Broad Spectrum Of Human Papillomavirus Types Research Articles

Human papillomavirus 16L1-58L2 chimeric virus-like particles elicit durable neutralizing antibody responses against a broad-spectrum of human papillomavirus types.

The neutralizing antibodies elicited by human papillomavirus (HPV) major capsid protein L1 virus-like particle (VLP)-based vaccines are largely type-specific. An HPV vaccine inducing cross-neutralizing antibodies broadly will be cost-effective and of great value. To this end, we constructed HPV16L1-58L2 chimeric VLP (cVLP) by displaying HPV58 L2 aa.16-37 on the DE surface region of HPV16 L1. We found that vaccination with the HPV16L1-58L2 cVLP formulated with alum plus monophosphoryl lipid A (Alum-MPL) adjuvant elicited robust neutralizing antibodies in both mice and rabbits against all tested HPV types including HPV16/31/33/35/52/58 (genus α9), HPV18/39/45/59/68 (genus α7), HPV6/11 (genus α10), HPV2/27/57 (genus α4), and HPV5 (genus β1). Importantly, the cross-neutralizing antibody response was maintained at least 82 weeks in mice or 42 weeks in rabbits, and complete protection against HPV58 was observed at week 85 in mice. Our data demonstrate that HPV16L1-58L2 cVLP is an excellent pan-HPV vaccine candidate.

Detection of Merkel cell polyomavirus and human papillomavirus DNA in porocarcinoma

BackgroundIncreasing evidences support the role of Merkel cell polyomavirus (MCPyV) and human papillomavirus (HPV) in non-cutaneous and cutaneous tumours. Porocarcinoma is a rare malignant neoplasm that arises from the intraepidermal ductal portion of the eccrine sweat glands. The aetiology of porocarcinoma is largely unknown and no systematic studies have been done to investigate the implication of infectious agents in the pathogenesis of this tumour. ObjectivesTo investigate the possible association between MCPyV and/or HPV infection and porocarcinoma. Study designForty-four formalin-fixed paraffin-embedded (FFPE) porocarcinomas (40 primary and 4 metastatic) and 10 healthy skin specimens (controls), were analysed for the presence of MCPyV and HPV DNA using molecular detection methods. ResultsMCPyV DNA was found in 27/40 (68%) primary porocarcinomas and in 3/10 (30%) controls (Fisher exact test: p<0.04). No significant difference in viral load was observed between tumours and healthy skin. Moreover, 2/40 primary porocarcinomas tested positive for high-risk HPV16. Cutaneous beta-HPV infection was detected in 16/40 (40%) porocarcinomas and in 6/10 (60%) controls. No particular beta-HPV types were significantly associated with tumour or with healthy skin. Two out of 4 metastatic biopsies were MCPyV DNA positive. All metastatic samples had mixed infections with cutaneous HPV types. ConclusionsThis study demonstrated a significantly high prevalence of MCPyV and the presence of a broad spectrum of HPV types in porocarcinoma and provided the first available data about viral infections in this tumour. To understand the role, if any, of viral infections in the pathogenesis of porocarcinoma further studies are needed.

Comparison of Human Papillomavirus Detection and Genotyping with Four Different Prime Sets by PCR-Sequencing

ObjectiveTo assess and compare the Human Papillomavirus (HPV) detection efficiency and the potential clinical utility of PCR sequencing-based technology. MethodsFour HPV consensus primer sets (GP5+/6+, MGP, MY09/11, and PGMY09/11) were used in order to amplify a broad spectrum of HPV types for HPV infection in 325 cervical samples and the PCR products were sequenced afterwards for the HPV genotyping. ResultsThe HPV-positive rate was 75.4%, of which 35.5% harbored more than one HPV genotype. A total of 36 different genotypes was found, with HPV 16 (24.1%) being the most prevalent, followed by HPV 58 (13.3%) and HPV 52 (9.6%). There were substantial to almost perfect agreements between different primer sets regarding HPV detection efficiency, with the kappa value varying from 0.751 to 0.925, MGP, and PGMY09/11 were the most effective in detecting multiple infections (P<0.001). With each of the primer sets, a board range of HPV types could be identified, though there were several differences for a few genotypes. ConclusionThe substantial agreement between PCR-sequencing and HC2 for the detection of high-risk HPV (kappa=0.761) indicated that PCR-sequencing is also suitable for routine HPV screening.

Melanom Dışı Deri Kanserleri Gelişiminde Human Papillomavirüsün Rolü

Objective: Non-melanoma skin cancers (NMSC) represent the most frequent cancers among the Caucasian population worldwide. The possibility of the involvement of human pa - pillomavirus (HPV) in the development of these cancers is suggested by several studies. The pur - pose of this study is to evaluate the presence of various types of HPV in NMSC lesions of immunocompetent patients. Material and Methods: Biopsies taken from NMSC and non-sun ex - posed healthy skin from each of 32 immunocompetent patients were analysed for presence of HPV DNA by a polymerase chain reaction (PCR) using degenerate primers which can detect a broad spectrum of HPV types. Results: HPV DNA was detected in one of 21 basal cell carcino - mas, from one of two actinic keratosis and from none of 8 squamous cell carcinomas, and none of the control biopsy specimens. It was thought that in addition to factors associated with Turk - ish population, the specimens which were found to be HPV negative did not harbor HPV DNA, or although tissues harbor HPV DNA, it was not detected with the technique we used because of low viral load. Conclusion: We could not show an association between HPV infection and NMSC by using real-time PCR technique. Our data suggest that the use of MY09/MY11 primer pairs alone in PCR technique is not sufficient to evaluate the HPV prevalence in NMSC. We thought that studies with more patients and using more sensitive detection techniques which will be developed in the future are needed to show a significant association between HPV and NMSC

Comprehensive analysis of human papillomavirus prevalence and the potential role of low-risk types in verrucous carcinoma

The role of human papillomavirus (HPV) infections in the development of verrucous carcinoma, a well-differentiated variant of squamous cell carcinoma with difficult differential diagnosis, is controversial in the literature. In this study, we analysed verrucous carcinoma from different origins for the presence and activity of a broad spectrum of HPV types, and carefully reviewed the histopathological features. A random series of 27 formalin-fixed, paraffin-embedded specimens of verrucous carcinoma was taken, representing the head and neck region (n=6), anogenital area (n=16) and extragenital skin region (n=5). After review of the histological slides, all samples were subjected to different polymerase chain reaction-based HPV detection techniques, together detecting a total of 83 HPV types, including both mucosal and cutaneous types. Histological revision was carefully performed. Lesions with keratinised papillae, blunt stromal invaginations and minimal cytological atypia were considered verrucous carcinoma. Condylomatous lesions with viral changes were defined as giant condyloma. Verrucous lesions that did not meet those criteria were classified as verrucous hyperplasia. Tumours with stromal infiltration were considered as invasive squamous cell carcinoma. Histological revision revealed that 13 out of 27 cases were verrucous carcinoma (one showing a double infection with HPV 35 and 45), 5 invasive squamous cell carcinomas, 5 verrucous hyperplasia (one with a double infection with HPV 4 and 8), 1 pseudoepitheliomatous hyperplasia and 3 giant condylomas. All three giant condylomas were low-risk HPV positive (HPV 6 and 11) and showed active mRNA transcription. None of the HPV-positive samples tested positive for diffuse p16INK4A staining. In conclusion, our results do not support a causal role of HPV in the development of verrucous carcinoma. Testing for LR-HPV, particularly HPV 6 and 11, may help in the differential diagnosis of lesions suspicious of verrucous carcinoma as those testing positive for LR-HPV most likely represent giant condylomas.

High-risk and multiple human papillomavirus infections among married women in Can Tho, Viet Nam

The two currently licensed human papillomavirus (HPV) vaccines are highly efficacious in preventing cervical pre-cancers related to HPV 6, 11, 16 and 18. Before implementing a large-scale HPV vaccine campaign in Viet Nam, information about the prevalence of infection with the HPV vaccine types is required. This study was done in Can Tho, the province with the highest prevalence of cervical cancer in the south of Viet Nam, to explore the distribution of other high-risk types of HPV among married women in this province. The study employed a cross-sectional design with multistage sampling. A total of 1000 participants were randomly selected, interviewed and given gynaecological examinations. HPV infection status and HPV genotyping test were completed for all participants. A broad spectrum of HPV types was reported in this study. The prevalence of cases infected with HPV 16 and/or 18 was 7%; the prevalence of cases infected with other high-risk HPV types was 6%. The highest prevalence for single and multiple infections, as well as for high-risk infections, was reported for the youngest age group (less than 30 years). While it is relevant to implement an HPV vaccine campaign in Viet Nam due to the high prevalence of infection with HPV 16 and/or 18, it is important to note that one can be infected with multiple types of HPV. Vaccination does not protect against all types of high-risk HPV. Future vaccine campaigns should openly disclose this information to women receiving vaccines.

Comparison of HPV sign Genotyping Test with INNO-LiPA HPV Genotyping Extra assay on histologic and cytologic cervical specimens

An accurate tool for viral typing is important for management of patients with human papillomavirus (HPV) infection and to monitor HPV vaccine efficacy. This study evaluated the performance of the HPV sign® Genotyping Test by analyzing 87 archival cervical specimens and compared results with historical data by INNO-LiPA HPV Genotyping Extra assay. There was a substantial concordance for HPV detection in clinical samples (k 0.66), with an overall agreement rate of 85.1%. The genotyping overall agreement, considering one by one the HPV infection detected, was 95.7%. The HPV sign test showed, however, lower sensitivity than INNO-LiPA for HPV 31, 53, and 66. On the other hand, The HPV16 sensitivity was higher for HPV sign (90.0%, confidence interval [CI] 0.79–1.01) than for INNO-LiPA (83.3%, CI 0.70–0.97). Furthermore, HPV sign allowed identifying the presence of HPV16 intratype variants. In conclusion, HPV sign is a promising method for HPV genotyping and has revealed advantages in detecting a broad spectrum of HPV types and variants.

Human papillomavirus genotyping by 454 next generation sequencing technology

Background An accurate tool for human papillomavirus (HPV) typing is important both for management of patients with HPV infection and for surveillance studies. Objectives Design and evaluation of an HPV typing method based on 454 next generation sequencing (NGS) technology. Study design Development of an HPV typing method based on 454 NGS of HPV L1 amplicons generated with MY09/11-based primers. Evaluation of the NGS method in control samples and in a panel of cervical cytological samples. Comparison of the NGS typing method with cycle sequencing and with the reverse hybridization-based INNO-LiPA HPV Genotyping Extra assay (LiPA). Results In control samples carrying mixtures of HPV16 and HPV18 DNA, the NGS method could reliably detect genotype sequences occurring at a frequency of 1% in multiple infections with a sensitivity of 100 genome equivalents/μL. In cervical cytology samples, comparison with cycle sequencing demonstrated accuracy of HPV typing by NGS. The NGS method had however lower sensitivity for some HPV types than LiPA, conceivably due to the poor sensitivity of the MY09/11-based primers. At variance, LiPA could not detect HPV types which were present in low proportion in multiple infections (<10% of HPV reads obtained by NGS). In addition, NGS allowed identifying the presence of different variants of the same HPV type in a specimen. Conclusions NGS is a promising method for HPV typing because of its high sensitivity in multiple infection and its potential ability to detect a broad spectrum of HPV types, subtypes, and variants.

Abstract 4824: Little evidence of human papillomavirus in lung tumor tissue

Abstract Background: Lung cancer kills over one million people worldwide each year. Human papillomavirus (HPV) has been proposed as a potential cause. Over 50 studies have evaluated HPV in lung tumor tissue over the last 20 years with mixed findings, but most were severely underpowered or had other methodological limitations. While several HPV-associated cancers have been identified (e.g., cervical, tonsillar), the role of HPV in lung carcinogenesis remains controversial. Methods: We tested 450 lung cancer cases in the Environment And Genetics in Lung cancer Etiology (EAGLE) Italian population-based, case control study for HPV DNA. DNA was extracted from ethanol or formalin-fixed, paraffin-embedded tumor tissue under strict PCR clean conditions and was evaluated for HPV16 and 18 by type-specific, real-time PCR targeted to the E6 or E7 gene. The number of cell equivalents evaluable in these reactions was estimated by real-time PCR targeted to the human ERV3 gene. DNA purity was assessed by standard spectrophotometry (260/280 ratio). Results: 79.6% (n=358) of the 450 cases were male, 48.9% (n=220) were current smokers, 44.0% (n=198) were former smokers, 6.7% (n=30) were never smokers, and 0.4% (n=2) had unknown smoking status. 399 of 450 cases (88.67%) met quality indices for DNA quantity (at least 100 evaluable cell equivalents) and quality (260/280 ratio of 1.5 to 2.2). Of these, 398 (99.75%, 95% confidence interval: 98.61%-99.99%) were negative for HPV16 and 18. One case, a female former smoker with adenocarcinoma, was positive for HPV16 at low copy number (0.4 per 1000 cell equivalents). We are currently evaluating a subset of cases (n=100) for a broad spectrum of HPV types by PCR with SPF10 primers and a cocktail of conservative probes recognizing at least 54 mucosal HPV genotypes in a microtiter plate format (DEIA). To date no HPV has been detected in the 30 cases tested. Conclusions: Using extensive laboratory efforts to avoid DNA contamination and sensitive HPV DNA detection assays, we found no evidence to support a role for HPV in lung cancer in a representative Western population. Our results provide the strongest evidence to date to rule out a role for HPV in lung carcinogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4824.

Human papillomavirus in early laryngeal carcinoma

To examine the role of HPV status in the etiology, prognosis, and treatment of head and neck squamous cell carcinoma in early larynx malignancies. Retrospective. Thirty-eight cases of T1 or carcinoma in situ (CIS) laryngeal lesions were examined for the presence of human papilloma virus (HPV) using an inclusive polymerase chain reaction (PCR)/hybridization technique capable of identifying 37 HPV subtypes. HPV DNA was detected in 6 (16%) of the 38 lesions, representing HPV types 16, 26, 31, 39, and 52, and p16 tumor suppressor protein expression was confirmed in 10 representative cases. This HPV prevalence is higher than that noted in many previous laryngeal cancer studies, possibly due to the relatively large panel of subtypes screened for in this study. Identification of HPV-26, which has been associated with uterine cervical cancer, in an early laryngeal cancer specimen represents the first evidence of this subtype in a laryngeal carcinoma. Consistent with reports focusing on head and neck squamous cell carcinoma (HNSCC) arising from other subsites within the upper aerodigestive tract, patients with HPV-positive laryngeal carcinomas were of younger age and were somewhat less likely to have a history of tobacco use, although the latter of the two findings did not reach statistical significance. Our findings emphasize the presence of a broad spectrum of HPV types in a relevant proportion of early laryngeal cancers, and together with evidence of an association of HPV tumor status with a more favorable clinical course, provide a rationale for the routine HPV testing of small larynx lesions.

Human Papillomavirus-16 Is the Predominant Type Etiologically Involved in Penile Squamous Cell Carcinoma

Human papillomavirus (HPV) infections are suggested to be involved in the development of penile squamous cell carcinoma (SCC), but comprehensive studies to define the association are limited. Therefore, we performed molecular and serologic analyses for a broad spectrum of HPV types on a large series of 83 penile SCCs, and we compared serological findings to those of age-matched male controls (N = 83). Penile SCCs were subjected to detection and typing assays for mucosal and cutaneous HPVs and to subsequent, type-specific viral load and viral gene expression assays. Sera of patients and of controls were analyzed for type-specific mucosal and cutaneous HPV L1, E6, and/or E7 antibodies using bead-based, multiplex serology. HPV DNA of mucosal and/or cutaneous types was found in 46 of 83 (55%) penile SCCs. HPV16 was the predominant type, appearing in 24 (52%) of 46 of penile SCCs. The majority of HPV16 DNA-positive SCCs (18 of 24; 75%) demonstrated E6 transcriptional activity and a high viral load. Additionally, HPV16 molecular findings were strongly associated with HPV16 L1-, E6-, and E7-antibody seropositivity. Furthermore, serologic case-control analyses demonstrated that, in addition to the association of HPV16 with penile SCC, seropositivity against any HPV type was significantly more common in patients compared with in controls. HPV18 and HPV6 seropositivity were associated with HPV16-negative SCCs but were not correlated to molecular findings. HPV16 is the main HPV type etiologically involved in the development of penile SCC. Although individuals who develop penile SCC show a greater prior exposure to a broad spectrum of HPV types, insufficient evidence was found to claim a role for HPV types other than HPV16 in penile carcinogenesis.

Human papillomavirus testing methods.

Testing for human papillomavirus (HPV) relies exclusively on techniques of molecular biology using nucleic acid probes. Tests for HPV using nucleic acid probes have been commercially available since the late 1980s, but early tests were cumbersome, involving the use of nucleic acid probes labeled with radioactive phosphorus (32P). These early HPV tests did not achieve widespread use because they did not detect all oncogenic HPV genotypes. The current commercial HPV detection kit, Digene's Hybrid Capture 2 kit, detects virtually all high-risk oncogenic HPV types, as well as most low-risk nononcogenic HPV genotypes. The Hybrid Capture 2 test format is a proprietary nucleic acid hybridization signal amplification system owned by Digene Corporation. Virtually all test formats for DNA sequence analysis are amenable to applications intended to detect and perhaps quantify the various HPV genotypes. These methods can involve direct hybridization with complementary DNA probes, such as Southern blotting or in situ hybridization, signal amplification, such as the Hybrid Capture 2 method or target nucleic acid amplification, most notably the polymerase chain reaction (PCR). Polymerase chain reaction has been used for HPV detection, genotyping, and viral load determination. General or consensus primer-mediated PCR assays have enabled screening for a broad spectrum of HPV types in clinical specimens using a single PCR reaction. Following amplification using consensus primers, individual HPV genotypes are identified using a variety of methods. Using consensus primers in a test format known as real-time quantitative PCR (RQ-PCR), it is possible to generate viral load (concentration) data from reaction curves generated by monitoring PCR reaction kinetics in real time.

A broad spectrum of human papillomavirus types is present in the skin of Australian patients with non-melanoma skin cancers and solar keratosis.

Human papillomavirus (HPV) may play a role in the pathogenesis of non-melanoma skin cancer (NMSC) in epidermodysplasia verruciformis (EV) patients, but in the general population no specific HPV types have been associated with these lesions. Objectives To examine the spectrum of HPV types present in the skin and tumours of Australian patients with NMSC or solar keratosis (SK). Biopsies from tumours, and cotton swab samples of perilesional skin and buttock skin from each of 59 Australian patients with basal cell carcinoma (BCC), squamous cell carcinoma (SCC) or SK were tested for HPV DNA by polymerase chain reaction (PCR) using HPV consensus (FAP) primers and by type-specific primers for HPV 38 and candidate HPV 92. The identification of HPV type from consensus PCR was performed by sequencing and comparison with GenBank. In total, 49 of 59 (83%) patients harboured HPV DNA, which was detected in 28 of 64 (44%) biopsies, 48 of 64 (75%; P < 0.001) perilesional swabs and 36 of 59 (61%; P = 0.04) buttock swabs. Forty-five different HPV types/putative types were detected: 15 were previously characterized HPV types, 17 were earlier described putative types and 13 were new putative types. In addition, six subtypes and four variants of HPV sequences were identified. HPV types within the B1 group (EV HPV types) were found in 26 of 64 (40%) lesions, 44 of 64 (69%) perilesional swabs and 35 of 59 (59%) buttock swabs. HPV 38 was detected in 23 of 59 (39%) patients, and was found in seven of 16 (43%) SKs, but was less common in SCCs [three of 23 (13%); P = 0.037] and BCCs [four of 25 (16%); P = 0.056]. Candidate HPV 92 was found in seven of 59 (12%) patients. A broad spectrum of HPV types, the majority from the B1 group, was found in skin of Australian patients with skin tumours. HPV 38 was found significantly more often in SK than in SCC. However, the role of cutaneous HPV infection in the pathogenesis of NMSC remains elusive.

Improved detection of cutaneous human papillomavirus DNA by single tube nested ‘hanging droplet’ PCR

A single tube nested ‘hanging droplet’ PCR was developed for detection of cutaneous human papillomavirus (HPV) DNA of the phylogenetic group B1. The nested PCR was compared with a single round PCR method by testing 56 fresh biopsies from Australian skin tumour patients. HPV DNA was detected in 64% (36/56) of the biopsies by nested PCR and in 30% (17/56) by single round PCR ( P<0.001). HPV DNA was more often detected by nested PCR than by single round PCR in basal cell carcinoma [62% (16/26) vs. 19%; (5/26); P=0.003], squamous cell carcinoma [43% (7/16) vs. 25% (4/16)] and in solar keratosis [93% (13/14) vs. 57% (8/14); P=0.038]. The nested PCR and the single round PCR system detected 26 and 11 different HPV types/putative types/subtypes, respectively. Multiple types were found in eight samples by the nested PCR and two samples by single round PCR. The nested HPV PCR is more sensitive and capable of amplifying a broad spectrum of HPV types from skin tumours, but further improvements are needed before all HPV infections in skin can be detected by a single assay.

Papillomavirus in esophageal papillomas and carcinomas

Human papillomavirus (HPV) has been implicated as a possible etiological factor in the development of squamous-cell carcinoma of the esophagus. Published data demonstrating HPV DNA in these lesions have been conflicting, varying between failure to detect HPV DNA to detection of up to 60% to 70% of the biopsies harboring HPV DNA, mainly HPV 16 and HPV 18. We have analyzed esophageal carcinoma samples from 2 high-risk areas, China and South Africa, using a degenerate PCR approach. All amplified products were cloned and sequenced. A broad spectrum of HPV types was demonstrated in 10/29 samples from China and 9/34 samples from South Africa. HPV types detected included mucosal types HPV 6, 18, 51, 52 and 57; cutaneous types HPV 9, 20, 24 and 25; and the putative new HPV types DL231, DL428 and DL436, with HPV 6/51, 6/57, 20/9 and 20/DL231 occurring as double infections. HPV 6 predominated (4 samples) in 11 esophageal papillomas originating from patients in Europe and tested for HPV DNA. Other HPV types present included HPV 20, DL284 and DL436. Another putative new HPV type, DL416, was identified in a dysplastic lesion of the esophagus. Int. J. Cancer 80:681–684, 1999. © 1999 Wiley-Liss, Inc.

Papillomavirus in esophageal papillomas and carcinomas.

Human papillomavirus (HPV) has been implicated as a possible etiological factor in the development of squamous-cell carcinoma of the esophagus. Published data demonstrating HPV DNA in these lesions have been conflicting, varying between failure to detect HPV DNA to detection of up to 60% to 70% of the biopsies harboring HPV DNA, mainly HPV 16 and HPV 18. We have analyzed esophageal carcinoma samples from 2 high-risk areas, China and South Africa, using a degenerate PCR approach. All amplified products were cloned and sequenced. A broad spectrum of HPV types was demonstrated in 10/29 samples from China and 9/34 samples from South Africa. HPV types detected included mucosal types HPV 6, 18, 51, 52 and 57; cutaneous types HPV 9, 20, 24 and 25; and the putative new HPV types DL231, DL428 and DL436, with HPV 6/51, 6/57, 20/9 and 20/DL231 occurring as double infections. HPV 6 predominated (4 samples) in 11 esophageal papillomas originating from patients in Europe and tested for HPV DNA. Other HPV types present included HPV 20, DL284 and DL436. Another putative new HPV type, DL416, was identified in a dysplastic lesion of the esophagus.

Prevalence of human papillomavirus infection in premalignant and malignant lesions of the oral cavity in U.K. subjects: a novel method of detection.

To evaluate the possible role of human papillomavirus (HPV) in oral neoplasms, 28 oral squamous cell carcinomas (SCC) and 12 potentially malignant lesions were analysed for the presence of HPV DNA. A nested polymerase chain reaction (PCR) approach, using two sets of HPV consensus primers to the L1 region, was used, which was able to detect a broad spectrum of HPV types. HPV DNA was detected in 14/28 (50%) carcinomas and 4/12 (33%) precancerous lesions. A novel approach based on labelling the PCR products with 32P and the separation of radioactively labelled products on an 8% polyacrylamide gel increased the sensitivity of the detection and enabled the identification of the HPV types. The typing of HPV was subsequently confirmed by direct DNA sequencing. HPV 6 and HPV 16 were the only HPV types detected and seven tumours harboured both types. Our results suggest that HPVs may be an important aetiological factor in the development of oral cancer. The detection procedure ensured sensitivity and consistency of the detection of low copy numbers of the virus DNA. The presence of HPV in 33% of premalignant tissues suggests that HPV infection may be an early event in the malignant transformation of oral SCC. There was no statistically significant association between viral infection and tumour grade or stage.

Detection of human papillomavirus in squamous cell carcinomas of the lung by polymerase chain reaction.

Existing evidence supports the hypothesis that human papillomavirus (HPV) may play an etiologic role in the malignant transformation of squamous epithelial cells. Although HPV DNA has been identified in a high proportion of squamous cell carcinomas (SCC) of the cervix, anorectum, skin, and upper airways, few studies have tested for HPV in SCC of the lung. To confirm the presence of HPV in lung SCC, we tested for HPV DNA extracted from formalin-fixed tissues of 34 patients by polymerase chain reaction (PCR). DNA amplification was performed using HPV L1 consensus sequence primers (MY11 and MY09; Perkin-Elmer Cetus, Norwalk, CT) which recognize a broad spectrum of HPV types including 6, 11, 16, 18, 31, and 33, among many other known types, as well as at least 20 other unidentified types. PCR products were analyzed by agarose gel electrophoresis and Southern blot hybridization with [32P]-labeled generic HPV probes. HPV DNA positive cases were subsequently analyzed by slot-blot hybridization of the PCR products with specific probes for HPV types 6, 11, 16, 18, and 33. HPV type 18 was detected in two cases, including one case from a 44-year-old female and one from a 64-year-old male, with the remaining 32 cases negative. In situ hybridization for HPV DNA failed to detect HPV types 6/11, 16/18, or 31/33/35 in any of the cases. We conclude that a small proportion of cases of primary pulmonary SCC test positive for HPV type 18 but that the great majority of cases are not associated with HPV.

Age-dependence of human papillomavirus DNA presence in oral squamous cell carcinomas.

The aetiology of oral cancer is thought to be multifactorial. Apart from the two known major risk factors (tobacco and alcohol), a viral aetiology has been proposed, with special reference to human papillomavirus (HPV). 35 cases of oral squamous cell carcinoma (OSCC), seen at the Departments of Oral & Maxillofacial Surgery and Oral Pathology and Otolaryngology of the Free University of Amsterdam, were analysed as well as 12 biopsies of clinically and histologically normal gingival mucosa collected from healthy individuals after tooth extractions, using the polymerase chain reaction (PCR) and two different sets of primers that are able to detect a broad spectrum of HPV types. An overall HPV positivity of 54.3% in OSCC was found, the majority of positive cases (78.9%) harbouring HPV type 16. In contrast, no positivity for HPV was detected in the clinically normal oral mucosal samples analysed. Furthermore, a significant association between HPV presence and age was found: patients older than 60 years showed a lower prevalence of the virus (29.4%) compared with patients below this age (77.8%) (P < 0.05). The results from the present study suggest an association between HPV and OSCC, particularly in patients under the seventh decade.

Evaluation of Seroreactivity to Human Papillomavirus Type 16 Virus-like Particles in an Incident Case-Control Study of Cervical Neoplasia

An ELISA to detect serum IgG antibody response to human papillomavirus (HPV) type 16 virus-like particles (VLPs) was evaluated in a case-control study of cervical neoplasia, nested within a prospective cohort study. Subjects included 688 controls with continued normal cytology and 152 cases with confirmed incident squamous intraepithelial lesions who were tested for DNA for a broad spectrum of HPV types at cohort and follow-up of controls, 16.6% were seropositive compared with 30.8% and 52.4% of cases with low- and high-grade lesions, respectively. Of HPV-16 DNA-negative subjects, 16.5% were seropositive. Seropositivity increased from 22.2% in subjects who were HPV-16 DNA-positive by polymerase chain reaction once only (enrollment or follow-up) to 83.3% in those who were HPV-16 DNA-positive at both time points. These data imply that serum antibody to HPV-16 VLPs is a relatively sensitive indicator of persisting cervical HPV-16 infection.