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Background: Alpha-human papillomavirus (α-HPV) plays a causal role in cervical cancer, but little is known about the epidemiology of genital Beta-human papillomavirus (β-HPV) infection.Methods: We used Luminex and PCR hybridization to detect β- and α-HPVs prevalence at enrollment and 12-month follow-up in cervical samples from 505 women enrolled in the Ludwig-McGill cohort study. We compared epidemiologic correlates of both β- and α-HPVs and compared genotypes between these genera with respect to co-occurrence and association with cervical cytologic abnormalities.Results: Infection with β-HPV types was more prevalent than that with α-HPV types at both visits (cumulative prevalences: 27.3% vs. 21.6%, respectively, P = 0.034). β-HPVs were mostly transient; however, only 1.98% women retained their original positivity at 12 months, whereas persistence was higher for α-HPVs (5.15%; P = 0.007). Age, parity, and sexual activity variables were predictors of α-HPV but not of β-HPV. α- and β-HPV types occurred independently. Increased risk of cervical abnormalities was restricted to women infected with α-9 or α-6 HPV types. We found no epidemiologic correlates for β-HPV infections.Conclusions: Detection of β-HPV types in the cervix tends to occur as random and transient episodes not explained via the sexual-transmission correlates that characterize infections by α-HPVs.Impact: Although it is plausible that β-HPVs may play a direct or indirect carcinogenic role, the lack of epidemiologic correlates for detection episodes of these viruses and lack of association with cervical lesions speak against their ancillary role as sexually transmitted agents in cervical carcinogenesis. Cancer Epidemiol Biomarkers Prev; 26(8); 1312–20. ©2017 AACR.

Supplementary Table S1. Positivity at both enrollment and follow-up for selected HPV types in the Ludwig-McGill Cohort Study (n=503)

Supplementary Figure S1. Selection of participants from the Ludwig-McGill Cohort Study for inclusion in the present study. Enrollment and 12-month samples and data were included to permit the analysis of one-year infection persistence.

Human papillomavirus (HPV)-driven head/neck squamous cell carcinomas (HNSCC) prevalence varies globally. We evaluated HPV DNA and p16HPV was genotyped by PCR-hybridization. All HPV DNA positive and some HPV DNA negative cases underwent p16HPV DNA was detected in 32.8%, 11.1%, and 17.8% of oropharyngeal (OPC), oral cavity (OCC) and laryngeal (LC) cancers, respectively. OPC HPV prevalence was higher in Colombia (94.7%), and Argentina (42.6%) compared to Brazil (10.6%) and Peru (0.0%). HPV-16 was the most detected. Other HPVs were found in LC. Higher rates of p16Our results support a role for HPV-16 in a subset of HNSCC, corroborate the heterogeneity observed in samples from different countries, and contribute additional etiological and biomarkers information in tumors of significant impact worldwide.

It is suggested that HPV-18 variants from the A lineage have higher oncogenic potential compared to B variants. Some studies show uneven distribution of HPV-18 variants in cervical adenocarcinomas and squamous cell carcinomas. Regarding HPV-18 variants’ functions, the few studies reported focus on E6, and none were performed using natural host cells. Here, we immortalized primary human keratinocytes (PHKs) with E6/E7 of HPV-18 A1 and B1 sublineages and functionally characterized these cells. PHK18A1 reached immortalization significantly faster than PHK18B1 and formed a higher number of colonies in monolayer and 3D cultures. Moreover, PHK18A1 showed greater invasion ability and higher resistance to apoptosis induced by actinomycin-D. Nevertheless, no differences were observed regarding morphology, proliferation after immortalization, migration, or epithelial development in raft cultures. Noteworthy, our study highlights qualitative differences among HPV-18 A1 and B1 immortalized PHKs: in contrast to PHK18A1, which formed more compact colonies and spheroids of firmly grouped cells and tended to invade and migrate as clustered cells, morphologically, PHK18B1 colonies and spheroids were looser, and migration and invasion of single cells were observed. Although these observations may be relevant for the association of these variants with cervical cancer of different histological subtypes, further studies are warranted to elucidate the mechanisms behind these findings.

As funcoes das oncoproteinas E6 e E7 do Papilomavirus Humano tipo 16 (HPV-16) na progressao de celulas epiteliais imortalizadas para tumores invasivos nao sao totalmente compreendidas. Aqui, estabelecemos uma nova ligacao entre E6 e E7 de duas variantes moleculares de HPV-16 (AA e E-350G) e c-MYC, em relacao a cooperacao na promocao da transformacao maligna de queratinocitos humano primarios de prepucio de recem-nascido (PHK). Nosso objetivo foi estudar os efeitos sinergicos de E6/E7 e c-MYC na proliferacao e no potencial de transformacao in vitro de PHKs. Avaliou-se a proliferacao celular atraves da expressao proteica do Antigeno Nuclear de Celulas Proliferantes (PCNA). Tambem avaliamos a capacidade de transformacao, in vitro, dos PHKs atraves de dois ensaios complementares. Observamos que E-350G-c-MYC PHKs exibiram um aumento discreto na expressao de PCNA e formaram significativamente mais colonias tanto nos ensaios de soft-agar quanto nos ensaios em placas de cultura de baixa adesao. No geral, concluimos que a variante E-350G co-transfectada com c-MYC pode promover a transformacao celular maligna com eficiencia maior do que a variante AA-c-MYC. As propriedades oncogenicas exibidas pela variante E-350G permitem entender em maior detalhe os mecanismos que podem levar a neoplasia cervical humana, dada a maior frequencia de sua ocorrencia na progressao de lesoes precursoras de alto grau para carcinomas invasivos.

HPV-16 causes approximately 90% of anal canal (AC) cancers worldwide. This study aimed to evaluate the prevalence and persistence of HPV-16 genetic variants in the AC of men from three different countries (Brazil, Mexico and United States) and to further identify sociodemographic and behavioral factors associated with these infections.Participants from the multinational prospective HPV Infection in Men (HIM) Study who had at least one HPV-16 positive AC swab were included. Characterization into HPV-16 genetic variants was successfully performed by PCR-sequencing in 95.6% (217/227) samples and these were classified into HPV-16 lineages and sublineages.We observed higher prevalence of lineage A variants, mainly from A1 sublineage, in all countries. Non-A lineage variants were mostly detected in men from Brazil, where higher diversity of sublineage variants was detected during follow-up. Compare to men detected with Non-A HPV-16 lineage variants, men infected with lineage A reported a higher lifetime number of female sexual partners. Finally, a significantly higher prevalence of Non-A lineage variants was observed among men who have sex with men (MSM) with a transient HPV-16 AC infection (p = 0.033), but no significant differences regarding variants lineages and persistence status were observed when stratified by country, self-reported ethnicity or age.Our data extend previous reports which indicate that globally HPV-16 variants are unevenly distributed, and contribute further to studies of the natural history of AC HPV infections in men.

Made available in DSpace on 2021-06-25T11:02:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Human papillomavirus (HPV) types 6 and 11 are the etiological agents of recurrent respiratory papillomatosis (RRP). We examined the prevalence and distribution of HPVs 6 and 11 genetic variants in juvenile onset (JORRP) and adult onset (AORRP) laryngeal papillomas. Cases of JORRP and AORRP were collected, retrospectively. HPV detection and genotyping were accessed by polymerase chain reaction-sequencing in 67 RRP samples. Overall, the most prevalent HPV-6 variants were from B1 (55.8%) and B3 (27.9%) sublineages, whereas among HPV-11 positive samples A2 (62.5%) variants were predominant. A higher prevalence of HPV-6 B1 was observed in JORRP (83.3% B1 and 16.7% B3), compared with AORRP cases (58.3% B1 and 41.7% B3). HPV-11 A2 variants were more prevalent both in JORRP (57.2%) and in AORRP cases (70.0%). Nevertheless, with the exception that HPV-6 B1 were significantly less likely to recur, there was a lack of association between any particular HPVs 6 or 11 variant and clinicopathological features. Our data do not support an association between HPVs 6 and 11 variability and RRP. Center for Translational Research in Oncology Instituto do Cancer do Estado de Sao Paulo Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo Department of Bioscience and Oral Diagnosis São Paulo State University (UNESP) Centro Clínico de Cabeza y Cuello Department of Pathology and Forensic Medicine Ribeirao Preto Medical School Universidade de São Paulo Department of Otorhinolaryngology and Head and Neck Surgery Sao Paulo Federal University Department of Radiology and Oncology Faculdade de Medicina Universidade de São Paulo Department of Bioscience and Oral Diagnosis São Paulo State University (UNESP) FAPESP: 16/24019-0 FAPESP: 17/23211-8 CNPq: 303431/2018-0 CNPq: 306326/2015-9

Background: Alpha-human papillomavirus (α-HPV) plays a causal role in cervical cancer, but little is known about the epidemiology of genital Beta-human papillomavirus (β-HPV) infection. Methods: We used Luminex and PCR hybridization to detect β- and α-HPVs prevalence at enrollment and 12-month follow-up in cervical samples from 505 women enrolled in the Ludwig-McGill cohort study. We compared epidemiologic correlates of both β- and α-HPVs and compared genotypes between these genera with respect to co-occurrence and association with cervical cytologic abnormalities. Results: Infection with β-HPV types was more prevalent than that with α-HPV types at both visits (cumulative prevalences: 27.3% vs. 21.6%, respectively, P = 0.034). β-HPVs were mostly transient; however, only 1.98% women retained their original positivity at 12 months, whereas persistence was higher for α-HPVs (5.15%; P = 0.007). Age, parity, and sexual activity variables were predictors of α-HPV but not of β-HPV. α- and β-HPV types occurred independently. Increased risk of cervical abnormalities was restricted to women infected with α-9 or α-6 HPV types. We found no epidemiologic correlates for β-HPV infections. Conclusions: Detection of β-HPV types in the cervix tends to occur as random and transient episodes not explained via the sexual-transmission correlates that characterize infections by α-HPVs. Impact: Although it is plausible that β-HPVs may play a direct or indirect carcinogenic role, the lack of epidemiologic correlates for detection episodes of these viruses and lack of association with cervical lesions speak against their ancillary role as sexually transmitted agents in cervical carcinogenesis. Cancer Epidemiol Biomarkers Prev; 26(8); 1312–20. ©2017 AACR.

Background: Human papillomavirus (HPV) types 6 and 11 are mainly associated with the development of genital warts and recurrent respiratory papillomatosis. We examined intratypic genetic variability of both viral types with the development of cervical cytologic abnormalities in Brazilian women. Methods: We used PCR sequencing to characterize variants of HPVs 6 and/or 11 in cervical swabs from women in the Ludwig–McGill Cohort Study. We used a binomial generalized estimating equations (GEE) model with logit link to estimate odds ratios (OR) and 95% confidence intervals (CI) for the associations between HPV 6 and 11 variants and cytologic abnormalities. Results: B1 and B3 HPV6 and A2 HPV11 variants were the most common isolates identified. Compared with HPV6-negative women, the ORs among women harboring HPV6 B1 or B3 variants were 6.3 (95% CI, 2.3–17.0) and 2.3 (95% CI, 0.6–9.7) for atypical cells of undetermined significance (ASCUS)/low squamous intraepithelial lesions (LSIL), respectively, and 1.7 (95% CI, 0.6–5.1) and 1.2 (95% CI, 0.3–4.7) for ASCUS/LSIL/high squamous intraepithelial lesions (HSIL). Respective ORs were 5.0 (95% CI, 1.7–14.6) and 2.8 (95% CI, 1.0–8.1) upon comparing women with HPV11 A2 variants to HPV11-negative women. All associations disappeared when adjusting for coinfections with high-risk HPV types. Conclusions: Our data do not support an association between low-risk HPVs 6 and 11 genetic variability and cervical abnormalities. Impact: Risk of cervical cytologic abnormalities is not affected by intratypic polymorphism in HPVs 6 and 11.

Made available in DSpace on 2020-12-12T01:51:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Objective: The objective was to investigate the prevalence of the Epstein-Barr virus (EBV) and its association with human papilloma virus (HPV) detection, clinicopathological features, and the severity of recurrent respiratory papillomatosis (RRP). Methods: Cases of juvenile recurrent respiratory papillomatosis (JRRP) (n = 36) and adult recurrent respiratory papillomatosis (ARRP) (n = 44) were collected retrospectively and subdivided into low- and high-risk severity groups based on the Derkay score. We performed HPV detection and genotyping using a reverse hybridization protocol and investigated the presence of EBV by polymerase chain reaction (PCR) and in situ hybridization. CD21 levels were accessed by immunohistochemistry. Results: All samples were HPV-positive, including 49 cases of HPV 6, 26 cases of HPV 11, four cases of HPV 6 and 11 coinfections, and one case of HPV 16. EBV-DNA was detected in nine samples by PCR, although none of the cases were positive by means of in situ hybridization. CD21 immunoexpression was not statistically associated with any of the variables analyzed. HPV 6 detection was significantly higher in ARRP cases (P = 0.03), whereas HPV 11 was more prevalent in JRRP cases (P = 0.02) and was even more prevalent in JRRP cases of greater severity (Derkay laryngoscopic scale ≥20) (P = 0.04). Conclusion: The presence of EBV does not seem to play an important role in the progression/severity of RRP. Level of Evidence: 4 Laryngoscope, 2019. Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (Unesp), São José dos Campos Department of Stomatology Collective Health and Legal Dentistry Ribeirão Preto School of Dentistry Department of Pathology and Forensic Medicine Ribeirão Preto Medical School University of São Paulo Ribeirão Preto Department of Otolaryngology and Head and Neck Surgery Federal University of Sao Paulo Center for Translational Investigation in Oncology Instituto do Cancer do Estado de Sao Paulo Hospital das Clinicas da Faculdade de Medicina da Universidade de São Paulo Department of Radiology and Oncology Faculdade de Medicina Universidade de São Paulo Centro Clínico de Cabeza y Cuello Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (Unesp), São José dos Campos

The expression profile of angiogenesis associated proteins was assessed in patients with locally advanced cervical cancer included in a prospective, randomized phase II study. The majority of patients had squamous cell carcinoma (87.8%). Group A individuals were treated with neoadjuvant chemotherapy (cisplatin and gemcitabine) (NAC), followed by chemoradiation with cisplatin (CRT). Group B patients received CRT. Serum samples from patients in group A were collected at 03 different time points (at baseline (n=34), after NAC (n=24), and after CRT (n=27)). Samples of group B were collected in 02 different time points (at baseline (n=40) and after CRT (n=34)). Samples were grouped and evaluated with a non-parametric Mann-Whitney test where we considered the points as independent groups. The Friedman test (ANOVA) was used for analysis of group A (with three evaluation points). The levels of different proteins associated with angiogenesis (EGF, angiopoietin, G-CSF, endothelin-1, FGF-1, follistatin, IL-8, HGF, HB-EGF, PLGF, VEGF- D, VEGF-A and FGF-2) were assessed by a Multiplex Assay using the Luminex® technology. HB-EGF, IL-8 and VEGF-C were significantly decreased in patients from group A, whereas all others biomarkers did not present intra or intergroup differences. NAC was associated with an inferior PFS (progression free survival), with 3-year PFS rates of 40.9% compared with 60.4% in the CRT arm (HR, 1.84; 95% CI, 1.04 – 3.26; P = .033). NAC was also associated with a lower OS (overall survival) (3-year OS, 60.7% vs 86.8%; HR, 2.79; 95% CI, 1.29-6.01; P=0.006). After treatment completion, complete response rates were 56.3% in the NAC arm and 80.3% in the CRT arm (P=0.008). Toxicities were similar in both arms, with the exception of hypomagnesemia and neuropathy that were more commonly observed within NAC individual. We can conclude, within the limitations of presented study, that the NAC in group A results in significant alterations in specific markers associated with angiogenesis. Citation Format: ROBERTO JUN ARAI, THIAGO RODRIGUES DA COSTA, SILVANEIDE FERREIRA, LAURA SICHERO, MARIA PILAR ESTEVEZ DIZ. Assessment of angiogenesis markers in advanced cervical carcinoma patients that received chemoradiation therapy plus/minus neoadjuvant chemotherapy [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A038. doi:10.1158/1535-7163.TARG-19-A038

Human papillomavirus (HPV) infections are associated with the development of anogenital lesions in men. There are no reports describing the distribution of non-α HPV types in the anal canal of a sexually diverse group of men. The HPV Infection in Men (HIM) Study is a multicentre study on the natural history of HPV infection in Brazil, Mexico, and the USA. At baseline, 12% of anal canal PCR HPV-positive specimens were not typed by the Roche Linear Array, and were considered to be unclassified. Our goals were to characterize HPVs among these unclassified specimens at baseline, and to assess associations with participant socio-demographic and behavioural characteristics. Unclassified HPVs were typed by sequencing of amplified PGMY09/11 products or cloning of PGMY/GP + nested amplicons followed by sequencing. Further analysis was conducted with FAP primers. Of men with unclassified HPV in the anal canal, most (89.1%) were men who have sex with women. Readable sequences were produced for 62.8% of unclassified specimens, of which 75.2% were characterized HPV types. Eighteen, 26 and three different α-HPV, β-HPV and γ-HPV types were detected, respectively. α-HPVs were more commonly detected among young men (18–30 years) than among older men (45–70 years), whereas β-HPVs were more frequent among mid-adult men (31–44 years). β-HPVs were more common among heterosexual men (85.0%) than among non-heterosexual men. All β-HPVs detected among non-heterosexual men were β2-HPV types. The high prevalence of β-HPV in the anal canal of men who do not report receptive anal sex is suggestive of other forms of transmission that do not involve penile–anal intercourse.

The first publication that associated Human Papillomavirus (HPV) infection and esophageal cancer was published in 1982. However, data are still contradictory and require further investigation. The aim of this study was to identify high risk HPV DNA in esophageal tissue of patients with and without esophageal squamous cell carcinoma (ESCC) and correlate HPV presence with classical risk factors.Invited patients signed the informed consent form, and interviews were conducted in order to obtain information about sociodemographic and lifestyle behavior. During endoscopy, esophageal biopsies were collected from case and controls. Multiplex polymerase chain reaction genotyping was conducted on endoscopic biopsies to identify HPV types and HPV-16 was further evaluated by specific PCR real time.Among 87 cases, 12 (13.8%) had tumors harboring high risk HPV DNA and among 87 controls, 12 (13.8%) had high risk HPV DNA (OR:1.025 [CI:0.405:2.592]). Variables regarding consumption of alcohol and use of tobacco continued to characterize risk factors even after adjustments by presence or absence of high risk HPV.HPV was demonstrated to be frequently and similarly associated to normal and malignant esophageal tissues, but not as an independent risk factor to esophageal cancer.To contribute to the Brazilian population data on this subject, which is still contradictory.

Our goal was to describe prevalence of β-HPVs at three anatomic sites among 717 men from Brazil, Mexico and US enrolled in the HPV Infection in Men (HIM) Study. β-HPVs were genotyped using Luminex technology. Overall, 77.7%, 54.3% and 29.3% men were positive for any β-HPV at the genitals, anal canal, and oral cavity, respectively. Men from US and Brazil were significantly less likely to have β-HPV at the anal canal than men from Mexico. Older men were more likely to have β-HPV at the anal canal compared to younger men. Prevalence of β-HPV at the oral cavity was significantly associated with country of origin and age. Current smokers were significantly less likely to have β-HPV in the oral cavity than men who never smoked. Lack of associations between β-HPV and sexual behaviors may suggest other routes of contact such as autoinoculation which need to be explored further.

Background Human papillomavirus (HPV) prevalence in head and neck squamous cell carcinomas (HNSCC) diverges geographically. The reliability of using p16INK4a expression as a marker of viral infection is controversial in HNSCC. We evaluated HPV types and HPV-16 variants prevalence, and p16INK4a expression in HNSCC specimens provided by two different Institutions in São Paulo. Methods HPV DNA from formalin-fixed specimens was accessed by Inno-LiPA, HPV-16 variants by PCR-sequencing, and p16INK4a protein levels by immunohistochemistry. Results Overall, HPV DNA was detected among 19.4 % of the specimens (36/186). Viral prevalence was higher in the oral cavity (25.0 %, 23/92) then in other anatomical sites (oropharynx 14,3 %, larynx 13.7 %) when samples from both Institutions were analyzed together. HPV prevalence was also higher in the oral cavity when samples from both Institutions were analyzed separately. HPV-16 was the most prevalent type identified in 69.5 % of the HPV positive smaples and specimens were assigned into Asian-American (57.2 %) or European (42.8 %) phylogenetic branches. High expression of p16INK4a was more common among HPV positive tumors. Conclusion Our results support a role for HPV-16 in a subset of HNSCC.

OBJETIVO: Identificar os tipos de papilomavírus humano (HPV) nos portadores de carcinoma do canal anal (CCA), relacionando-os ao grau de diferenciação celular e estadiamento da lesão, em pacientes do Belém, Pará, entre 1998 e 2000. MÉTODOS: Foi realizado um estudo de caso-controle com 75 pacientes, divididos em: Grupo Teste, com 33 portadores de carcinoma do canal anal, e o Grupo Controle, com 42 portadores de doenças não-neoplásicas do canal anal. Os tipos virais foram identificados por PCR e dot blot. O teste exato de Fischer foi utilizado para avaliar a ocorrência de HPV. Adotou-se a tabela de contingência 3x2 para representar a distribuição dos tipos de HPV. Nos testes de hipóteses, foi prefixado o nível de significância α=0,05 para a rejeição da hipótese de nulidade. RESULTADOS: A prevalência do HPV foi significante entre os Grupos Teste (60,6%) e Controle (26,2%) (p=0,0027). Os tipos virais mais comuns foram 16 (42,4%) e 18 (15,2%). Observaram-se diferenças entre grupos na prevalência do HPV 16 (p=0,027) e 18 (p=0,043) no Grupo Teste, e o tipos 16 (19,0%,) e 18 em (2,4%) no Grupo Controle. No Grupo Teste, avaliou-se a distribuição dos tipos de HPV em relação ao estadiamento e ao grau de diferenciação celular, não apresentando diferenças estatisticamente significativas. CONCLUSÃO: O carcinoma de células escamosas do canal anal está associado à presença de HPV, e os tipos 16 e 18 são os mais frequentesOBJECTIVE: To identify the most predominant types of human papillomavirus (HPV) in carriers of the anal canal carcinoma (ACC), relating them to the cell differentiation and lesion staging degree, in patients from Belém, Pará, Brazil, between 1998 and 2000. METHODS: A case-control study was conducted with 75 patients, divided into Test Group, with 33 carriers of the anal canal carcinoma, and Control Group, with 42 carriers of non-neoplastic diseases of the anal canal. The methods employed to identify the viral types were the polymerase chain reaction and the dot blot. The Fisher's exact test was used to assess the HPV occurrence. The distribution of HPV types was analyzed by 3x2 contingency tables, representing the distribution of HPV types. For hypothesis testing, the significance level α=0.05 was previously established for rejection of the null hypothesis. RESULTS: HPV was significantly prevalent (p=0.0027) in the Test (60.6%) and Control Groups (26.2%) (p=0.0027). The most prevalent viral types were HPV 16 (42.4%) and 18 (15.2%). Significant differences related to the prevalence of HPV 16 and 18 were verified between both groups (p=0.027 and p=0.043, respectively). In the Control Group, HPV 16 was found in 19.0%, whereas HPV 18 was observed in 2.4%. In the Test Group, we evaluated the distribution of HPV types according to the staging and degree of cell differentiation, and found no significant differences between the results of the different groups. CONCLUSION: Squamous cell carcinoma of the anal canal is associated with the HPV presence, and the 16 and 18 types are the most frequently found

The serological detection of antibodies against human papillomavirus (HPV) antigens is a useful tool to determine exposure to genital HPV infection and in predicting the risk of infection persistence and associated lesions. Enzyme-linked immunosorbent assays (ELISAs) are commonly used for seroepidemiological studies of HPV infection but are not standardized. Intra- and interassay performance variation is difficult to control, especially in cohort studies that require the testing of specimens over extended periods. We propose the use of normalized absorbance ratios (NARs) as a standardization procedure to control for such variations and minimize measurement error. We compared NAR and ELISA optical density (OD) values for the strength of the correlation between serological results for paired visits 4 months apart and HPV-16 DNA positivity in cervical specimens from a cohort investigation of 2,048 women tested with an ELISA using HPV-16 virus-like particles. NARs were calculated by dividing the mean blank-subtracted (net) ODs by the equivalent values of a control serum pool included in the same plate in triplicate, using different dilutions. Stronger correlations were observed with NAR values than with net ODs at every dilution, with an overall reduction in nonexplained regression variability of 39%. Using logistic regression, the ranges of odds ratios of HPV-16 DNA positivity contrasting upper and lower quintiles at different dilutions and their averages were 4.73 to 5.47 for NARs and 2.78 to 3.28 for net ODs, with corresponding significant improvements in seroreactivity-risk trends across quintiles when NARs were used. The NAR standardization is a simple procedure to reduce measurement error in seroepidemiological studies of HPV infection.

The intratypic variability of HPVs 16 and 18 has been extensively studied and has been used as an important tool in epidemiological studies of viral transmission, persistence and progression to clinically relevant cervical lesions. Infections by non-European variants of HPVs 16 and 18 are associated with an increased risk for the development of high grade squamous intraepithelial lesions (HSIL). Our aim was to correlate the intratypic molecular variability of both HPV types and risk of persistent infection and lesion outcome in a cohort study conducted in Brazil. We characterized molecular variants of HPV types 16 and 18 by sequencing a fragment of the LCR, and of the E6 and L1 genes, for HPV-16 variants only. For both types, European variants composed the most prevalent and diverse group. Persistent infections with HPV-18 were associated with continuous detection of European variants. However, risk for simultaneous detection of HSIL and HPV DNA was higher in women harboring non-European variants of HPV-16. The same trend was observed with HSIL detected during follow-up. Our study confirms the association between non-European variants and risk of cervical neoplasia, and highlights the importance of their geographic distribution for cervical cancer risk assessment.

Background. We investigated the association between polymorphisms of human papillomavirus (HPV)-33 and squamous intraepithelial lesions (SILs). Methods. Endocervical specimens from 89 women infected with HPV-33, out of a total of 5347 recruited for 2 case-control and 2 cohort studies, were further analyzed by polymerase chain reaction sequencing of the long control region (LCR), E6, and E7. Results. Of the 89 samples, 64 were normal, 7 had low-grade SILs (including 3 determined by histopathologic analysis), 15 had high-grade SILs (HSILs, including 14 determined by histopathologic analysis), and 3 had an unknown diagnosis. Non-prototype-like LCR variants were significantly associated with HSILs (age- and study site-adjusted odds ratio [OR], 9.2 [95% confidence interval {CI}, 1.8-45.9]). The C7732G variation, which results in the loss of a putative binding site for the cellular upstream stimulatory factor, was associated with HSILs (age-and site-adjusted OR, 8.0 [95% CI, 1.5-42.8]). E6 and E7 polymorphisms were not associated with HSILs. Samples collected at 6-month intervals from 14 participants contained the same variant. The HPV-33 MT 1-0-0 variant carrying the G7584A variation was detected more frequently in women from Brazil (7/20 [35%]) than in women from Canada (1/65 [1.5%]; P = .001). Conclusion. Intratypic LCR variants of HPV-33 seem to vary geographically and to differ with respect to their oncogenic potential.

Background The genital skin of males hosts a diversity of HPV genotypes and uncharacterized HPV genotypes. Previously we demonstrated that a specific viral genotype was not identified in 14% of all genital specimens (i.e., HPV unclassified specimens) using the Roche Linear Array method. Our goal was to identify and assess the prevalence of individual HPV types among genital HPV unclassified specimens collected in the HIM Study population, at enrollment, and examine associations with socio-demographic and behavioral characteristics. Methods Genital skin specimens of men that were considered unclassified (HPV PCR positive, no genotype specified) at enrollment were typed by sequencing amplified PGMY09/11 products or cloning of PGMY/GP+ nested amplicons followed by sequencing. PGMY/GP+ negative specimens were further analyzed using FAP primers. HPV type classification was conducted through comparisons with sequences in the GenBank database. Results Readable nucleotide sequences were generated for the majority of previously unclassified specimens (66%), including both characterized (77%) and yet uncharacterized (23%) HPV types. Of the characterized HPV types, most (73%) were Beta [β]-HPVs, primarily from β-1 and β-2 species, followed by Alpha [α]-HPVs (20%). Smokers (current and former) were significantly more likely to have an α-HPV infection, compared with any other genus; no other factors were associated with specific HPV genera or specific β-HPV species. Conclusions Male genital skin harbor a large number of β-HPV types. Knowledge concerning the prevalence of the diverse HPV types in the men genital is important to better understand the transmission of these viruses. Electronic supplementary material The online version of this article (doi:10.1186/s12879-014-0677-y) contains supplementary material, which is available to authorized users.

Background Human papillomavirus type 6 (HPV-6) and HPV-11 are the etiological agents of approximately 90% of genital warts (GWs). The impact of HPV-6 genetic heterogeneity on persistence and progression to GWs remains undetermined. Methods HPV Infection in Men (HIM) Study participants who had HPV-6 genital swabs and/or GWs preceded by a viable normal genital swab were analyzed. Variants characterization was performed by polymerase chain reaction sequencing and samples classified within lineages (A, B) and sublineages (B1, B2, B3, B4, B5). Country- and age-specific analyses were conducted for individual variants; odds ratios and 95% confidence intervals for the risk of GWs according to HPV-6 variants were calculated. Results B3 variants were most prevalent. HPV-6 variants distribution differed between countries and case status. HPV-6 B1 variants prevalence was increased in GWs and genital swabs of cases compared to controls. There was difference in B1 and B3 variants detection in GW and the preceding genital swab. We observed significant association of HPV-6 B1 variants detection with GW development. Conclusions HPV-6 B1 variants are more prevalent in genital swabs that precede GW development, and confer an increased risk for GW. Further research is warranted to understand the possible involvement of B1 variants in the progression to clinically relevant lesions.

The HPV infection in men (HIM) study examines the natural history of genital HPV infection in men. Genotyping methods used in this study identify 37 α-HPV types; however, the viral type could not be identified in approximately 22% of male genital specimens that were HPV PCR positive. Our aim was to genotype HPV-unclassified specimens by sequencing PGMY09/11, GP5+/6+ or FAP59/64 PCR products. Using this approach we were able to detect 86 unique HPV types among 508 of 931 specimens analyzed. We report for the first time the presence of a broad range of α-, β- and γ-HPV at the male genitals.

Purpose: To investigate the presence of human papillomavirus (HPV) DNA in tumour tissue from patients with unilateral retinoblastoma. Methods: Samples of paraffin-embedded tumour tissue from 43 children with unilateral retinoblastoma were collected to investigate the presence of HPV DNA using polymerase chain reaction (PCR) and dot blot hybridization. Results: Oncogenic HPV DNA types 16 and 35 were detected in 12 (27.9%) of 43 tumour specimens. A higher frequency of differentiated tumours (63.3%) was observed among the HPV-positive tumours. Conclusions: Future studies are necessary to demonstrate an association between HPV and sporadic retinoblastoma.

OBJETIVO: Identificar os tipos de papilomavírus humano (HPV) nos portadores de carcinoma do canal anal (CCA), relacionando-os ao grau de diferenciação celular e estadiamento da lesão, em pacientes do Belém, Pará, entre 1998 e 2000. MÉTODOS: Foi realizado um estudo de caso-controle com 75 pacientes, divididos em: Grupo Teste, com 33 portadores de carcinoma do canal anal, e o Grupo Controle, com 42 portadores de doenças não-neoplásicas do canal anal. Os tipos virais foram identificados por PCR e dot blot. O teste exato de Fischer foi utilizado para avaliar a ocorrência de HPV. Adotou-se a tabela de contingência 3x2 para representar a distribuição dos tipos de HPV. Nos testes de hipóteses, foi prefixado o nível de significância α=0,05 para a rejeição da hipótese de nulidade. RESULTADOS: A prevalência do HPV foi significante entre os Grupos Teste (60,6%) e Controle (26,2%) (p=0,0027). Os tipos virais mais comuns foram 16 (42,4%) e 18 (15,2%). Observaram-se diferenças entre grupos na prevalência do HPV 16 (p=0,027) e 18 (p=0,043) no Grupo Teste, e o tipos 16 (19,0%,) e 18 em (2,4%) no Grupo Controle. No Grupo Teste, avaliou-se a distribuição dos tipos de HPV em relação ao estadiamento e ao grau de diferenciação celular, não apresentando diferenças estatisticamente significativas. CONCLUSÃO: O carcinoma de células escamosas do canal anal está associado à presença de HPV, e os tipos 16 e 18 são os mais frequentes OBJECTIVE: To identify the most predominant types of human papillomavirus (HPV) in carriers of the anal canal carcinoma (ACC), relating them to the cell differentiation and lesion staging degree, in patients from Belém, Pará, Brazil, between 1998 and 2000. METHODS: A case-control study was conducted with 75 patients, divided into Test Group, with 33 carriers of the anal canal carcinoma, and Control Group, with 42 carriers of non-neoplastic diseases of the anal canal. The methods employed to identify the viral types were the polymerase chain reaction and the dot blot. The Fisher's exact test was used to assess the HPV occurrence. The distribution of HPV types was analyzed by 3x2 contingency tables, representing the distribution of HPV types. For hypothesis testing, the significance level α=0.05 was previously established for rejection of the null hypothesis. RESULTS: HPV was significantly prevalent (p=0.0027) in the Test (60.6%) and Control Groups (26.2%) (p=0.0027). The most prevalent viral types were HPV 16 (42.4%) and 18 (15.2%). Significant differences related to the prevalence of HPV 16 and 18 were verified between both groups (p=0.027 and p=0.043, respectively). In the Control Group, HPV 16 was found in 19.0%, whereas HPV 18 was observed in 2.4%. In the Test Group, we evaluated the distribution of HPV types according to the staging and degree of cell differentiation, and found no significant differences between the results of the different groups. CONCLUSION: Squamous cell carcinoma of the anal canal is associated with the HPV presence, and the 16 and 18 types are the most frequently found

Human papillomavirus (HPV) genotypes cannot fully explain the histological diagnosis of women with glandular abnormalities detected by cervical smear. Thus, this study was designed to analyze the distribution of HPV-16 and HPV-18 variants in women referred because of atypical glandular cells and adenocarcinoma in situ in their cervical smears and its association with histological results. Twenty-four women with HPV-16 and 6 with HPV-18, selected from 160 women with cervical smears suggestive of glandular abnormalities, were included. Histological results showed cervicitis (1 case), squamous neoplasia (18 cases), glandular neoplasia (7 cases), and glandular neoplasia associated with a squamous component (4 cases). Among the 24 cases presenting HPV-16, the European variant was detected in 15 (62%) and the Asian American in 9 (38%). Among the 15 cases associated with the European variant, 14 (93%) presented squamous neoplasia and 1 (7%) invasive adenocarcinoma. Asian-American HPV-16 variants were significantly associated with histological diagnosis of glandular neoplasia alone (odds ratio, 9.3 [1.4-60.2]) or associated with squamous neoplasia (odds ratio, 18.7 [1.5-232.3]). Adenocarcinomas were detected in 4 of 6 HPV-18-positive cases, being 2 cases had the European variant, 1 had the Asian Amerindian variant, and 1 had the African variant. The association of HPV-16 with squamous or glandular neoplasia is explained by its variants. In this study, squamous neoplasia was related to the European variant of HPV-16, whereas glandular neoplasia was related to the Asian-American variant. Glandular neoplasia is associated with HPV-18, but the results of our analysis of its variants were inconclusive.

ContextHuman papillomavirus (HPV) infection is believed to be the central cause of cervical cancer, although most of the epidemiological evidence has come from retrospective, case-control studies, which do not provide information on the dynamics of cumulative or persistent exposure to HPV infection.ObjectiveTo assess the risks of cervical neoplasia related to prior persistent HPV infections.Design and SettingLongitudinal study of the natural history of HPV infection and cervical neoplasia in women residing in the city of Sao Paulo, Brazil, which was conducted between November 1993 and March 1997 and involved repeated measurements of HPV and lesions with follow-up until June 2000.ParticipantsA total of 1611 women with no cytological lesions at enrollment and HPV test results available from the first 2 visits.Main Outcome MeasureCervical specimens taken for Papanicolaou cytology and HPV testing every 4 months in the first year and twice yearly thereafter. Incident cervical cancer precursor lesions ascertained by expert review of all cytology smears.ResultsThe incidence rate of squamous intraepithelial lesions (SILs) was 0.73 per 1000 women-months (95% confidence interval [CI], 0.5-0.9) among women free of HPV at the 2 initial visits and 8.68 (95% CI, 2.3-15.1) among women with HPV type 16 or 18 infections persisting over both visits. Relative to those negative for HPV oncogenic types at both initial visits, the relative risk (RR) of incident SIL was 10.19 (95% CI, 5.9-17.6) for persistent infections with any known oncogenic HPV types. The equivalent RR of incident high-grade SIL was 11.67 (95% CI, 4.1-33.3). The RRs of lesions were considerably higher for persistent infections with HPV type 16 or 18.ConclusionA strong relationship exists between persistent HPV infections and SIL incidence, particularly for HPV types 16 and 18.