Methylation at 3'LCR of HPV16 can be affected by patient age and disruption of E1 or E2 genes.

CpG methylation at early promoter of HPV16 DNA, in the 3' end of the Long Control Region (3'LCR), has been associated to the presence of episomal forms of viral genome and, consequently, intact E1 and E2 ORFs. The DNA methylation would block the access of E2 viral protein to the E2 binding sites at early-promoter. However, is still unclear if methylation at 3'LCR of HPV16 DNA can also vary depending of other tumor characteristics in addition to viral DNA physical state. In this study, we evaluate whether the methylation level at the five CpG located at 3'LCR of HPV16 is associated to patient age and E1 and/or E2 ORFs integrity. DNA pyrosequencing was used to measure the methylation level in 69 invasive cervical cancer samples obtained from biopsies of patients attended at Brazilian National Institute of Cancer (INCA). PCR amplifications were performed to assess disruption status of E1 and E2 genes of HPV16. The methylation average per sample ranged widely, from <1 to 88.00%. Presence of intact E1/E2 genes and patient age were positively associated with average methylation in both bivariate analyses (p=0.003 and p=0.006, respectively), and multivariate analysis (p=0.002 and p=0.021, respectively), adjusted for tumor type (squamous cell carcinomas or adenocarcinomas) and HPV16 lineage. These findings showed that presence of intact E1/E2 open reading frames was associated with high levels of DNA methylation, and older patients showed higher levels of methylation than younger ones independently of viral genome disruption.

High Risk Human Papillomavirus Infection of the Foreskin in Asymptomatic Men and Patients with Phimosis.

PURPOSE: There has been increasing interest in understanding the natural history of HPV and the diseases that it causes in men. HPV infection is strongly associated with penile cancer, lack of neonatal circumcision and phimosis. We investigated the incidence of HPV infection in asymptomatic men and patients with phimosis. MATERIALS AND METHODS: We assessed 110 asymptomatic men and 30 patients who underwent circumcision due to phimosis. DNA was extracted from swabbed samples collected from asymptomatic men and from foreskin samples collected at circumcision. Polymerase chain reaction using consensus primers for detecting HPV-MY09/11 was performed to detect generic HPV DNA. HPV genotyping was done by polymerase chain reaction amplification with primers for the E6 gene DNA sequences HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV35, HPV45 and HPV58. RESULTS: HPV was present in 46.66% of patients with phimosis, of whom 50% had high risk HPV genotypes. Of asymptomatic cases 16.36% were HPV positive but only 1 sample showed high risk HPV. We detected a significantly high rate of HPV genital infection in patients presenting with phimosis compared with asymptomatic men (p = 0.00167). The prevalence of high risk HPV genotypes in patients with phimosis was also statistically significant (p = 0.0004). CONCLUSIONS: We found a robust association between phimosis and the genital HPV prevalence in men and a significant frequency of high risk HPV. Other studies are needed to investigate the occurrence of factors that can increase the incidence of penile carcinoma and determine its impact on female genital infection in cervical cancer.

Prevalence of human papillomavirus infection in the genital tract determined by hybrid capture assay

Human Papillomavirus (HPV) infection is the most prevalent sexually-transmitted virus worldwide. It is known to be the etiological agent of cervical cancer and cervical intraepithelial neoplasia (CIN). Consequently, there is strong motivation to evaluate HPV testing in cervical cancer screening. Recently developed, the second generation of the hybrid capture test (HCA II) is a non-radioactive, relatively rapid, hybridization assay, designed to detect 18 HPV types divided into high and low-risk groups. We evaluated 7,314 patients (5,833 women and 1,481 men) for HPV infection by HCA II. Among them, 3,008 (41.1 percent) presented HPV infection: 430 (14.2 percent) had HPV DNA of low risk for cancer, 1,631 (54.2 percent) had high risk HPV types and 947 (31.5 percent) had both types. The prevalence in females was 44.9 percent. The prevalence of HPV DNA in the group for which cytological results were available was slightly higher: 55.3 percent (1007/1824). Significant differences were detected in the frequency of HPV infection of the cervix between normal cases and those with high-grade squamous-intraepithelial lesions (HSIL)(P<0.0001). Among males, the prevalence was 26.2 percent, composed of 9.1 percent in Group A, 9.7 percent in Group B and 7.4 percent with multiple infections. We observed that male prevalence was lower and that low-risk types were more frequent than in females. HPV viral load was significantly greater in SILs than in normal or inflammatory cases (P<0.0001), suggesting an association between high viral load values and risk of SIL. Because of high costs, the HCA II test cannot be recommended for routine mass screening for cervical infection in poor countries. Nevertheless, it was found to be a useful tool, when combined with cytology, discovering high-risk infections in apparently normal tissues and revealing silent infections that may be responsible for the maintenance of HPV in the general population. These findings point...

Infecções causadas pelos papilomavírus humanos: atualização sobre aspectos virológicos, epidemiológicos e diagnóstico / Human papillomavirus infection: update on virological and epidemiological aspects and diagnosis

Papilomavírus humanos são pequenos vírus de DNA capazes de infectar epitélio da pele e mucosas. Mais de 100 tipos já foram descritos, sendo que cerca de 40 causam doenças sexualmente transmissíveis. É também no trato genital que alguns tipos exercem poder oncogênico, sendo 13 considerados agentes etiológicos do câncer cervical, responsável pela morte de 200.000 mulheres em todo o mundo, anualmente. O diagnóstico disponível para o controle das DST e câncer causados pelos HPV baseia-se no tripé colposcopia, citologia e histopatologia, podendo ser confirmado laboratorialmente por técnicas de biologia molecular. Dentre elas: PCR e captura do híbrido. Recentemente, duas vacinas profiláticas contra os tipos 6, 11, 16 e 18 (Merck) e 16 e 18 (Glaxo) foram desenvolvidas. São compostas de VLPs (Vírus-like particles) expressas em leveduras. Resultados promissores vêm sendo publicados, embora alguns mecanismos imunológicos de defesa contra o HPV não estejam ainda esclarecidos.

Human papillomavirus are small DNA viruses capable of infecting the epithelium of skin and mucosa. More than a hundred types have already been described, and nearly 40 promote sexually transmitted diseases. It is in the genital tract that 13 HPV types exert their oncogenic potential, being responsible by the death of 200,000 women worldwide, annualy. The available diagnosis to STD and cancer control is based upon the colposcopy, cytology and histopathology tools, and can be laboriatorially confirmed by molecular biology techniques: PCR and hybrid capture. Recently, two prophylactic vaccines against HPV types 6, 11, 16 e 18 (Merck) and 16 e 18 (Glaxo) were developed. They are composed by VLP (Virus-like particles) expressed in yeasts. Promissing results have been published. Nevertheless, immunological mechanisms for human defense are not well elucidated.

Prevalence of human papillomavirus infection in the genital tract determined by hybrid capture assay.

Human Papillomavirus (HPV) infection is the most prevalent sexually-transmitted virus worldwide. It is known to be the etiological agent of cervical cancer and cervical intraepithelial neoplasia (CIN). Consequently, there is strong motivation to evaluate HPV testing in cervical cancer screening. Recently developed, the second generation of the hybrid capture test (HCA II) is a non-radioactive, relatively rapid, hybridization assay, designed to detect 18 HPV types divided into high and low-risk groups. We evaluated 7,314 patients (5,833 women and 1,481 men) for HPV infection by HCA II. Among them, 3,008 (41.1%) presented HPV infection: 430 (14.2%) had HPV DNA of low risk for cancer, 1,631 (54.2%) had high risk HPV types and 947 (31.5%) had both types. The prevalence in females was 44.9%. The prevalence of HPV DNA in the group for which cytological results were available was slightly higher: 55.3% (1007/1824). Significant differences were detected in the frequency of HPV infection of the cervix between normal cases and those with high-grade squamous-intraepithelial lesions (HSIL)(P<0.0001). Among males, the prevalence was 26.2%, composed of 9.1% in Group A, 9.7% in Group B and 7.4% with multiple infections. We observed that male prevalence was lower and that low-risk types were more frequent than in females. HPV viral load was significantly greater in SILs than in normal or inflammatory cases (P<0.0001), suggesting an association between high viral load values and risk of SIL. Because of high costs, the HCA II test cannot be recommended for routine mass screening for cervical infection in poor countries. Nevertheless, it was found to be a useful tool, when combined with cytology, discovering high-risk infections in apparently normal tissues and revealing silent infections that may be responsible for the maintenance of HPV in the general population. These findings point to the need for close and careful management of patients, thereby reducing overtreatment, allowing analysis of both sexual partners and finally contributing to the control of genital infections associated with a risk for cancer.

Detecção de papilomavírus humanos pela técnica de captura do híbrido no trato genital masculino / Human papillomavirus detection by the hybrid capture assay in the male genital tract

Estudo das infecções genitais causadas pelo HPV, determinando a prevalência dos tipos de HPV noa pacientes masculinos

Comparative analysis of the polymerase chain reaction and the hybrid capture assay for the detection of human papillomavirus infection

Avalia o emprego de métodos moleculares a fim de comprovar a presença dos HPV no trato genital para utilização em combinação com a citopatologia, método utilizado para rastreamento das lesões causadas pelo HPV

Estudo de infecçöes por papilomavírus humanos em pacientes do sexo feminino, detectados pela técnica de captura do híbrido: levantamento dos casos / Study of human papilomavirus infections in feminine sex patients, detected by hybrid capture assay in: survey of the cases

Estudo das infecçöes genitais causadas pelo HPV, determinando a prevalência dos tipos de HPV em pacientes do sexo feminino, atendidas pelo Laboratório Sérgio Franco do Estado do Rio de Janeiro, no ano de 2001, pela técnica de captura do híbrido