Human papilloma virus in erosive oral lichen planus

Several types of human papilloma viruses (HPV) have been associated with benign and malignant squamous cell tumours of mucosal epithelium. To identify HPV in erosive oral lichen planus (OLPe), considered as a premalignant lesion, tissues from 20 patients were examined by Southern blot hybridization with 32P-labeled HPV DNA probes. Type 11 was found in 6 of the lesions while HPV types 6, 16 and 18 were not detected in any of the tissues examined. Using a type-specific polymerase chain reaction (PCR) assay for HPV-6, 11, 16 and 18, HPV-11 was detected in 8 of the samples (all of those positive by Southern blot), and, in addition, HPV-6 was found in 5 samples and HPV-16 in 3 samples. Overall, by the more sensitive PCR assay, 65% of samples were positive for HPV DNA. The finding of HPV DNA in many of the samples using two different techniques indicates a high prevalence of HPV in the OLPe afflicted oral mucosa. However, the role of HPV in the pathogenesis of OLPe has yet to be determined.     

Human papillomavirus and Epstein Barr virus in oral hairy leukoplakia among HIV positive Venezuelan patients

Oral hairy leukoplakia (OHL) is commonly found in individuals infected with HIV and represents the most frequent oral manifestation. The purpose of this study was to detect the presence of Human Papillomavirus (HPV) and Epstein Barr Virus (EBV) in OHL of HIV+ Venezuelan patients. We evaluated 21 HIV+ adult patients with clinically present OHL lesions: 11 under antiretroviral therapy, 10 without therapy, and 10 oral mucosal samples as controls. Nested-PCR was used to detect EBV and HPV infection. The INNO-LiPA HPV Genotyping v2 was applied to determine the HPV genotype. The EBV genome was found in 16/21 (76%) of the HIV+ patients with OHL. No difference was observed in EBV+ and EBV- patients related to antiretroviral therapy viral load and CD4+ Tcell coant. HPV-DNA was observed in 7/21 HIV positive cases (33%). The HPV genotypes detected were: 6, 11, 31, 33, 52, and 56/74. The most frequently HPV found was genotype 6 in 7/7, while two cases were HPV-11 and two HPV-52. Of the positive cases, 5/7 (71%) presented co-infection with more than one HPV genotype and 4/7 (57%) had HPV coinfection with high and low risk types. No case was EBV or HPV positive in the control group. In this study, a higher EBV prevalence was observed in OHL-HIV+ patients, confirming the etiologic role in this entity. A considerable number of cases were positive for HPV infection, and many patients presented coinfection with more than one HPV genotype as well as the presence of high oncogenic risk HPV in OHL.     

Human papillomavirus DNA types in squamous cell carcinomas of the head and neck.

Previous studies have found variable evidence suggestive of a role for human papillomavirus (HPV) in squamous cell carcinoma of the head and neck. In this study 49 cases of primary verrucous or squamous cell carcinoma from patients referred to a regional medical center were examined initially by Southern blot hybridization to detect HPV types 2, 6, 11, 13, 16, 18, and 32. Approximately 60% of carcinomas from certain head and neck sites, particularly the floor of the mouth, tongue, pharynx, piriform sinus, and larynx, were positive for episomal viral DNA of HPV-6, -11, -16, or -18. HPV DNA was found in some multiple tumors from separate sites of the same patient. Integration of viral DNA into the host cell chromosome was likely in a minority of the positive carcinomas, and no novel HPV DNA types were indicated by the hybridization analyses. Subsequently, DNA remaining from 30 of the carcinomas was examined by a more sensitive polymerase chain reaction amplification assay for DNA of HPV-6, -11, -16, and -18. Twenty-seven of the samples were positive for one or more HPV DNA types, with all positive carcinoma samples containing oncogenic HPV-16 or -18 DNAs. Almost all the patients examined were of the middle to older age group with a history of tobacco use. Although HPV infection of oral mucosa may be a frequent occurrence, a possible role for HPVs in the multifactorial etiology of head and neck carcinogenesis merits further epidemiologic investigation.     

Relations of human papillomavirus and lesions of the oral mucosa

Recently there has been an increased interest in the possible role that viruses and especially the Human Papilloma virus (HPV) could play in the etiology of lesions of the oral mucosa. A distinction has to be made between the so-called low-risk types of the virus (HPV-2, 6, 11, 13 and 32) which can be found in benign oral mucosal lesions, and the high-risk type (HPV-16), which predominantly is found in malignant oral mucosal lesions.     

人乳头状瘤病毒感染与儿童口腔黏膜良性上皮增生

目的 探讨儿童口腔黏膜良性上皮增生性病损与人乳头状瘤病毒(human papillomavirus，HPV)感染及其类型的关系。方法 选取四川大学华西口腔医学院病理科近10年30例儿童口腔黏膜良性上皮增生性病损的病例，复习其临床病理特征及切片，并采用免疫组织化学及原位杂交方法检测HPV共同抗原及其类型。结果 口腔鳞状细胞乳头状瘤(squamous cell papilloma，SCP)20例(66．7％)，尖锐湿疣(condyloma acuminatum，CA)6例(20．0％)，口腔黏膜局灶性上皮增生(focal epithelial hyperplasia，FEH)4例(13．3％)。HPV检测结果显示：HPV共同抗原阳性者占73．3％(22／30)，其中SCP占75．0％(15／20)，6例CA HPV共同抗原均为阳性，4例FEH中仅1例HPV共同抗原为阳性；HPV类型以高危型HPV16／18为主，占77．3％(17／22)，其次是HPV6和HPV11。结论 儿童口腔黏膜良性上皮增生性病损与HPV感染关系密切，病毒类型以高危型HPV16／18为主，其病毒类型是否与成人(以HPV6、11为主)不同尚待进一步研究。     

Detection of human papillomavirus DNA by in situ DNA hybridization and polymerase chain reaction in premalignant and malignant oral lesions.

The sensitivity of detection of human papillomavirus (HPV) DNA in premalignant and malignant oral lesions by in situ hybridization (ISH) and polymerase chain reaction (PCR) were compared. With both methods HPV DNA was found in 4 of 24 cases of epithelial dysplasia, 4 of 14 cases of verrucous hyperplasia, and 1 of 10 cases of squamous cell carcinoma. The 10 cases of smokeless tobacco keratoses and 3 cases of verrucous carcinoma that we examined were all negative for HPV DNA. The PCR for the E6 open reading frame of HPV-16 correctly identified all cases that were positive by ISH. Only a single case that was positive by PCR was negative by ISH for HPV DNA. However, the PCR demonstrated the presence of HPV-16 infection in one case, which had hybridized most intensely with the probe for types 31/33/35 in the ISH. This discrepancy probably is due to the high degree of cross-hybridization in the ISH assay. PCR appears to be an effective technique for identifying HPV-16 DNA sequences in biopsy material from premalignant and malignant oral lesions.     

Detection of human papillomaviruses of high oncogenic potential in oral squamous cell carcinoma in a Venezuelan population

OBJECTIVE: The aim of this work was to detect and typify human papillomaviruses (HPV) in oral squamous cell carcinoma (OSCC) in a Venezuelan population. MATERIAL(S) AND METHODS: Eighteen tissue samples were obtained from biopsies, formalin-fixed, and paraffin-embedded; 16 were diagnosed as SCC. We isolated DNA from paraffin-embedded tissue; two to three sections of 5 microm were obtained and resuspended in digestion buffer and proteinase K. Five microliters of the aqueous phase was used for polymerase chain reaction (PCR). The PCR for HPV amplification was carried out with consensus primers for L1 region (MY09 and MY11) and beta-globin gene was used as internal control. The viral types were determined by molecular hybridization with a mix of probes for high/intermediate and low HPV oncogenic risk types. RESULTS: The HPV-DNA was detected in 50% (eight of 16) of the SCC cases. Of these HPV-DNA-positive samples, 68% were histopathologically diagnosed as moderately differentiated SCC. The most common anatomical location was the alveolar ridge mucosa. All positive biopsies contained high oncogenic HPV types. CONCLUSIONS: We observed a high prevalence of HPV infection of high oncogenic potential types in patients with SCC in our studied group. The moderately differentiated SCCs were more associated to HPV infection. These differences could be influenced by nutritional, environmental and genetical factors in our population but further studies should be carried out to determine these aspects.     

Detection of human papillomavirus in head and neck tumors with DNA hybridization and immunohistochemical analysis.

The presence of human papillomavirus (HPV) DNA in oral, sinus, pharynx, and larynx lesions of Japanese patients was studied by Southern blot hybridization under less stringent (25% formamide, 42 degrees C) and stringent (50% formamide, 42 degrees C) conditions. Three samples from 10 benign tumors, and 3 of 30 malignant tumors, contained HPV DNA or HPV-related sequences. The HPV DNAs harbored in three laryngeal papillomas were HPV-11, -6, and -6 or -11, respectively. The HPV DNA and viral capsid antigens were easily detected by in situ hybridization, Western blotting, and peroxidase-antiperoxidase staining. However, neither the typical restriction pattern of HPV DNA nor viral antigen was identified in the malignant tumors, suggesting that subgenomic fragments remained integrated in the host cell DNA.     

High prevalence of human papillomaviruses in the normal oral cavity of adults

Human papillomavirus (HPV) infection in the normal oral cavity was studied by the sensitive polymerase chain reaction (PCR) using primers for the L1 region of human papillomavirus DNA and high fidelity amplification system. Cells were scraped from the oral mucosae of 7 (mean age; 42 years) and 30 (mean age; 32 years) volunteers with and without skin warts, respectively. Human papillomavirus DNA was detected in 30/37 (81.1%) specimens and their copy numbers per cell were 10(-1) to 10(-4) (mean, 10(-3)). The human papillomavirus types determined by PCR-based sequencing analysis were HPV-18 (26/30; 86.7%), -61 (18/30; 60%), -59 (7/30; 23.3%), -16 (2/30; 6.7%), -6 (1/30; 3.3%) and an unknown type (HPV-X71) (1/30; 3.3%). Multiple human papillomavirus types were present in 17/30 (56.7%) specimens. HPV-6 was detected in 2 of 7 skin warts and differed from the human papillomavirus types of the corresponding oral specimens. These data suggest that human papillomavirus infection in the oral mucosa occurs much more frequently than previously considered.     

艾滋病毒感染患者口腔黏膜疣状肿块人类乳头状瘤病毒感染的分型研究

目的探讨艾滋病毒感染患者口腔戮膜疵状肿块人类乳头状瘤病毒(1-IPV)感染及感染类型。方法应用 HPV Ll区通用引物Gp5十//G p6+和1-IPV特型引物(HPV6/II,16,18,31,33),通过聚合酶链反应(PCR),对34例艾滋病毒感染患者口腔豁膜沈状肿块人类乳头状瘤病毒(HPV) DNA进行检测。结果艾滋病毒感染患者口腔豁膜优状肿块中,HPV感染率为88.2%;亚型HPV6/II感染率为47.06%, HPV16感染率为II.76%, 1U〕V 18感染率为 2.94 % , HPV31感染率为5.88%。结论艾滋病毒感染患者口腔庆状肿块和H PV感染关系密切,而且大多数和低危型HPV6/II感染有关,少数病例和高危型HPV16,18,31感染有关;同一病例可以感染两种以上HPV亚型。     

The prevalence and distribution of human papillomavirus genotypes in oral epithelial hyperplasia: proposal of a concept

Background:  Evidence is accumulating for the aetiological role of human papillomavirus (HPV) in the pathogenesis of potentially malignant oral mucosal lesions and squamous cell carcinomas.
Methods:  Paraffin tissue sections from 49 patients with 'white patches' of the oral mucosa were investigated histologically, by broad-spectrum PCR followed by genotyping and chromogenic in situ hybridisation (CISH).
Results:  Histologically, 33 flat hyperplasias and 16 papillary hyperplasias were diagnosed. Twenty-two of 28 samples studied (78.6%) were positive for HPV DNA by PCR and six were negative. The following HPV types were detected in decreasing order of prevalence: HPV 35, HPV 6, HPV16, HPV 53, HPV 18, HPV 51 and HPV 55. Seventeen samples (60.7%) contained high-risk HPV DNA. Using CISH, ≥ 1 HPV signals were detected at least in a few epithelial cells in 95% of cases studied. All but one case were positive with the high-risk HPV probe and all HPV infections contained low viral load. Concordant positive results both by PCR and CISH were detected in 14 of 19 cases (73.7%) analysed.
Conclusions:  The high prevalence of HPV infection in hyperplastic 'white patches' of the oral mucosa supports the putative role of HPV at an early stage of oral carcinogenesis. These results further indicate that the majority of white oral mucosal lesions – flat, exophytic, wart-like or papillary proliferations – could be considered as the clinical manifestations of oral HPV infection. This finding has clinical relevance regarding therapy and patient management and may help in elucidating the role of HPV infection in oral carcinogenesis.     

In situ hybridization analysis of human papillomavirus DNA in oral mucosal lesions.

Commercial biotinylated DNA probes specific for human papillomavirus (HPV) types 6 and 11; 16 and 18; and 31, 33, and 35 were used for in situ hybridization analysis of 105 oral mucosal specimens from 5 cases of verruca vulgaris, 15 cases of condyloma acuminatum, 30 cases of squamous papilloma, 20 cases of hyperkeratosis/acanthosis, 15 cases of epithelial dysplasia, 5 cases of carcinoma in situ, and 15 cases of squamous cell carcinoma. Positive hybridization signals were found in 26 specimens (24.8%). Only HPV-6/11 was detected. HPV DNA occurred significantly more often (p less than 0.005, chi-square analysis) in condyloma acuminatum (100%) and verruca vulgaris (100%) than squamous papilloma (13.3%), hyperkeratotic/acanthotic lesions (10%), and malignant and premalignant lesions (0%). The tongue (19.1%) and labial epithelium (17.1%) were infected most frequently. Nuclear reaction products indicating HPV infection were associated primarily with koilocytes. These results demonstrate the usefulness of commercial biotinylated probes for HPV DNA analysis in routine paraffin-embedded lesion specimens. They confirm HPV involvement in benign lesions of the oral mucosa but fail to associate HPV infection with oral cancer and precancer.     

Concurrent HPV infection in oral and genital mucosa

Screening for human papillomavirus (HPV) types was performed by a PCR- based assay on 29 women (mean age 34.0 years, range 21-48 years). HPV-DNA was demonstrated in 16 women (55.2%), with a detection rate of 37.9% in the oral cavity and 34.5% in the genital tract. HPV-16 was the most prevalent genotype (53.8%), followed by HPV-6, which was present in 34.6% of the positive samples. Other types were more rarely detected. Five subjects showed concurrent genital tract and oral cavity infections but HPV type-specific concordance was detected in only 3 patients. Multiple HPV infections were found in 9 of the 26 positive samples, where HPV-6 appeared frequently associated with the other types. These data confirm the occurrence of mixed HPV infections and the wide diffusion of different types of HPV in the genital mucosa and in the oral cavity; they also stress the need to utilize diagnostic methods with a wide typing capacity.     

Human papillomavirus frequency in oral epithelial lesions

BACKGROUND: Oral human papillomavirus (HPV) prevalence varies according to geographical occurrence, the type of lesion, and the method of diagnosis. The polymerase chain reaction method (PCR) appears to be more sensitive and can be easily applicable to epidemiologic studies. OBJECTIVES: To determine the frequency of HPV and its genotypes in oral lesions among patients attending a reference clinic of a university hospital. METHODS: PCR was performed to identify HPV DNA from samples of oral epithelial lesions in 80 patients. For HPV DNA amplification, MY09/MY11 consensus primers were used and specific genotypes were identified through restriction fragment of length polymorphism (RFLP) pattern. RESULTS: HPV DNA was present in 11.3% of patients, and the identified genotypes were 6b, MM4 (W13B), and MM9 (PAP238A). CONCLUSIONS: HPV DNA frequency in patients with oral epithelial lesions was 11.3%. The genotypes MM4 and MM9 are uncommon in oral lesions, and they are characterized as high-risk HPV types in those types of lesions.     

HPV DNA in clinically different variants of oral leukoplakia and lichen planus

OBJECTIVES: Our objectives were to determine the prevalence of human papillomavirus (HPV) infection in oral leukoplakia (OL) and oral lichen planus (OLP) in comparison with that in healthy oral mucosa, also conditionally to age, gender, smoking, and drinking habits of patients, so as to investigate any possible association of HPV infection with a specific clinical variant of OL or OLP. STUDY DESIGN: We did research on HPV DNA in 68 cases of OL (homogeneous form [H] in 45 cases and nonhomogeneous form [non-H] in 23 cases), and in 71 cases of OLP (nonatrophic/erosive form [non-AE] in 27 cases, atrophic/erosive form [AE] in 44 cases). HPV DNA was investigated in exfoliated oral mucosa cells by nested PCR (nPCR: MY09-MY11/GP5-GP6) and the HPV genotype determined by direct DNA sequencing. RESULTS: HPV DNA was found in 17.6% of OL, in 19.7% of OLP, and in 5.6% of controls, with a statistically significant higher risk of HPV infection in both lesion groups (for OL: P=.01; Odds Ratio [OR]=3.64; 95% CI: 1.21-10.80; for OLP: P=.005; OR=4.17; 95% CI: 1.41-12.18). Demographic variables analysis showed that the only significant association was between HPV status and current smoking in OL patients (OR'=3.40; 95% CI: 1.0-11.59). HPV DNA was found in 20% of H OL and 13% of non-H OL, without any association with the clinical variant (P=.73; OR=0.60; 95% CI: 0.14-2.48). HPV DNA was found in 18.5% of non-AE OLP and 20.4% of AE OLP, without any significant association with the clinical variant (P=.84; OR=1.13; 95% CI: 0.335-3.816). HPV-18 was the most frequently detected genotype (9/12 and 10/14 of HPV-positive OL and OLP, respectively), followed by HPV-16 (2/12 OL and 2/14 OLP), HPV-33 (1/12 OL), HPV-31 (1/14 OLP), and HPV-6 (1/14 OLP). CONCLUSIONS: An increased risk of HPV infection was found in OL and OLP; however, no specific clinical variant of OL or OLP was noted to be associated with HPV infection. It is not possible to predict the likelihood of HPV infection from the clinical features of OL and OLP.     

Southern blot hybridization and PCR in detection of oral human papillomavirus (HPV) infections in women with genital HPV infections

The presence of human papillomavirus (HPV) in biopsies taken from clinically normal buccal mucosa (n = 212) and clinical lesions (n = 60) was examined by Southern blot hybridization (SBH) using 32P-labelled HPV DNA probes. Furthermore, one hundred formalin-fixed, paraffin-embedded biopsies were analyzed by using polymerase chain reaction (PCR), combined with dot blot hybridization and biotinylated HPV DNA probes. With SBH and PCR, 15.4% and 29.4% of the biopsies, respectively, contained HPV DNA. In clinically normal epithelium, 15.6% and 23.1% of the samples were HPV-positive with SBH and PCR, respectively. The HPV types detected in the genital and oral mucosa of index patients differed in all except two cases. Histology could not be relied on distinguishing HPV DNA positive and HPV DNA negative samples. Hand warts were encountered significantly more frequently in patients with a concomitant oral HPV infection. To conclude, oral HPV infections as detected by SBH and PCR are surprisingly common, but similar to the genital tract, the virus seems to exist in a latent form in the vast majority of cases. The frequent concomitant finding of skin warts and oral HPV infection may suggest some kind of HPV-specific immunosuppression.     

National prevalence of oral HPV infection and related risk factors in the U.S. adult population

This article reviews the rapidly growing evidence that oral human papilloma viruses (HPV) infection contributes to the risk of oral squamous cell carcinoma. It also reports the first nationally representative estimates of oral HPV prevalence in the United States adult population. An estimated 7.3% (95% CI: 6.0, 8.9) of the U.S. population had one or more oral HPV types detected in oral rinse; 3.1% (95%CI: 2.4, 3.9) of the U.S. population had one or more oncogenic HPV types. A substantial excess risk of HPV infection in men is not explained by education, smoking, age of sexual debut, or number of lifetime sex partners. Based on the published finding from a case-control study, where there was an odds ratio of 2.6 (95% CI: 1.5, 4.2) for the association of head and neck cancer oncogenic oral HPV infection, the estimated population attributable risk for head and neck cancer in the U.S. population was 4.7%. In other words, there would be a 4.7% reduction in incidence rate of head and neck cancer in the United States if oncogenic HPV infection could be prevented. The results also provide population data that help evaluate the likely public health benefits of prophylactic vaccination against oral HPV acquisition.     

Human papillomavirus (HPV) DNA in focal epithelial hyperplasia by in situ hybridization

Eighteen cases of focal epithelial hyperplasia (FEH) were investigated for the presence of human papillomavirus (HPV) group specific antigen by immunocytochemistry and HPV types 1, 6, 11, 13, 16, 18 and 32 by DNA in situ hybridization employing biotinylated probes. Seven (39%) specimens demonstrated the presence of HPV group specific antigen. Fifteen (83%) specimens were positive for HPV DNA: 9 (60%) showed HPV 32, of which 6 were on non-keratinized mucosa and 3 on border of keratinized and non-keratinized mucosa; 5 (33%) showed HPV 13, 4 lesions on keratinized mucosa and 1 on non-keratinized mucosa; 1 (7%) specimen on non-keratinized mucosa showed HPV-11 related. Two specimens on different sites from one patient showed the same HPV type and one patient had, in addition to FEH, a squamous papilloma also demonstrating the same HPV type. Results show a specific HPV distribution pattern in the epithelium indicating areas of high viral concentration adjacent to areas of low or no viral concentration. This study also indicates the possibility of tissue-site specificity or a latent infection and the possibility of a yet unidentified HPV type associated with FEH. It is suggested that future monitoring of patients be carried out with special reference to HPV type and anatomical distribution pattern for FEH lesions.     

In situ DNA hybridization analysis of oral papillomas, leukoplakias, and carcinomas for human papillomavirus.

Twenty-one papillomas, 23 ordinary benign keratoses, 13 smokeless tobacco keratoses, 10 verrucous hyperplasias, 10 verrucous carcinomas, 17 squamous cell carcinomas, 3 epithelial dysplasias, and 6 lichen planus lesions were evaluated for human papillomavirus (HPV) types 6/11, 16/18, and 31/33/35, with biotinylated double-stranded DNA probes by in situ hybridization. Sixty-two percent (13/21) of oral squamous papillomas were positive for HPV DNA. HPV DNA types 6 and 11 demonstrated the strongest reactivity. Of the 13 cases, 10 also showed some reactivity with HPV-16/18 and -31/33/35. None of the cases of keratoses, epithelial dysplasia, squamous cell carcinoma, verrucous hyperplasia, verrucous carcinoma, or lichen planus were positive for HPV DNA. This study confirms the consistent and frequent finding of HPV DNA in oral squamous cell papillomas and the inconsistency of being able to identify HPV DNA in keratotic, premalignant, or cancerous lesions of the oral mucous membranes.     

HPV in oral squamous cell carcinomas of a Brazilian population: amplification by PCR

Human Papilomaviruses (HPV) are a group of viruses associated with benign and malignant lesions of cutaneous and mucosal epithelia. Some "high risk" HPV types, especially HPV 16 and 18, are strongly correlated with cervical and anogenital cancers and are also related to the genesis of oral squamous cell carcinomas (OSCC). The aim of this work was to investigate the incidence of HPV infection in 40 paraffin-embedded or fresh specimens of OSCC, using PCR amplification of the viral DNA. Literature based primers (GP5+/GP6+) were used in order to amplify HPV DNA from the L1 gene, present in more than 22 types of HPV. A condyloma case with HPV 16 and 18 detected by in situ hybridization was used as a positive control. Amplification of HPV was observed only in the positive control. No squamous cell carcinoma cases showed DNA viral amplification. Absence of HPV DNA amplification by PCR in the analyzed specimens of OSCCs suggests that this virus not always plays a role in the carcinogenesis process. Discrepancy with some studies found in the literature may be related to methodology or population differences.