Characterisation of complete high- and low-risk human papillomavirus genomes isolated from cervical specimens in southern Brazil

The classification of human papillomavirus (HPV) intratypic lineages by complete genome sequencing is a determinant in understanding biological differences in association with this disease. In this work, we have characterised complete HPV genomes from southern Brazil. Fifteen cervicovaginal Pap smear negative samples previously categorised as HPV-positive were sequenced using ultradeep sequencing, and 18 complete genomes from 13 different HPV types were assembled. Phylogenetic and genetic distance analyses were performed to classify the HPV genomes into lineages and sublineages. This is the first report describing the distribution of HPV intratype lineages of high and low oncogenic risk in asymptomatic women from southern Brazil.

Papillomavirus infections in the oral and genital mucosa of asymptomatic women

Abstract Human papillomavirus (HPV) has been found in several regions of the body, including the oral cavity. Recently, this virus has been associated with oropharyngeal cancer, but little is known about HPV transmission to the oral cavity. We carried out a study to investigate concurrent oral and cervical infections in 76 asymptomatic women attending a healthcare program. Demographic and behavior data were obtained through a structured questionnaire. Oral and cervical mucosa scrapings were collected and stored for DNA extraction. HPV DNA amplification was performed by polymerase chain reaction assay (PCR) using both primers My09/My11 and FAP59/64, followed by HPV typing with restriction fragment length polymorphism analysis (RFLP) and sequencing. The data collected revealed no risk factors for HPV infection in these 76 women. HPV prevalence of 9.2 and 5.3% was found in cervical and oral mucosa, respectively. Concurrent infections by discordant types were detected in one case only. Sequencing procedures allowed us to detect a new putative HPV 17 subtype from the Betapapillomavirus genus. Our results support the view that cervical and oral HPV infections are independent events. The observed low prevalence of both oral and cervical HPV infections could be associated with attendance in a healthcare program.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

The impact of glucocorticoids and anti-cd20 therapy on cervical human papillomavirus infection risk in women with systemic lupus erythematosus

OBJECTIVE: To identify the prevalence and factors associated with cervical human papillomavirus infection in women with systemic lupus erythematosus METHODS: This cross-sectional study collected traditional and systemic lupus erythematosus-related disease risk factors, including conventional and biologic therapies. A gynecological evaluation and cervical cytology screen were performed. Human papillomavirus detection and genotyping were undertaken by PCR and linear array assay. RESULTS: A total of 148 patients were included, with a mean age and disease duration of 42.5±11.8 years and 9.7±5.3 years, respectively. The prevalence of squamous intraepithelial lesions was 6.8%. The prevalence of human papillomavirus infection was 29%, with human papillomavirus subtype 59 being the most frequent. Patients with human papillomavirus were younger than those without the infection (38.2±11.2 vs. 44.2±11.5 years, respectively; p = 0.05), and patients with the virus had higher daily prednisone doses (12.8±6.8 vs. 9.7±6.7 mg, respectively; p = 0.01) and cumulative glucocorticoid doses (14.2±9.8 vs. 9.7±7.3 g, respectively; p = 0.005) compared with patients without. Patients with human papillomavirus infection more frequently received rituximab than those without (20.9% vs. 8.5%, respectively; p = 0.03). In the multivariate analysis, only the cumulative glucocorticoid dose was associated with human papillomavirus infection. CONCLUSIONS: The cumulative glucocorticoid dose may increase the risk of human papillomavirus infection. Although rituximab administration was more frequent in patients with human papillomavirus infection, no association was found. Screening for human papillomavirus infection is recommended in women with systemic lupus erythematosus. .

Evidence of a higher prevalence of HPV infection in HTLV-1-infected women: a cross-sectional study / Evidência de maior prevalência de infecção pelo HPV em mulheres infectadas pelo HTLV-1: um estudo de corte transversal

INTRODUCTION:HTLV-1 infection increases susceptibility to other infections. Few studies have addressed the co-infection between HPV and HTLV-1 and the immune response involved in this interaction. The aim of this study was to determine the prevalence of cervical HPV infection in HTLV-1-infected women and to establish the risk factors involved in this co-infection. METHODS: A cross-sectional study was carried out in Salvador, Brazil, between September 2005 and December 2008, involving 50 HTLV-1-infected women from the HTLV Reference Center and 40 uninfected patients from gynecological clinic, both at the Bahiana School of Medicine. HPV infection was assessed using hybrid capture. HTLV-1 proviral load was quantified using real-time polymerase chain reaction (PCR). RESULTS: The mean age of HTLV-1-infected women (38 ± 10 years) was similar to that of the control group (36 ± 13 years). The prevalence of HPV infection was 44% in the HTLV-1-infected group and 22.5% in uninfected women (p = 0.03). HTLV-1-infected women had lower mean age at onset of sexual life (17 ± 3 years versus 19 ± 3 years; p = 0.03) and greater number of lifetime partners compared with the control group (4 ± 3 versus 2 ± 1; p < 0.01). In the group of HTLV-1-infected patients, there was neither difference in HTLV-1 proviral load between HPV-infected women and the uninfected. CONCLUSIONS: The prevalence of HPV infection was higher in HTLV-1-infected women. Further studies should be performed to evaluate the progression of this co-infection.

INTRODUÇÃO:A infecção pelo HTLV-1 aumenta a susceptibilidade para outras infecções. Poucos estudos avaliaram a co-infecção entre HPV/HTLV-1 e a resposta imune envolvida nesta interação. O objetivo deste trabalho é determinar a prevalência de infecção cervical pelo HPV em mulheres infectadas pelo HTLV-1 e estabelecer os fatores de risco envolvidos nesta co-infecção. MÉTODOS: Um estudo de corte transversal foi conduzido em Salvador, Brasil, entre setembro de 2005 e dezembro de 2008, envolvendo 50 mulheres infectadas pelo HTLV-1, acompanhadas no Centro de Referência de HTLV e 40 mulheres não infectadas, acompanhadas no Serviço de Ginecologia, ambos na Escola Bahiana de Medicina. A infecção pelo HPV foi confirmada pela Captura Híbrida. A carga proviral do HTLV-1 foi quantificada pelo PCR em tempo real. RESULTADOS: A média de idade das mulheres infectadas pelo HTLV-1 (38±10 anos) foi semelhante ao do grupo controle (36±13 anos). A prevalência de infecção pelo HPV foi 44% nas mulheres infectadas pelo HTLV-1 e de 22,5% no grupo controle (p=0,03). Mulheres infectadas pelo HTLV-1 informaram menor idade de início de vida sexual (17±3 anos versus 19±3 anos; p=0,03) e maior número de parceiros sexuais, em relação ao grupo controle (4±3 versus 2±1; p<0,01). No grupo de mulheres infectadas pelo HTLV-1, não se observou diferença entre a carga proviral do HTLV-1 entre as mulheres infectadas pelo HPV e as não infectadas. CONCLUSÕES: A prevalência de infecção pelo HPV foi maior em mulheres infectadas pelo HTLV-1. Novos estudos devem ser realizados para avaliar a progressão desta co-infecção.

Infecção oral pelo HPV em mulheres com lesão escamosa de colo uterino no sistema prisional da cidade de São Paulo, Brasil / Oral infection by the Human Papilloma Virus in women with cervical lesions at a prison in São Paulo, Brazil

O carcinoma de cabeça e pescoço é 6ª maior causa de mortes por neoplasia no mundo. Nas últimas décadas, tem-se associado a relação da infecção pelo Papilomavírus Humano (HPV) e seu envolvimento na etiologia desta doença, bem como acontece com o câncer de colo de útero. OBJETIVO: A caracterização molecular dos tipos de HPV diagnosticados na mucosa oral de mulheres que apresentavam alterações citológicas compatíveis com lesão escamosa no colo uterino. MÉTODOS: Foram estudadas 409 amostras cérvico-vaginais e de cavidade oral de mulheres internas no Presídio Feminino da cidade de São Paulo. A correlação entres lesões cervicais e orais foram avaliadas em 27 mulheres que apresentavam lesões pré-malignas e malignas no colo uterino pela caracterização molecular dos tipos de HPV por PCR/ RFLP e Sequenciamento. RESULTADOS: Das 27 (6,67%) amostras compatíveis com LSIL e HSIL no colo uterino, 22 (81,48%) apresentaram infecção pelo HPV de alto risco oncogênico, sendo o HPV 59 o mais prevalente, dentre elas, três amostras (11,1%) evidenciaram alterações celulares compatíveis com displasia leve na cavidade oral. CONCLUSÃO: Nosso estudo sugere uma relação entre o desenvolvimento de lesões da cavidade oral e a infecção pelo HPV, independentemente do tipo viral presente.

Carcinoma of the head and neck is the 6th cause of death by cancer in the world. In recent decades the human papillomavirus (HPV) has been implicated in the etiology of this disease. OBJECTIVE: To characterize the types of HPV detected in the oral mucosa in women with cytological abnormalities suggesting intraepithelial squamous lesions in the uterine cervix. METHODS: four-hundred-nine cervical-vaginal and oral pap-smears of women interned in a Female Prison in São Paulo were examined. The relationship between cervical and oral lesion was analyzed by PCR/RFLP and DNA sequencing. RESULTS: Of 27 (6.67%) specimens showing cervical cytological abnormalities suggesting LSIL and HSIL, 22 (81.48%) had oncogenic high-risk HPV infection, of which HPV 59 was the most prevalent. Three (11.1%) samples showed cytological changes suggesting mild dysplasia in the oral cavity. CONCLUSION: Our study suggests an association between carcinoma of the oral cavity and HPV infection, regardless of the virus type.

Factores de riesgo de alteraciones citológicas del cuello uterino en mujeres chilenas: Un estudio de casos y controles / Risk factors associated with abnormal cervical cytology among Chilean women: A case control study

Background: Cervical cancer is the third cause of cancer death among Chilean women, affecting mainly women from low socioeconomic status. Aim: To determine main risk factors (RF) including human papilomavirus (HPV) types associated with abnormal cervical cytology (Atypical Squamous Cells of Undetermined Significance or ASCUS) among Chilean women from low socioeconomic status in Santiago, Chile. Material and Methods: A random population based sample of616 women from La Pintana (a low-income district in Santiago) participated in 2001 in a HPV prevalence study and were re-evaluated in 2006 through a risk factors questionnaire, Papanicolaou test and DNA detection for HPV. The Papanicolaou test was analyzed in Santiago and HPV analysis (PCR_GP5+/GP6+) was conducted in Vrije University, Amsterdam. Cases included 42 women with cervical lesions and controls included 574 women with normal cytology during the period 2001-2006. Logistic regression with uni and multivariate analysis was performed to identify RF for cervical lesions. Results: During the study period, there was a significant increase in the proportion of single women, from 8.3 to 14.8 percent (p < 0.05), of women with 3 or more sexual partners from 8.9 to 13.3 and of women high risk HPV, from 9.1 to 14.3 percent. The proportion of abnormal Papanicolaou tests remained stable (3.08 and 3.9 percent > ASCUS). High risk HPV was the most significant factor associated with cervical lesions (odds ratio (OR) = 9.695 percent> confidence intervals (CI) = 4.4-21.1) followed by oral contraceptive use (OR = 2.58 95 percent> CI= 1.2-5.7). Among women infected by high risk HPV, the use of oral contraceptives was a risk factor while compliance with screening was protective for cervical lesions. Conclusions: From 2001 to 2006, there was an increase in the proportion of women with high-risk HPV infections.

The environmental cofactors in carcinogenesis in high risk HPV/HIV-positive women

The objective of the present study was to assess the presence of human papilloma virus (HPV) in HIV-infected women, with comparison between the Papanicolaou cytologic technique and the molecular PCR technique, as well as to determine the type of HPV, to measure cellular immunocompetence and to identify the presence of risk factors for the acquisition of HPV infection. Thirty HIV-infected women were selected. Vaginal and endocervical samples were collected from 27 of them. The smears were examined by 3 experienced cytologists to diagnose the presence of HPV by the Papanicolaou technique and the results were compared to HPV detection and typing by PCR. HPV-infected patients were interviewed in order to identify the presence of risk factors for the acquisition of the virus. Eight of the 27 patients analyzed (29 percent) presented HPV in endocervical samples submitted to PCR, 6 of them (75 percent) presented HPV involving a high risk of development of cervical cancer. For 5 of these patients, the cytologic diagnosis was not confirmed by PCR. When cellular immunocompetence was related to HPV infection, PCR revealed a diagnosis of HPV in 37.50 percent of the patients at intermediate risk for HPV infection and in 83.33 percent of the patients at high risk for HPV infection. These immunologically compromised HPV-infected patients are at higher risk of developing cervical neoplasia. We showed here that PCR is adequate for HPV detection and that, if only the Papanicolaou method is used for the follow-up of these patients, we will not provide good prevention of cervical cancer.

Human papillomavirus status and cervical abnormalities in women from public and private health care in Rio de Janeiro State, Brazil / Papilomavírus humano e anormalidades cervicais em mulheres do sistema de saúde privado e público no Estado do Rio de Janeiro, Brasil

This article reports the HPV status and cervical cytological abnormalities in patients attended at public and private gynecological services from Rio de Janeiro State. It also comments the performance of each HPV DNA tests used. A set of 454 women from private health clinics was tested by routine Capture Hybrid II HPV DNA assay. Among these, 58.4 percent presented HPV and nearly 90 percent of them were infected by high risk HPV types. However, this group presented few premalignant cervical lesions and no invasive cervical cancer was registered. We also studied 220 women from low income class attended at public health system. They were HPV tested by polymerase chain reaction using My09/11 primers followed by HPV typing with E6 specific primers. The overall HPV prevalence was 77.3 percent. They also showed a high percentage of high squamous intraepithelial lesion-HSIL (26.3 percent), and invasive cervical carcinoma (16.3 percent). HPV infection was found in 93.1 percent and 94.4 percent of them, respectively. The mean ages in both groups were 31.5 and 38 years, respectively. In series 1, HPV prevalence declined with age, data consistent with viral transient infection. In series 2, HPV prevalence did not decline, independent of age interval, supporting not only the idea of viral persistence into this group, but also regional epidemiological variations in the same geographic area. Significant cytological differences were seen between both groups. Normal and benign cases were the most prevalent cytological findings in series 1 while pre-malignant lesions were the most common diagnosis in the series 2. HPV prevalence in normal cases were statistically higher than those from series 1 (p < 0.001), indicating a higher exposure to HPV infection. Women from both samples were referred for previous abnormal cytology. However, socio-demographic evidence shows that women from series 1 have access to treatment more easily and faster than women from series 2 before the development of pre-malignant lesions...

Este artigo analisa a infecção por HPV e anormalidades citológicas cervicais encontradas em pacientes atendidas em serviços ginecológicos dos sistemas de saúde público e privado do estado do Rio de Janeiro. O trabalho também avalia os testes utilizados para detecção de DNA do HPV em cada população estudada. Um grupo de 454 mulheres oriundas de serviços da rede privada de saúde foi testado por Captura do Híbrido II. Destas, 58,4 por cento apresentaram infecção por HPV e cerca de 90 por cento delas estavam infectadas por HPV de alto risco. Este grupo, entretanto, apresentava poucos casos de lesões cervicais pré-malígnas e nenhum caso de câncer. Estudamos, também, 220 mulheres de baixo nível econômico atendidas no serviço de saúde pública que foram testadas para HPV pela reação da polimerase em cadeia utilizando-se os oligonucleotídeos My09/My11. A identificação dos tipos foi efetuada por amplificação com oligonucleotídeos específicos para a região E6 do genoma viral. A prevalência de HPV nesta população foi de 77.3 por cento, observando-se uma alta porcentagem de casos de neoplasias intraepiteliais cervicais de alto grau (26,3 por cento) e de carcinoma cervical invasivo (16,3 por cento). A infecção por HPV foi achada em, respectivamente, 93,1 por cento e 94,4 por cento destes casos. A média de idade em ambos os grupos era de 31,5 e 38 anos, respectivamente. Na série 1, a prevalência da infecção por HPV decresce com a idade, enquanto na série 2 ela não desaparece, dando suporte não só à idéia de persistência viral neste grupo, mas também a variações epidemiológicas na mesma área geográfica. Diferenças significativas foram vistas nos dois grupos. Casos normais e benignos foram incidentes na série 1, enquanto as lesões malígnas predominaram na série 2. Ao contrário, casos normais infectados por HPV eram prevalentes na série 2 (p < 0.001), indicando maior exposição ao vírus. Embora as mulheres de ambos os grupos tenham sido incluídas no estudo por apresentarem citologia anormal, evidências sócio-demográficas demonstram que mulheres da série 1 tem acesso mais fácil e rápido ao tratamento do que as mulheres da série 2 antes que as lesões pré-malígnas se desenvolvam...

Analysis of molecular biology techniques for the diagnosis of human papillomavirus infection and cervical cancer prevention

The objective of the present study was to evaluate the usefulness of molecular methodologies to access human papillomavirus genome in the genital tract. Samples from 136 women aged 17 to 52 years old obtained from the Dr. Sérgio Franco Laboratories between 2000 and 2001, were analyzed by the hybrid capture assay and amplified by PCR with generic primers MY09/MY11 and specific primers for types 16, 18, 31, 33, 35, 58. Viral genome was detected in 71.3 percent of the samples by hybrid capture and 75 percent by amplification. When cytopathology was used as a reference method for screening lesions, hybrid capture (p=0) and amplification (p=0.002) presented positive association. The 3 methods showed absolute agreement when cytopathology confirmed papillomavirus infection and high grade intraepithelial lesion. Disagreements occurred for 10 cases: seven inflammatory cases positive by PCR and negative for hybrid capture and 3 low squamous intraepithelial lesions positive for hybrid capture but negative for amplification. In conclusion, hybrid capture was shown to be sensitive and specific enough for use in clinical routines. Moreover, the evaluation of viral load values obtained by this method were shown to be related to the severity of the lesion and merit further studies to analyze the possible association with risk of progression to malignancy.

O objetivo do nosso trabalho foi avaliar o emprego de métodos moleculares para comprovar a presença dos papilomavírus humanos no trato genital. Amostras de 136 pacientes com idades entre 17 e 52 anos, coletadas nos Laboratórios Dr. Sérgio Franco entre 2000 e 2001, foram analisadas pelas técnicas de captura híbrida e amplificação pela reação em cadeia da polimerase com primers genéricos MY09/MY11 e específicos para os tipos 16, 18, 31, 33, 35, 58. O genoma viral foi detectado em 71,3 por cento dessas amostras pela captura híbrida e 75 por cento pela reação em cadeia da polimerase. Quando a citopatologia foi usada como método de referência para rastreamento das lesões, a captura (p=0) e a amplificação (p=0,002) demonstraram associação positiva. Os três testes demonstraram concordância absoluta quando a citopatologia diagnosticou processo compatível com papilomavírus ou lesão intraepitelial de alto grau. Dez casos discordantes ocorreram: sete casos de citologia inflamatório positivos na reação em cadeia da polimerase mas negativos na captura híbrida e 3 casos de lesão de baixo grau positivas na captura híbrida e negativas pela reação em cadeia da polimerase. Concluímos que a captura híbrida apresentou sensibilidade e especificidade adequadas para uso clínico. Avaliação da carga viral obtida por esta metodologia relacionou-se com a severidade da lesão e merece estudos adicionais a fim de determinar seu valor prognóstico para o câncer.

Características epidemiológicas en mujeres con esterilidad y lesiones cervicales con y sin infección por virus de papiloma humano (VPH). Estudio comparativo / Epidemiologic traits in women with sterility and cervical lesions with or without infection by human papilloma virus (VPH). Comparative study

Las infecciones por el Virus del Papiloma Humano (VPH) son una de las entidades que con mayor frecuencia causan lesiones cervicales de morfología variada. Su influencia sobre el fenómeno reproductivo humano no es completamente clara. Previamente, hemos informado en un grupo de mujeres con esterilidad asociada a lesiones cervicales por VPH que la tasa de embarazo espontáneo posterior al tratamiento de la infecciín fue de 53.5 por ciento. En este informe se destacó la necesidad de hacer un estudio controlado. Comparar las principales características epidemiológicas y la tasa de embarazo espontáneo en mujeres con esterilidad y lesiones cervicales con o sin la infección por VPH. Se incluyeron 61 pacientes evaluados con un protocolo diagnóstico por Esterilidad, enviadas para valoración a la Clínica de Colposcopia, por presentar alguna lesión cervical. En el grupo I VPH positivo (n=45), lesiones acetoblancas con pérdida parcial de epitelio y en grupo II, VPH negativo (n=16), lesiones por Cervicitis crónica, con eversión glandular, metaplasia inmadura y pérdida parcial del epitelio. Asimismo se solicitó citología cervicovaginal, colposcopia y biopsia dirigida. Las pacientes fueron sometidas al tratamiento médico convencional de estas lesiones. Cada paciente fue seguida por un año para evaluar la tasa de embarazo espontáneo o asociada al tratamiento médico de esterilidad. El diseño del estudio fue retrospectivo, descriptivo con una cohorte histórica de 1991 a 1996. El promedio de tiempo de esterilidad fue 4.86 en el grupo I y de 3.5 en el grupo II. Se observó una tasa de embarazo de 35,55 por ciento (16/45) en las pacientes con VPH+ y de 37.5 por ciento (6/16) en las pacientes con lesiones cervicales sin VPH. De estas pacientes, el 75 y 66 por ciento fueron embarazos espontáneos y el 25 y 33.3 por ciento con tratamientos como citrato de clomifeno y hMG, en los grupos I y II respectivamente. La media de tiempo para el embarazo espontáneo postratamiento fue de 8.87 y 7.16 meses respectivamente. La frecuencia del factor tuboperitoneal (exceptuando lesiones endometriósicas) fue de 53.57 por ciento y 46.66 porciento respectivamente. En las pacientes con lesiones producidas por VPH, la infección genital que sospecha por citología cérvico-vaginal en menos de 25 por ciento de los casos. Este estudio muestra nuevamente, que en mujeres con esterilidad y lesiones cervicales hay que sospechar la presencia de infecciones por VPH...