Detection of human papillomavirus in organs of upper genital tract in women with cervical cancer

In this study, we evaluated the presence of human papillomavirus (HPV) DNA in organs of the female upper genital tract, using nine hysterectomy and salpingo-oophorectomy specimens affected by HPV-positive invasive cervical carcinomas, to establish if cervical HPV infection can spread to upper tracts of the female genital system. HPV DNA was evaluated by polymerase chain reaction (PCR) in all cervical carcinomas as well as in all tracts of the genital system. Then, these data were compared with the results obtained from PCR study of five other hysterectomy and salpingo-oophorectomy specimens (control cases). The criteria used for selection of the control cases were informed consent of the patients for research at the time of surgery, absence of neoplasms, absence of any anatomic lesion caused by HPV in cervix, and external genitalia. All selected cases were squamous cervical carcinomas. PCR analysis revealed HPV DNA in all cases of cervical carcinoma. The HPV DNA was detected as weak positivity on PCR analysis in other organs of the genital system. However, the distribution of HPV DNA varied in the various cases and in the different tracts of the same hysterectomy and salpingo-oophorectomy specimen. We believe that the HPV DNA, detected as a weakly positive signal, in the upper genital tract of patients who have a cervical squamous carcinoma could be a reflection of a latent HPV infection, as well as a sign of the existence of micrometastases containing HPV DNA, which cannot be detected by conventional histologic techniques.     

Human papillomavirus in the oral mucosa of women with genital human papillomavirus lesions

OBJECTIVES: Human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases worldwide. We determined the frequency of HPV in the oral cavity of women with and without genital HPV lesions. MATERIAL AND METHODS: All patients were seen at the Department of Gynecology, Women's Health Center and the State University of Campinas, Sao Paulo, Brazil and submitted to a general physical and gynecological examination plus an evaluation of the oral cavity. Detailed histories investigated their sexual practices. HPV in the oral cavity was determined by polymerase chain reaction using consensus primers in 70 women presenting with histopathology-confirmed clinical HPV lesions in the genital region and 70 women negative by gynecological, colposcopic and cytological examination for clinical or subclinical HPV lesions. RESULTS: Oral HPV was detected in 29 (20.7%) of the subjects. Among the positive women, 26 (89.7%) were also positive for genital HPV as opposed to only 3 (2.7%) who were genital HPV-negative (p < 0.0001). The overall prevalence of HPV in the oral cavity of patients with and without genital HPV was 37.1 and 4.3%, respectively, (p < 0.0001). The presence of oral HPV was unrelated to the practice of fellatio (22% versus 19%). CONCLUSION: Patients with HPV genital infection have a greater frequency of HPV in their oral mucosa.     

The treatment of human papillomavirus lesions of the lower genital tract

Anogenital warts are caused by human papillomavirus types 6 and 11 infection, and are the most common clinical manifestation of lower genital tract human papillomavirus infection. They are the most frequently recognized sexually transmitted disease in the developed world, occurring at an incidence rate of 0.5-1.2% in young men and women aged 18-25 years. Many therapies for genital warts have been described, none being ideal in that all therapeutic modalities fail to clear the warts in a proportion of patients, and the recurrence of warts after successful treatment is seen with all treatments. The current knowledge base of the therapy of genital warts is flawed by a lack of good natural history data either with treatment or without treatment over longer periods of time, in that most trials report comparisons of monotherapies over a short time and there is a lack of structured trials addressing consecutive therapies over longer durations, as occurs in real-life clinical situations.     

Correlation of histology with human papillomavirus DNA detection in the female genital tract

Genital condyloma and intraepithelial neoplasia secondary to Human Papillomavirus (HPV) infection are characterized by perinuclear halos and marked nuclear atypia (koilocytotic atypia) on cytologic and histologic examination. However, at times the histologic findings, including the degree of nuclear atypia, may be suggestive but not absolutely diagnostic of an HPV related neoplasm. HPV DNA sequences were detected in 63 and 56% of colposcopically visible vaginal and cervical lesions, respectively, that were diagnosed as condyloma or intraepithelial neoplasia. HPV DNA sequences were detected in 14 and 47% of vaginal and cervical lesions, respectively, that did not fulfill the histologic criteria of condyloma or intraepithelial neoplasia (i.e., "nondiagnostic"). When examining cervices from patients with no visible lesion and no recent history of an abnormal pap smear, 5.5% had detectable HPV DNA sequences. The histologic findings in this group were equivalent to the virus-negative cases and similar to the "nondiagnostic" cervical lesions. These findings suggest that the detection rate of HPV DNA in "nondiagnostic" tissues is dependent on the site and presence or absence of a visible lesion. The rate is similar in cervical lesions regardless of the histologic findings whereas it is less in vaginal lesions when the histologic criteria of condyloma or intraepithelial neoplasia are not detected.     

In situ hybridization analysis of human papillomavirus DNA segregation patterns in lesions of the female genital tract

Various histologic features may be used to divide human papillomavirus (HPV)-related lesions of the genital tract into two groups: condylomata and "low-grade" or grade 1 cervical intraepithelial neoplasias (CIN 1) versus "high-grade" or grade 2 and 3 intraepithelial neoplasias. Using in situ hybridization analysis we correlated HPV DNA type with histologic features in 350 biopsies of lesions from the cervix, vulva, and perianal region. HPV DNA was most commonly found in vulvar and perianal condylomata (39/46, 85%), whereas the rate in CIN 1 lesions was 72% (86/120). The rates were 53% (40/76) and 57% (12/21) in CIN 2/3 and vulvar intraepithelial neoplasm (VIN) grades 2 and 3, respectively. The HPV type in all but 2 of the 39 perianal and vulvar condylomata which contained HPV was 6/11. Despite their similar histologic features, the HPV type in only 23 of 86 (27%) CIN 1 cases with detectable HPV was 6/11 compared to 31 of 86 (36%) which contained HPV 16-related DNA and 32 of 86 (37%) which contained HPV 31,-33, or -35-related DNA. The viral DNA in the majority of CIN 2/3 lesions and all of the VIN 2/3 lesions was HPV-16 related; no CIN 2/3 or VIN 2/3 lesion had HPV 6/11-related DNA. It is concluded that although cutaneous genital tract condylomata are highly associated with HPVs of low oncogenic potential (types 6 and 11), these HPV types are not as frequent as the oncogenic HPVs (16, 31, 33, and 35) in CIN 1 lesions. Further, HPV 6/11 appears to be very rarely associated with CIN 2/3 or VIN 2/3 lesions.     

Immune responses to human papillomavirus in genital tract of women with cervical cancer

OBJECTIVES: To address a question whether immune responses to HPV infection play a role in control of cervical cancer, we analyzed systemic and mucosal immune responses to HPV in women who underwent radical hysterectomy for cervical cancer (HCC) or loop conization due to cervical dysplasia (LOOP), or had hysterectomy for other reasons (HNN). METHODS: HPV-specific antibodies in sera and vaginal washes were determined by ELISA using recombinant HPV 16 E7 oncoprotein. Cytokines in vaginal washes were assayed by Linco cytokine multiplex method using Luminex technology. Differential gene expression profiling in cervical tumor was determined by microarray analysis and Real-time RT-PCR. RESULTS: While levels of HPV-16 E7-specific IgG in vaginal wash were significantly higher in women undergoing HCC and HNN, the levels of the HPV-16 E7-specific IgA in vaginal wash of women with cervical cancer and cervical dysplasia were lower as compared to patients in HNN. Proinflammatory cytokines, such as IL-6 and IL-8, were dominant in vaginal washes of all subjects studied. However, no pattern of Th1-type and Th2-type cytokine induction was observed as demonstrated by protein analysis as well as differential gene expression profiling in cervical tumor. CONCLUSIONS: These results suggest a selective down-regulation of local HPV-specific IgA responses in women with cervical cancer.     

High prevalence of oncogenic human papillomavirus in the genital tract of women with human immunodeficiency virus.

OBJECTIVE: The goal of this study was to determine the prevalence of human papillomavirus (HPV) and squamous intraepithelial lesions (SILs) in women infected with human immunodeficiency virus (HIV) in Mexico. METHODS: Cases included women who were positive for human immunodeficiency virus (HIV) and accepted to participate. There were two control groups in this study: group A, heterosexual partners of HIV+ men; group B, commercial sex workers. Gynecologic examination was performed in all participants. Also, a cervical smear with colposcopy and a sample for detection of HPV DNA by polymerase chain reaction (PCR) were obtained in all subjects, as were CD4+ counts. Relative risks (RR) and 95% confidence interval were calculated. RESULTS: Eighty-five HIV+ women agreed to participate in this study; the route of HIV infection was heterosexual in 78.8%; transfusion in 8.2%; paid donors in 3.5%; and 9.4% unknown. A total of 9 controls were included: 4 from group A and 5 from group B. HPV DNA was detected by PCR in 57 (69%) cases and in 26 (29%) controls from both groups (P < 0.0001). The RR of HPV infection was 5.5 (2.7-11.5). Also, a significant difference in the prevalence of high-risk HPV types was observed between cases and controls, RR = 12.8 (4.07-42.9). These associations were independent of CD4+ counts and antiretroviral therapy. No association was observed between HIV infection and the risk for high-grade SIL. CONCLUSIONS: We observed a high prevalence of oncogenic HPV types in HIV-positive women. These women should be screened regularly for early diagnosis of premalignant lesions and prevention of cervical cancer.     

Detection of human papillomavirus DNA in biopsy-proven cervical squamous intraepithelial lesions in pregnant women.

This study analyzed cervical squamous intraepithelial lesions (SILs) in pregnant women for human papillomavirus (HPV) using in situ hybridization analysis. HPV DNA was detected in 77% (23/30) of low-grade SILs as compared to 89% of such lesions in nonpregnant women. The detection rate in high-grade SILs was 41% (9/22) compared to 70% in nonpregnant women. Analysis by the polymerase chain reaction showed that the detection rates were similar (96-100%) for low- and high-grade lesions in pregnant and nonpregnant women, which demonstrates that in situ negative tissues indeed contained HPV DNA. Low-grade SILs contained a heterogeneous group of at least 14 different HPV types, whereas most high-grade SILs contained HPV 16. We concluded that cervical SILs in pregnant women are invariably associated with HPV. Further study is needed to determine which of several possible variables, such as the age of the lesion and the viral copy number, may explain the apparent decreased detection rate of HPV by in situ hybridization in SILs during pregnancy.     

Human papillomavirus DNA and mRNA positivity of the anal canal in women with lower genital tract HPV lesions: predictors and clinical implications

Objective

Women with HPV related pathology of the lower genital tract are at higher risk for AIN and anal cancer than the general population. A strategy to identify anal disease in these women has not been formulated. The aim of this study is to examine the feasibility of HPV related biomarker testing on anal smears, to identify the risk factors for anal HPV positivity and to provide information of the clinical implications of anal HPV infection in this population.

Methods

In women referred for colposcopy because of HPV related pathology of the lower genital tract (cervical cancer, CIN, VIN, warts) a detailed questionnaire, an anal smear and a cervical smear were taken. On each sample morphological cytology, flow cytometric evaluation of E6&7 mRNA, and HPV DNA detection and typing were performed. Women with a positive anal result were referred for high resolution anoscopy.

Results

So far 235 women have been included (mean age 34.3). HPV DNA, high-risk HPV DNA, high-risk mRNA was detected in 45%, 31% and 8% of the anal smears and in 56%, 39% and 25% of the cervical smears respectively. Absolute or partial concordance of the types between the cervix and the anus was seen in 74%. Positivity for mRNA was significantly lower in the anus than the cervix (8% vs 25%). Logistic regression analysis revealed risk factors for the presence of anal HPV DNA (> 3 lifetime sexual partners and presence of cervical HPV DNA), hr HPV DNA (presence of cervical hr HPV DNA), and hr mRNA (presence of cervical hr mRNA). Twelve months after LLETZ 53% of women were cervical HPV negative, but 25% of those were still HPV positive in the anus.

Conclusions

HPV infection of the anus is common in this group and is interlinked with the cervical infection. Anal HPV E6&7 mRNA expression is less common than in the cervix. Possible clinical implications of anal infection could be the development of AIN and recurrence of CIN after treatment due to cervical reinfection from the anal reservoir. The use of HPV biomarkers is feasible in anal smears, although especially DNA testing as triage method for referral to anoscopy is probably inappropriate due to high positivity rate.     

2285例女性下生殖道人乳头状瘤病毒感染筛查结果分析

目的探讨门诊就诊的高危人群人乳头状瘤病毒(HPV)感染的型别分布和年龄特征.方法 2004-10-2005-11采用核酸分子快速杂交分型技术,对2285例于北京大学第一医院妇产科门诊就诊妇女进行生殖道21种HPV亚型感染筛查.结果 HPV总感染率57.1%(1305/2285),21种型别均被检出,感染率最高的型别是HPV16,其他常见型别依次为58、52、53、33、6、CP8304、11、31和18型,高危型感染41.0%,低危型7.2%,多型感染8.9%.2285例HPV感染高峰年龄在30～34岁(11.3%),40～44岁年龄段下降到7.6%,各年龄段HPV感染检出率差异有显著性意义(P＜0.01).结论门诊高危人群HPV感染率57.1%,常见型别为HPV16、58、52、53、33、6、CP8304、11、31和18型;人群感染高峰在30～34岁年龄段.     

Screening for human papillomavirus infections of the lower genital tract

Human papillomavirus (HPV) infections are the etiologic agents in the development of lower genital tract neoplasia. Risk factors for the development of cervical cancer include high risk HPV subtype, persistent infection, high viral load, immunosuppression, tobacco use, and absence of cytologic screening. Strategies to increase the yield on screening for cervical neoplasia have included the use of HPV subtyping. This review summarizes the studies of HPV testing as a primary screening test, an adjunct for screening low and high risk populations, and for follow-up in women with persistent squamous intraepithelial lesions. Use of HPV screening is not yet applicable to populations with successful screening programs in place. Molecular markers for progression to cancer in women with persistent high risk HPV infections need to be discovered.     

Prevalence of genital human papillomavirus among Lebanese women

PURPOSE: The purpose of this study was to determine the prevalence of human papillomavirus (HPV), and more specifically of HPV 16, in a group of Lebanese women. MATERIALS AND METHODS: Type-specific prevalence of cervical HPV and the presence of cytological abnormalities were determined in a cohort of Lebanese women. The population included 1,026 women, 18-76 years, seeking routine gynecological care at a tertiary care center. Demographic and behavioral data were collected. HPV DNA was detected in cervical scrapes by polymerase chain reaction using consensus primers. Cervical cytological abnormalities were identified by Papanicoleau (Pap) smears. RESULTS: The mean age of our population was 40 +/- 11.3 years. General HPV DNA was detected in 50 patients (4.9%). The high-risk HPV type 16 DNA was detected in 31 patients (3%). Patients with HPV 16 were more likely to have an abnormal pap smear than those with negative tests (6.6% vs 1.6%, p < 0.05), and more likely, but not significantly, to be smokers (21.4% vs 18.4%, p = 0.5). The age-specific prevalence of HPV increased with age and peaked at 60-69 years. CONCLUSIONS: The prevalence of HPV in this small group of Lebanese women is similar to its prevalence in the Mediterranean countries. The presence of HPV, its known association with the development of cervical neoplasia, and the lack of a universal screening program for cervical cancer in our country should be used to enforce implementation of proper screening programs.     

Clinical findings among young women with genital human papillomavirus infection

OBJECTIVE: The purpose of this study was to identify clinical signs and symptoms associated with detection of human papillomavirus (HPV) DNA in the female genital tract. STUDY DESIGN: A total of 516 university students (18 to 24 years old) enrolled in a cohort study that included the collection of genital specimens for HPV DNA testing every 4 months for up to 4 years. Reported symptoms and objective clinical findings of women with and without HPV DNA were compared by multivariate analysis. RESULTS: Acute and persisting HPV infections were not associated with discharge, itching, burning, soreness, or fissures. Clinical evidence of genital warts was statistically associated only with HPV types 6 and 11. Detection of any HPV DNA was associated with bacterial vaginosis (BV). Furthermore, a time lag analysis suggests that HPV infection usually precedes detection of BV. CONCLUSION: Most women who acquire genital HPV infection are asymptomatic; some, however, are at increased risk for BV.     

Immunology of human papillomavirus infection in lower genital tract neoplasia

Despite its being a relatively common virus, the study of human papillomavirus infection has lagged behind that of other viruses. Human papillomaviruses do not provoke strong systemic antibody or T-cell responses. Furthermore, the majority of those infected do not display clinical symptoms and are able to clear the virus by unknown mechanisms. In the last decade, however, research into human papillomavirus immunology has blossomed, for two main reasons. First, there is strong circumstantial evidence that the immune system can control papillomavirus infection, since the prevalence of human papillomavirus-associated neoplasia is increased in immunocompromised individuals. Second, the strong association between human papillomavirus infection and cervical cancer has led to attempts to develop prophylactic or therapeutic vaccines. In this chapter, our current knowledge of human papillomavirus immune responses will be reviewed, and how this relates to clinical practice will be discussed.     

A comparison of slot blot, southern blot, and in situ hybridization analyses for human papillomavirus DNA in genital tract lesions

Genital tract lesions were analyzed for human papillomavirus (HPV) DNA by in situ hybridization using probes of HPVs 6/11, 16/18, and 31/33/35. All of the HPVs detected in vulvar and perianal condylomata by in situ hybridization were HPV 6/11-related, whereas the majority of HPVs detected in cervical intraepithelial grade I lesions were types 16/18- and 31/33/35-related. None of the lesions with histologic features equivocal for HPV infection had detectable HPV DNA by in situ hybridization, though some did contain HPV DNA sequences as ascertained by filter hybridization analysis. The sensitivity of in situ hybridization was compared with that of filter hybridization (slot blot and/or Southern blot). The correlation was high (28 of 30) for cases that contained HPVs 6/11 or 16, as deduced by filter hybridization, and was much less (ten of 29) for cases that contained HPVs distinct from types in the filter hybridization probe cocktail (HPVs 6/11, 16, 18, 31, 35 and 51). There was a high concordance between the results of Southern blot hybridization and slot blot hybridization analyses, especially with cases that contained HPVs 6/11 and 16. In situ hybridization, slot blot, and Southern blot hybridization analyses are all very effective in detecting the common HPV types (HPVs 6/11 or 16). In situ hybridization is useful in differentiating cervical lesions that contain HPV 6/11 from those that contain HPV 16 or other types with oncogenic potential. However, filter hybridization is superior to in situ hybridization when analyzing cases with histologic findings equivocal for HPV infection or cases that contain HPV types related to, but distinct from, the types included in the probe.     

Oral condyloma lesions in patients with extensive genital human papillomavirus infection.

OBJECTIVES: The incidence, location, and morphologic appearances of human papillomavirus oral lesions in patients with genital condylomatosis were investigated with clinical, colposcopic, and histologic examination as diagnostic procedures. The human papillomavirus types were also evaluated with filter in situ hybridization. STUDY DESIGN: One hundred one patients, 66 female and 35 male, with genital condyloma underwent an oral cavity examination. Ninety-nine (99%) practiced orogenital sex, and all were asymptomatic for oral lesions. RESULTS: Ninety-one underwent biopsy; histologic studies gave a diagnosis of condyloma in 48% of 101 specimens collected. In 8 of 91 (9%) oral lesions were suspected by naked-eye examination; they were confirmed histologically in all eight. Of 83 patients suspected of having oral condyloma on colposcopic examination, 38 (46%) were confirmed histologically. Thus 38 of 46 patients (83%) had oral condyloma not visible to the naked eye. Colposcopically, oral lesions appeared filiform (50%), moruloid (26%), and mixed (24%). Twenty cytologic oral samples were also collected for deoxyribonucleic filter in situ hybridization analysis. Human papillomavirus deoxyribonucleic genital types were observed in 45% (9/20) of all oral scrapings collected, and all were histologically confirmed. CONCLUSION: Our data indicate that genital human papillomavirus types are capable of establishing a local infection in the oral cavity and demonstrate a high incidence of human papillomavirus oral lesions in patients with genital condyloma.     

Serum retinol level in patients with pre and malignant lesions of female genital tract

The blood retinol level in women with genital tract malignancies and in a control group has been determined. In all groups of patients the mean serum retinol level was significantly lower than in control group. Based on these results we can postulate, that the low retinol level could predispose to the development of malignancy in female genital tract.     

Detection of human papillomavirus DNA in cervical lesions by in situ hybridization using biotinylated DNA probes

In situ hybridization (ISH) for human papillomavirus (HPV)-6, -11, -16, -18, and -31 DNA was performed on 615 formalin-fixed paraffin-embedded cervical biopsies using biotinylated DNA probes. Results were obtained from 584 samples with 266 (45.5%) containing HPV-DNA sequences. Ninety percent of condyloma acuminatum specimens were positive for HPV-DNA with 18 of 19 positive cases containing HPV-6 or -11 DNA. The detection rate of HPV in cervical intraepithelial neoplasia (CIN) lesions was 50.6% (239 of 472), while only 8 of 91 (8.9%) cervical biopsies considered to be histologically normal or with minimal dysplasia contained HPV-DNA as demonstrated by ISH. The prevalence of HPV-16, -18, and/or -31 DNA increased with the severity of the lesions, with 20 of 20 (100%) positive CIN-III lesions containing these viral types compared with 102 of 157 (65.0%) positive CIN-I lesions. ISH with biotinylated DNA probes appears helpful in identifying lesions containing higher risk viral strains.