HPV and other risk factors of oral cavity/oropharyngeal cancer in the Czech Republic.

OBJECTIVE: An association between high-risk human papillomavirus (HR HPV) infection and a risk of development of a subgroup of head and neck cancers has been proposed recently. The main risk factors of oral and oropharyngal cancer observed in our population are smoking and alcohol consumption. The incidence of oral/oropharyngeal tumours in the Czech Republic is relatively high and there are no data available about the prevalence of HPV DNA presence in these tumours. MATERIALS AND METHODS: Eighty patients with a primary oropharyngeal cancer were enrolled. The presence of HPV DNA has been evaluated by polymerase chain reaction in 68 cases from which the tumour tissue and demographical and clinical data were available. The typing of HPV was performed by nucleotide DNA sequencing. RESULTS: The HPV DNA was detected in 51.5% of samples tested. Among the HPV DNA positive tumours, 80% contained HPV16. In the analysed group there were 54 men and 14 women. The prevalence of HPV DNA was lower in oral (25%) than in oropharyngeal (57%) tumours, and higher in never smokers (100%) and never drinkers (68.8%). HPV DNA presence was not related to gender, age, number of lifetime sexual partners or practice of oral-genital sex, size of tumour or presence of regional metastases. CONCLUSIONS: The difference in the prevalence of HPV DNA positive tumours between cases of oral cavity and oropharyngeal carcinoma exposed and not exposed to tobacco or alcohol support the theory that HPV DNA positive tumours form an aetiologically distinct subgroup of head and neck tumours.

Persistent HPV infection in postmenopausal age women.

OBJECTIVE: Persistence of human papillomavirus (HPV) is associated with an increased risk of developing cervical SIL and cancer in young women. Because this association in older, postmenopausal age women has received little attention, we evaluated persistence of HPV among women in this age group. METHODS: Women (n=105) ages 45-64 were examined annually for 7 years to evaluate HPV in cervical cytologic specimens. PCR, dot blot hybridization and DNA sequencing were used to detect HPV types. RESULTS: The cumulative prevalence of HPV was 34%, and 24% had HPV high-risk oncogenic types which are associated with genital cancers. The most common oncogenic types were HPV-16 (72%) and HPV-31 (16%). The persistence rate of HPV infection was 16%. No specific risk factors were associated with repeat viral positivity. CONCLUSION: Postmenopausal women are infected with persistent oncogenic HPV at a substantial rate, supporting the need for continued screening in postmenopausal women to detect preneoplastic genital lesions.

[HPV-associated tonsillar cancer. An update]. / HPV-assoziierte Tonsillenkarzinome. Ein Update.

The major risk factors for head and neck squamous cell carcinomas (HNSCC) are considered to be tobacco and alcohol. A link between oncogenic types of the human papilloma virus (HPV) and the risk of HNSCC has been suggested in the literature. However, the causal link is now becoming more firmly established on the basis of recent analyses. About 20% of all HNSCC and more than 50% of tonsillar cancers contain HR-HPV. The causal role of HPV-infection in carcinogenesis and the molecular mechanisms involved could thus far be best elucidated in the case of cervical carcinomas. New insights and increasing evidence for the analogy of HPV-positive HNSCC with cervical cancer are discussed. The definition of HPV-positive HNSCC has become more important due to the implications for risk factors and prognosis.

HPV prevalence and concordance in the cervix and oral cavity of pregnant women.

OBJECTIVES: This investigation examined human papillomavirus (HPV) in pregnant women in order to characterize viral prevalence, types and concordance between infection in the cervix and in the oral cavity. METHODS: A total of 577 pregnant women seeking routine obstetric care were evaluated for HPV infection in their cervix during gestation and immediately before delivery, and in the oral cavity during gestation. Male partners present during the gestational clinic visit also provided a specimen from their oral cavity. HPV assessment was performed by PCR, dot blot hybridization and DNA sequencing. A sexual and health questionnaire was completed by the pregnant women. RESULTS: HPV prevalence in women was 29% in the cervix and 2.4% in the oral cavity. Among those with both gestational and delivery specimens, 35% were infected at least once and 20% had infection at both intervals. At delivery, 68% of infected women had an oncogenic HPV type in the cervix. There was no type-specific HPV concordance between the two cervical specimens, nor cervical and oral results in women, nor with cervical and oral findings between partners. CONCLUSION: The lack of association in HPV positivity and types between the cervix and oral cavity in these women suggests that self-inoculation is uncommon. This source of infection does not appear to be from oral contact with a current male partner, since there also was no concordance between partners. These results suggest either other modes of HPV transmission or differences in susceptibility to HPV infection or its clearance in the oral cavity and genital mucosa.

Is use of hormone replacement therapy associated with increased detection of human papillomavirus and potential risk of HPV-related genital cancers?

Oral contraceptives (OC) are a risk factor for female genital cancers and in vivo studies have shown that progestins stimulate human papillomavirus (HPV) gene expression. A similar role for hormone replacement therapy (HRT) has received little evaluation. Cervical/vaginal specimens were obtained to detect HPV from postmenopausal women (n = 429) seeking annual gynaecologic care. HPV was detected in 14% of women and 4.4% had high-risk, oncogenic types. HPV prevalence was similar across current, past and never HRT users. After adjustment for HPV-related risk factors, current and past user status showed no increased viral detection compared with never users. HRT duration also did not elevate risk among current users. However, longer duration (adj. OR 1.5/year, 95% CI 1.0-2.3) and longer latency (adj. OR 1.2/year, 95% CI 0.9-1.7) among past users of oestrogen/progestin regimens were associated with greater risk. Overall use of HRTs was not associated with HPV detection or disease. However, past users of combination HRTs had significantly greater risk of HPV detection with longer HRT duration and latency, similar to OC-HPV findings. The recommendation that postmenopausal women continue HRTs long term may lead to an increased development of HPV-related diseases, of particular concern among those who discontinue HRTs and subsequent gynaecologic care for early cancer detection.

Human papillomavirus in the oral cavities of children and adolescents.

OBJECTIVE: The purpose of this pilot study was to determine the frequency of human papillomavirus (HPV) in the oral cavities of children and adolescents and to identify potential risk factors for HPV infection. STUDY DESIGN: Sociodemographic information was obtained on 268 healthy infants, children, and adolescents who were < or = 20 years old. Oral squamous cells were collected from swabs with young children and from oral saline solution rinses with older children and adolescents. Extracted DNA was evaluated for HPV by polymerase chain reaction, dot blot hybridization, and DNA sequencing. Factors associated with the presence of HPV were tested by using chi(2), Fisher's exact test, and logistic regression tests. RESULTS: HPV was detected in 6.0% of the participants. HPV frequency among young children (<7 years old) was 8.7% (11/127), and among adolescents (13-20 years old) it was 5.2% (5/97). HPV was not detected in children aged 7 to 12 years old (0/44). Fifty-four percent (6/11) of HPV-positive children were 1 year of age or less; 3 of the HPV-positive children (<7 years old) were delivered by cesarean section. No statistically significant association was found between the detection of HPV in the oral cavity and method of delivery or gender; parent's race, education, HPV-related conditions, smoking history, or number of sex partners; or adolescent's smoking history or history of sexual activity. CONCLUSIONS: This study suggests that HPV is present in the oral cavity primarily in children 2 years old and younger and in adolescents 13 years and older. Cesarean delivery was not protective against oral HPV infection; in fact, half of the HPV-positive infants were born by cesarean delivery.

Human papillomavirus and risk of laryngeal cancer.

We determined the relationship between human papillomavirus (HPV) infection and the HPV types detected in 44 patients with squamous cell carcinoma, 10 laryngeal leukoplakia patients, and 12 patients evaluated for benign laryngeal conditions (controls). The sources of HPV DNA were from brushings from the upper respiratory tract and lesion (benign or malignant), oral rinses, and biopsies of patient lesions. Polymerase chain reaction (PCR) and DNA sequencing were used to identify and type HPV. We detected HPV in 25.0% (11/44) of patients with laryngeal cancer, in 30.0% (3/10) of patients with laryngeal leukoplakia, and in 16.7% (2/12) of noncancer controls. Patients with cancer were not more likely to be identified with oncogenic HPV types ( 18.2%) than either the leukoplakia group (20%) or the control group (16.7%). An increased risk of disease was associated with current tobacco use and former alcohol drinking in cancer patients versus controls and in leukoplakia patients versus controls (all p < .05). After we controlled for tobacco and alcohol effects on the risk of disease, exposure to oncogenic HPV types was associated with an increased risk of laryngeal cancer (odds ratio = 3.0) and of laryngeal leukoplakia (odds ratio = 6.0) compared to controls, although the results were not statistically significant. This study suggests that although HPV infection and HPV oncogenic types are not found at a higher frequency in laryngeal cancer or laryngeal leukoplakia as compared to controls, infection is associated with an increased risk of disease after controlling for the effects of alcohol and tobacco use.

Use of hormone replacement therapy (Hrt) And detection of human papillomavirus (Hpv) Dna in postmenopausal women.

PURPOSE: This study investigated the association between detection of HPV DNA among current and past HRT users compared to never HRT users, duration of HRT use, and type of HRT (estrogen, estrogen/progestin).METHODS: Postmenopausal women (n = 390) were recruited from a university hospital and completed a questionnaire regarding 1) HRT use, duration, and type, 2) reproductive and sexual history, 3) smoking and alcohol use, and 4) HPV-related diseases. Cervical specimens were obtained for Pap smears, and for the presence of HPV using PCR/dot blot and DNA sequencing, and Southern blot or SSCP. Age-adjusted odds ratios (OR), 95% confidence intervals (CI), and logistic regression examined the association between HRT use and risk of HPV detection.RESULTS: The frequency of HPV was 10%, with 2.8% oncogenic types. Compared to Never Users, Current (adj. OR = 1.50, 95% CI = 0.55, 4.07) and Past (OR = 1.96, CI = 0.56, 6.86) HRT users had an elevated risk of HPV detection. Although HRT duration among Current Users was not statistically significant (OR = 1.01, 95% CI = 0.95, 1.07), duration among Past Users was significantly associated with an increased risk of HPV detection (OR = 1.30, CI = 1.06, 1.61). In addition, Past Users of estrogen/cyclic progestin HRT regimens were significantly more likely to be detected with HPV (OR = 1.82, CI = 1.11, 2.97) compared to Never Users.CONCLUSIONS: These findings indicate that risk of HPV detection is increased among users of HRTs, particularly among Past Users associated with longer duration and among those taking HRT regimens that included a progestin. Results suggest a latency or duration effect may be important in elevating the risk. They also support previous in vitro studies of progestin effects on HPV gene regulation.

p53 polymorphism, human papillomavirus infection in the oral cavity, and oral cancer.

OBJECTIVES: Human papillomavirus (HPV) infection has emerged as a risk factor in oral carcinogenesis. An arginine-coding polymorphism of the tumor suppressor protein p53 at codon 72 is more readily degraded by the HPV oncoprotein E6. Our objective was to evaluate the association between p53 polymorphism at codon 72 and HPV infection in the oral cavity, as well as its association with oral cancer. STUDY DESIGN: Oral squamous cells from 202 patients with oral cancer and 333 age-sex frequency matched controls were evaluated by polymerase chain reaction for the presence and type of HPV and for alleles of codon 72 in p53. Fisher exact test and chi(2) tests were used to evaluate the data. RESULTS: The p53 codon 72 polymorphism is not associated with HPV infection, whether comparing HPV-negative controls with HPV-positive controls or comparing HPV-negative cases with HPV-positive cases. Additionally, we found no association with the codon 72 polymorphism and oral cancer, whether comparing HPV-negative controls with HPV-negative cases or comparing HPV-positive controls with HPV-positive cases. CONCLUSIONS: There is no association between p53 codon 72 polymorphism and HPV infection or between the p53 polymorphism and the risk of oral cancer.

Resection of metastatic melanoma following wire localization guided by computed tomography or ultrasound.

Melanoma is an aggressive tumour with the propensity to metastasize through the lymphatic system and blood. Patients at high risk for developing metastatic disease are evaluated clinically together with chest X-rays and when indicated computed tomography (CT) scans. Wire localization is routinely used in non-palpable breast cancer to facilitate surgical resection. This study demonstrates the applicability of wire localization and surgical resection of non-palpable, deep subcutaneous or Intramuscular metastatic melanoma detected by CT or ultrasound. The medical records of six patients with malignant melanoma were retrospectively reviewed. Each patient with malignant melanoma developed metastatic involvement detected by CT scan or ultrasound at the UCSF/Mount Zion Medical Center, California, USA. A Copanz needle was inserted into the tumours under local anaesthesia. The patients were transported to the operating room and underwent wire-guided surgical resection of the tumour under general anaesthesia or intravenous sedation. In all six patients the tumour was successfully resected following CT- or ultrasound-guided wire localization of the metastatic foci. In conclusion, nonpalpable metastatic melanoma may be resected using CT-or ultrasound-guided wire localization. This technique offers several advantages, Including minimal surgical dissection, shorter operative times and decreased postoperative morbidity.

Human papillomavirus and risk of oral cancer.

Although human papillomavirus (HPV), a sexually transmitted virus, is established as a necessary cause for more than 95% of cervical carcinomas, the association with oral squamous cell carcinoma is less well delineated. The purpose of this study was to determine the frequency and types of HPV in squamous cells of a group of patients with newly diagnosed oral or pharyngeal cancer (n = 93) compared with an age- and gender-frequency-matched control group of patients with no history of oral cancer (n = 205). HPV was evaluated from a mouth rinse collection of cells in the oral cavity and tested by 32P-labeled HPV generic probes and DNA sequencing for HPV types. HPV was identified in 15% of the oral cancer cases but in fewer than 5% of the controls (P < .05). The risk of cancer associated with HPV infection was independent of tobacco and alcohol use (adjusted odds ratio [OR] = 3.70; 95% confidence interval [CI]: 1.47-9.32; P < .05). HPV types included similar and other types not identified previously in the genital tract. There was no statistically significant increased risk of cancer among former tobacco users (former vs. never users: adjusted OR = 0.67, 95% CI: 0.31-1.44, P < .05), but the risk was significantly increased for current users (current vs. never: adjusted OR = 2.63; 95% CI: 1.22-5.71; P < .05). Likewise, former alcohol users were not at increased risk of disease (former vs. never: adjusted OR = 1.78; 95% CI: 0.87-3.67), whereas current alcohol users were (current vs. never: adjusted OR = 2.57; 95% CI: 1.22-5.42; P < .05). HPV-related genital lesions (14.3% vs. 10.6%), oral-genital sexual behavior (42.4% vs. 45.2%), and number (11 or more) of sexual partners (23% v. 17%) were not significantly different between cases and controls. These data suggest that in addition to tobacco and alcohol, HPV plays a role in the development of oral cancer.

Serologic response to the E4, E6, and E7 proteins of human papillomavirus type 16 in pregnant women.

OBJECTIVE: In a seroepidemiologic study the effects of pregnancy and other factors on humoral response to human papillomavirus type 16 infection were examined. STUDY DESIGN: Multiple serum samples were taken at 3-month intervals for 15 months from 77 pregnant and 85 nonpregnant women. Serologic response to human papillomavirus type 16 proteins was analyzed with a peptide-based enzyme-linked immunosorbent assay. RESULTS: Seroreactivity was higher in nonpregnant women than in pregnant women, suggesting a reduced humoral immune response against human papillomavirus infections during pregnancy. Among the pregnant women a twofold to threefold decrease in mean reactivity in the E4 protein-based assay was detected between early gestation and delivery. The presence of human papillomavirus type 16 or 18 deoxyribonucleic acid was significantly associated with reactivity to the E6 protein (p = 0.0005) and the E4 protein (p = 0.06). Reactivity to the E4 protein also correlated with an abnormal Papanicolaou smear. CONCLUSIONS: The observation of changes in humoral response to genital human papillomavirus infections during pregnancy warrants further investigation with highly seroreactive assays.

The frequency of human papillomavirus detection in postmenopausal women on hormone replacement therapy.

Postmenopausal women enrolled in the Iowa portion of the postmenopausal estrogen/progestin interventions randomized clinical trial (n = 105) during 1989-1991 were studied for (i) the prevalence of human papillomavirus (HPV) in this older age population (ages 45-64), and (ii) the association between hormone replacement therapies (HRTs) and changes in detection of HPV over a 2-year time period. HPV is causative in most cervical and some other genital cancers and in the presence of steroid hormones has been shown to increase neoplastic transformation by HPV in vitro. Using PCR to detect HPV DNA, the overall frequency of the virus regardless of time period was 50.3% (n = 53) with a baseline (BL) frequency of 38.1% and the second year follow-up (FU) of 22.9%. The oncogenic types HPV-16 (75.5%) and HPV-31 (20.8%) were the most commonly reported. All those with persistently detected infection (10.5%), defined as HPV+ at both BL and FU, were identified with HPV-16 or -18. Between these two time periods there were no significant differences in HPV frequency between the placebo and combined HRT groups (BL-/FU+, 21% vs 18%; BL+/FU-, 71% vs 80%). While the study is based on a small sample, the findings suggest that short-term use of HRTs is not associated with an increased risk of HPV detection, but assessment of effects from long-term use is needed. The data also indicate that the frequency of HPV found in older women is higher than previously suspected but that short-term changes in HPV detected in this age group are unrelated to the development of precancerous cervical lesions.

Simple method for selective amplification of cDNA from a defined promoter.

A simplified technique for the detection of transcripts from a defined promoter is described. After reverse transcription, a PCR target sequence is selectively added to the 3' end of cDNA strands by DNA polymerase extension directed by an oligonucleotide template. Those cDNA molecules that do not have ends within a few nucleotides of the promoter start site are not extended and thus are excluded from subsequent amplification. Even when amplified products are visualized by ethidium bromide staining of agarose gels, this method requires only 1% of the RNA usually needed for detection of mRNA by standard RNase protection utilizing radiolabeled probes. In contrast to direct detection of cDNA by PCR, this procedure restricts amplification to a narrow subset of transcripts even when other overlapping colinear transcripts are present. We call this detection procedure specific amplification of cDNA ends (SPACE).

The genetic program of genital human papillomaviruses in infection and cancer.

Human papillomavirus (HPV) infection has been recognized as the major cause of cervical cancer. This article summarizes the functions of HPV gene products that cause abnormal cell growth--E6 and E7--and reviews how cellular and viral factors influence their synthesis. E6 and E7 inactivate two cellular tumor-suppressor gene products, p53 and RB. In cervical cancer, E6-E7 gene control is deranged by mutations in viral control sequences and in integrated HPV fragments by the disruption of the viral repressor E2. Elimination of this sequence makes E6-E7 mRNAs unstable, and deranges cellular regulation at the integration site. It is apparent that an intricate interplay of cellular and viral factors determines whether the outcome is active papillomavirus infection, viral latency, or ultimately, genital cancer.

Longitudinal study of the effects of pregnancy and other factors on detection of HPV.

OBJECTIVE: An epidemiologic study of 108 pregnant and 192 non-pregnant women was carried out to determine the effects of pregnancy and other factors on the detection of HPV infection, since both or larger numbers of pregnancies and HPV infection are known to be risk factors for cervical cancer. PATIENTS AND METHODS: Study participants were followed up at 3-months intervals for a period of 15 months (1-6 months after delivery). At each visit, two cervical specimens were taken, one for routine cytology and a second for HPV DNA hybridization using Virapap/Viratype, and a 5-ml blood sample was taken and a self-administered questionnaire was completed. RESULTS: In cervical specimens of the initial examination, HPV DNA was detected among 5.0% of pregnant and 5.2% of nonpregnant women, whereas HPV 16/18 was found in 80% of the HPV-positive specimens. Using multiple cervical specimens taken over time, 13.9% of the pregnant and 15.1% of the nonpregnant women tested positive for HPV DNA at least once. Adjusted for study group, age, and the number of available cervical specimens, ever detection of HPV infection (any type) was associated with more sexual partners and larger numbers of cigarettes smoked daily. An autoregressive generalized linear model was used to analyze the time-dependent multiple observations per study subject. Adjusting for age and trimester of pregnancy, the determinants of detecting high-risk HPV types (16/18 and 31/33/35) in the cervical specimen were an abnormal Pap smear, a positive HPV result in a preceding specimen, more than six sexual partners in the lifetime, and currently smoking more than 20 cigarettes per day with odds ratios of 10.9, 5.6, 3.2, and 2.7, respectively. CONCLUSION: Our data provide no evidence for a higher detection rate of HPV during pregnancy.

Perinatal transmission and maternal risks of human papillomavirus infection.

We conducted a prospective study to investigate whether human papillomavirus (HPV) could be vertically transmitted to neonates. Pregnant women (N = 203) were tested for HPV DNA infection during the third trimester and again during labor prior to delivery. Their newborns (N = 203) were tested 1 to 3 days after delivery. Among the mothers, 12.3% (N = 25/203) typed HPV positive at either or both maternal specimen collection periods, whereas only 1.0% of the neonates (N = 2/203) typed positive. This low transmission rate may be due in part to the fact that 65% of mothers who were HPV positive during the third trimester tested HPV negative by labor/delivery. The higher frequency of risks associated with maternal HPV infection were similar to those found in studies of cervical dysplasia and cancer: younger age at first intercourse and first pregnancy, number of sexual partners, and longer duration in use of oral contraceptives. In addition, those who were past smokers and had a shorter recency and latency period in smoking were more likely to be detected with HPV.

Human papillomavirus infection in papillomas and nondiseased respiratory sites of patients with recurrent respiratory papillomatosis using the polymerase chain reaction.

We examined human papillomavirus (HPV) infection in biopsy specimens and cellular scrapes that were taken from respiratory papillomas and six nondiseased sites from the respiratory tract of seven patients. Human papillomavirus was detected by polymerase chain reaction amplification, followed by DNA hybridization with probes for specific HPV types. All papillomas (100.0%, n = 5) were positive only for HPV type 6 or 11. In the nondiseased site specimens, 61.3% (19/31) of the specimens were positive, again only for HPV type 6 or 11. Among the nondiseased site specimens from the cervical trachea, intrathoracic trachea, and bronchus, 80% to 100% were HPV positive compared with only 25% to 50% of HPV infection detected in the nasopharynx, posterior tonsillar pillar, and aryepiglottic fold. These results support the tenet that HPV infection is present in clinically normal respiratory tract tissue and that the reservoir site of reinfection is more commonly in the lower airway. However, patients with upper-airway involvement were more likely to be diagnosed as having severe disease.

Transcriptional enhancer factor (TEF)-1 and its cell-specific co-activator activate human papillomavirus-16 E6 and E7 oncogene transcription in keratinocytes and cervical carcinoma cells.

The human papillomavirus (HPV)-16 oncogenes, E6 and E7, are transcribed preferentially in keratinocytes and cervical carcinoma cells due to a 5' enhancer. An abundant peptide binding to a 37 nt enhancer element was purified from human keratinocytes by sequence-specific DNA chromatography. This protein was identified as transcriptional enhancer factor (TEF)-1 by complex mobility, binding to wild-type and mutant SV40 and HPV-16 enhansons and antigenic reactivity with two anti-TEF-1 antibodies. TEF-1 is cell-specific, but its transactivation also depends on a limiting, cell-specific TEF-1 'co-activator'. We show that both TEF-1 and the TEF-1 co-activator are active in human keratinocytes and essential for HPV-16 transcription. TEF-1 binding in vivo was necessary for HPV-16 P97 promoter activity. Excess TEF-1 and chimeric GAL4-TEF-1 specifically inhibited the P97 promoter by 'squelching', indicating that HPV-16 transcription also requires a limiting TEF-1 co-activator. TEF-1 and the TEF-1 co-activator functions mirrored HPV-16 transcription by their presence in keratinocytes and cervical carcinoma cells and their absence from lymphoid B-cells, but also functioned in liver cells where the HPV-16 promoter is inactive. TEF-1 and its associated co-activator are thus part of a complex mechanism which determines the restricted cell range of the HPV-16 E6 and E7 oncogene promoter.