HPV 6(19.1%). Age-specific HPV16 seropositivity was highest among 30-39 years old (29.6%) and decreased with increasing age to 14.0% among 60-64 years old. Conclusion: The lifetime sexual exposure to genital HPV types is substantial, emphasising the need for HPV vaccination. (C) 2013 The Authors. Published by Elsevier Ltd. All rights reserved.
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