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.     

Focal epithelial hyperplasia arising after delivery of metal-ceramic fixed dental prosthesis

Focal epithelial hyperplasia (FEH) is a human papillomavirus (HPV)-induced alteration of the oral mucosa that presents with a clinically distinct appearance. While other HPV-infected lesions such as squamous papilloma, verruca vulgaris, and condyloma acuminatum involve the skin, oral mucosa, and genital mucosa, FEH occurs only in the oral mucosa. The affected oral mucosa exhibits multiple papules and nodules with each papule/nodule being flat-topped or sessile. The affected region resembles the normal color of oral mucosa rather than appearing as a white color since the epithelial surface is not hyperkeratinized. Almost all cases present with multiple sites of occurrence. This rare, benign epithelial proliferation is related to low-risk HPV, especially HPV-13 and -32, and is not transformed into carcinoma. We report a case of FEH that arose on the attached gingiva of an East Asian male adult related to prosthesis without detection of any HPV subtype in HPV DNA chip and sequencing.     

Papillary lesions of the oral cavity: relationship to human papillomaviruses

Human papillomaviruses are a group of genetically related organisms that infect stratified squamous epithelium. Unlike many other viruses that infect oral epithelium and induce lysis of the cells they penetrate, HPVs induce proliferative changes in these cells that result in both benign and malignant tumors. The common skin wart (verruca vulgaris) is induced by HPV 2 and 4. Genital warts (condylomas) and the common solitary oral papilloma are associated with HPV 6 and 11. Either HPV 13 or 32 causes focal epithelial hyperplasia. All of these wart-like lesions are benign growths of the stratified squamous lining of the oral cavity and lips and can be treated by surgical excision or laser ablation. HPV 16 and other less frequently encountered genotypes are associated with uterine cervix cancer in 95 percent to 98 percent of cases, and the evidence for a causal role is robust. There are emerging data that implicate HPV in certain subsets of oral cancer, particularly those that arise in the oropharynx/tonsillar region. Some instances of the various histologic subtypes subsumed under proliferative verrucous leukoplakia and verrucous carcinoma also harbor HPV.     

Oral squamous papillomas: detection of HPV DNA by in situ hybridization

Oral squamous papillomas were segregated from other papillary lesions on the basis of histopathologic features. Twenty representative papillomas were evaluated for the presence of papillomavirus genus-specific antigen with the use of an immunoperoxidase technique. These same tumors were analyzed for human papillomavirus (HPV) types 2, 4, 6, and 11 with biotinylated full-length double-stranded DNA probes by in situ hybridization. Only one case exhibited papillomavirus antigen reactivity. Alternatively, seven of twenty cases (35%) yielded positive results for HPV 6 or 11 DNA; one papilloma exhibited a dual infection with both HPV 2 and 6 when assayed under conditions of high-stringency hybridization. It is concluded that some oral squamous papillomas harbor HPV genotypes akin to those encountered in genital tract condylomas. Viral DNA can be detected in the absence of capsid antigen immunoreactivity, thereby obviating the use of antigen detection assays for determining the presence or absence of virus.     

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.     

Human papillomavirus (HPV) DNA sequences in oral precancerous lesions and squamous cell carcinoma demonstrated by in situ hybridization

A series of routinely processed, paraffin-embedded biopsies from 73 surgically treated oral precancerous lesions (OPL) (22 cases), and oral squamous cell carcinomas (SCC) (51 cases), was first screened using an in situ DNA hybridization technique with a human papillomavirus (HPV) DNA probe cocktail containing the 35S-labelled DNA of HPV types 6, 11, 13, 16, 18 and 30. The specific HPV types in lesions shown to contain HPV DNA in this procedure were further analysed by using in situ hybridization and the 6 HPV DNA probes separately. A total of 12/73 (16.4%) of the lesions proved to contain HPV DNA; 6/51 (11.8%) carcinomas and 6/21 (28.6%) dysplasias. The most frequent sites of HPV DNA-positive lesions were palate (4/7; 57%), followed by the floor of the mouth (2/8; 25%), the tongue and gingiva (11.8%). HPV 13 or HPV 30 were not found in any of the lesions studied. HPV 11 DNA was demonstrated in 2 mild dysplasia lesions, but not in carcinomas. One additional mild dysplasia proved to contain HPV 6 DNA. HPV 16 DNA was present in 5 biopsies; 3 carcinomas and 2 dysplasias. In one of the HPV 16-positive carcinomas, HPV 18 DNA was simultaneously present. HPV 18 alone was found in 3 additional carcinomas and in one moderate dysplasia lesion. The results confirm the recently reported evidence on HPV involvement in OPL and oral cancer. The implications of these findings are discussed in terms of the possible HPV etiology of oral SCC. The use of the in situ DNA hybridization as a powerful tool (enabling the localization of specific HPV DNA sequences and the proper classification of the lesion at the same site) in the study of routinely processed oral biopsies is strongly advocated.     

Human papillomavirus (HPV) DNA sequences in oral precancerous lesions and squamous cell carcinoma demonstrated by in situ hybridization

A series of routinely processed, paraffin-embedded biopsies from 73 surgically treated oral precancerous lesions (OPL) (22 cases), and oral squamous cell carcinomas (SCC) (51 cases), was first screened using an in situ DNA hybridization technique with a human papillomavirus (HPV) DNA probe cocktail containing the 35S-labelled DNA of HPV types 6, 11, 13, 16, 18 and 30. The specific HPV types in lesions shown to contain HPV DNA in this procedure were further analysed by using in situ hybridization and the 6 HPV DNA probes separately. A total of 12/73 (16.4%) of the lesions proved to contain HPV DNA; 6/51 (11.8%) carcinomas and 6/21 (28.6%) dysplasias. The most frequent sites of HPV DNA-positive lesions were palate (4/7; 57%), followed by the floor of the mouth (2/8; 25%), the tongue and gingiva (11.8%). HPV 13 or HPV 30 were not found in any of the lesions studied. HPV 11 DNA was demonstrated in 2 mild dysplasia lesions, but not in carcinomas. One additional mild dysplasia proved to contain HPV 6 DNA. HPV 16 DNA was present in 5 biopsies; 3 carcinomas and 2 dysplasias. In one of the HPV 16-positive carcinomas, HPV 18 DNA was simultaneously present. HPV 18 alone was found in 3 additional carcinomas and in one moderate dysplasia lesion. The results confirm the recently reported evidence on HPV involvement in OPL and oral cancer. The implications of these findings are discussed in terms of the possible HPV etiology of oral SCC.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Identification of human papillomavirus types in oral verruca vulgaris

Eleven oral verruca vulgaris specimens were examined for the presence of papillomavirus structural antigens by reaction with antibody to type-common antigens and detection by the avidin-biotin-peroxidase complex method. The specimens were also examined by in situ hybridization with biotin-labelled human papillomavirus (HPV) DNA to determine the specific HPV types present in the lesions. Six of the 11 specimens were positive for papillomavirus structural antigens. Of these 6, 5 hybridized to the HPV Type 2 (HPV2) probe and one to the HPV4 probe.     

Condyloma acuminatum and human papilloma virus infection in the oral mucosa of children

PURPOSE: The purpose of this study was to investigate the clinicopathological features of oral condylomas in children and condylomatous lesions of their mothers. Moreover, the authors sought to determine the mode of transmission of this disease and to find the genotype of human papilloma virus (HPV) in the children's oral condyloma. METHODS: Nine instances of oral condyloma acuminatum in children and lesions in their mothers were reviewed. Their HPV genotypes were evaluated by in situ hybridization (ISH). RESULTS: This study revealed that the lesions appeared during 3 years of age and the most common location was the hard and soft palate. Seven of the 9 mothers had experienced vulva and/or oral cavity condylomata during pregnancy. Social evaluation confirmed sexual abuse in 1 girl, and probable sexual abuse in another girl. The results of ISH demonstrated HPV 16/18 DNA being positive in 5 of the 9 cases, and HPV 6 and HPV 11, HPV 6 and HPV 16/18, HPV 6, and HPV 11 DNA being positive, respectively, in 1 case. HPV DNA types in mother-child pairs were not concordant. CONCLUSIONS: Oral condyloma acuminatum in children is probably induced by HPV 16/18. The mode of transmission by sexual abuse is the most likely route. Prenatal transmission of HPV to children is rare. This study provides further confirmation of possible different genotype and transmission in oral CA of children and adults.     

儿童口腔尖锐湿疣的临床表现及HPV检测

目的:探讨儿童口腔粘膜尖锐湿疣的病毒类型、传播途径、临床病理特点及预后等。方法:回顾6 例被确诊为口腔粘膜尖锐湿疣患儿的临床特点及HE 切片,并对其中5 例采用免疫组化染色及原位杂交检测人乳头状瘤病毒(HPV)DNA。结果:儿童口腔尖锐湿疣多发生在2 岁左右,发病部位多位于腭部,并且多数有家族感染史,镜下见棘层上部或角化层常出现灶性凹空细胞,免疫组化检测结果显示5 例HPV 共同抗原全部阳性,5 例中有4 例HPV16P18- E6 阳性;原位杂交结果显示仅1 例HPV6 和HPV11 同时阳性,另1 例初发时HPV6 阳性而复发后呈阴性。结论:儿童口腔尖锐湿疣的病毒类型、传播途径可能与成人不同。     

Identification of human papillomavirus types in oral verruca vulgaris

Abstract Eleven oral verruca vulgaris specimens were examined for the presence of papillomavirus structural antigens by reaction with antibody to type-common antigens and detection by the avidin-biotin-peroxidase complex method. The specimens were also examined by in situ hybridization with biotin-labelled human papillomavirus (HPV) DNA to determine the specific HPV types present in the lesions. Six of the 11 specimens were positive for papillomavirus structural antigens. Of these 6, 5 hybridized to the HPV Type 2 (HPV2) probe and one to the HPV4 probe.     

Detection of human papillomavirus in oral lesions using commercially developed typing kits

Biopsy material from 20 oral lesions (19 condylomas and 1 squamous papilloma) previously shown to contain human papillomavirus (HPV) 6 and HPV 11 sequences by in situ hybridization were examined using 3 commercially available HPV typing kits. Sensitivity and specificity were compared with in-house methods. Previous in situ hybridization had detected HPV 6b in 11 (55%) of the biopsies, HPV 6 and 11 in 7 (53%) and HPV 11 alone in 1 biopsy. Only one of the commercial assays (assay 1) detected HPV in all 20 biopsies (11 positive for HPV 6b only, 1 for HPV 11 only and 7 for HPV 6b and 11). The wide spectrum probe of assay 2 detected HPV in only 10 (50%) of the biopsies, and in a further 2 biopsies the hybridization results were difficult to interpret because of background staining. Assay 3 used a combined HPV 6/11 probe and detected HPV in 15 (75%) of the biopsies. Clear hybridization signals were demonstrated in the intermediate and upper layers only of squamous epithelium, as expected from the known association of HPV replication with epithelium differentiation. In most specimens background levels were not a problem, and all commercial assays were easy to use. The findings are discussed in the context of the digestion procedures, sensitivity of the probes provided and the conditions of hybridization, all of which would influence the detection of HPV     

Oral inverted ductal papilloma associated with condyloma acuminata and HPV in an HIV+ patient

Oral inverted ductal papillomas are rare benign tumours of minor salivary glands. A case that appeared in a middle-aged HIV+ woman simultaneous to two condyloma acuminata on the lips is described. The presence of human papilloma virus (HPV) 11 DNA was demonstrated in all the samples using polymerase chain reaction. HPV could play an important role in the aetiology and pathogenesis of these lesions in this patient.     

Oral wart associated with human papillomavirus type 2

More than 100 human papillomavirus (HPV) types have been identified to date. Of these, 24 types have been described as being associated with oral lesions. HPV-2 has been frequently associated with skin lesions, but the reports of oral lesions as features of mucosal infection are limited. A biopsy specimen of an oral wart on the right palate was taken from a 48-year-old man and examined for the presence of HPV The sections showed papillary growth of the epithelium with hyperkeratosis and parakeratosis, and koilocytotic changes of the cells located in the upper layers of the oral squamous cell epithelium. These histological features corresponded well to those of verruca vulgaris on the skin. Immunohistochemically, papillomavirus genus-specific capsid antigen was detected in most of the koilocytotic cells. In addition, Southern blot hybridization analysis revealed that the lesion harbored HPV-2 DNA. In situ hybridization with a biotinylated HPV-2 DNA probe clearly demonstrated viral DNA in the nuclei of squamous cells, which were located in a deeper layer of the epithelium than viral antigen-positive cells.     

The histologic differentiation of oral condyloma acuminatum from its mimics

OBJECTIVE: The purpose of this study was to determine which histologic features could enable one to distinguish oral condyloma, as defined by the detection of human papillomavirus (HPV) DNA through in situ hybridization, from its mimics. STUDY DESIGN: Thirty-two paraffin-embedded specimens from 28 patients that were clinically suggestive of oral condyloma were analyzed histologically and through in situ hybridization with a consensus HPV probe. RESULTS: HPV DNA was detected in 17/32 (53%) of the lesions; no additional positive cases were detected after polymerase chain reaction amplification. Only 5 of the 17 virus-positive cases were considered to be histopathologically unequivocal for condyloma. The histologic features significantly associated with HPV detection were nonuniform perinuclear halos, often in association with epithelial crevices (P =.02), and papillomatosis (P =.02). Each of the 17 patients who were HPV-positive had either HPV-6 or HPV-11, a finding that is similar to those for condyloma involving the penis and vulva/vagina. CONCLUSION: Differentiation between oral condyloma and its mimics is best accomplished by using a combination of histologic, clinical, and in situ viral studies.     

Human papillomavirus (HPV) types 2 and 57 in oral verrucae demonstrated by in situ hybridization

Twenty-one cases of verrucae vulgaris (oral warts) were investigated for human papillomavirus (HPV)-group specific antigen by immunocytochemistry and for HPV types 1, 2, 4, 6, 11, 16, 18 and 57 by DNA in situ hybridization with biotinylated probes. Twelve (57%) cases demonstrated the presence of HPV-group specific antigen. Fifteen (71%) cases showed the presence of HPV DNA, 13 of which (87%) demonstrated both HPV types 2 and 57 in the same cells and 2 of which (13%) demonstrated only HPV 2. Six cases were negative for HPV 2 and 57 and all 21 cases (100%) were negative for HPV types 1, 4, 6, 11, 16 and 18. Results indicate the association of a new and as yet unidentified HPV type, closely related to HPV 2 and 57, with oral warts. The identification of both cutaneous type HPV 2 and another type closely related to HPV 2 and 57 in oral verrucae on keratinized and non-keratinized mucosal surfaces indicates the possibility of a latent infection; three patients had a history of warts on their hands, suggesting autoinoculation. This study indicated that future investigations of oral warts, based on a correlation of clinical and histological features with HPV types by DNA in situ hybridization, are called for.     

Detection of human papillomavirus-related oral verruca vulgaris among Venezuelans

A sensitive in situ hybridization test under low stringency conditions (LCS) with a set of digoxigenin-labeled human papillomavirus mixed probes (D-L HPV MP) revelaed a positive reaction in 8 of 10 cases of oral verruca vulgaris (OVV), Ages ranged from 5 to 37 years with a mean of 14.5 years. 50% of all cases were located intraorally on the hard palate, followed in frequency by the commissures. These preliminary findings provide evidence of the role of HPV in OVV from a sample of the Venezuelan population. We show that in situ hybridization conducted under LSC is useful in HPV detection (regardless of the type) and the digoxigenin-labeling system is a rapid, relatively easy and specific method. In addition, this technique permits the retrospective evaluation of routinely processed material, thus widening the investigative spectrum for HPV.     

Non radioactive in situ hybridization for detection of human papilloma virus DNA in squamous cell carcinoma of tongue.

Previous studies indicate that HPV type 16 and 18 (HPV) are associated with squamous cell carcinoma of the head and neck. In this investigation we evaluate in our hospital 253 patients with oral squamous cell carcinoma of the tongue not related to tobacco or alcohol with a tumor index of T2 NO MO between 1981 and 1991. From 12 patients we were able to obtain tissue. For detection of human papilloma virus, DNA sequences 6, 11, 16, 18, 31 and 33 in paraffin-embedded human tissue biopsies a non-radioactive in situ hybridization procedure was utilized. Approximately 60% of the carcinoma of the tongue are positive for episomal viral DNA 6, 11, 16 and 18. These results confirms that HPV infection may play a possible role in the multifactorial etiology of carcinogenesis of squamous cell carcinoma of the tongue and probably acting synergistically with other carcinogenesis.     

口腔鳞状细胞乳头状瘤组织中HPV DNA的原位杂交研究

目的 探讨人乳头瘤病毒（ｈｕｍａｎ ｐａｐｉｌｌｏｍａｖｉｒｕｓ，ＨＰＶ）感染与口腔鳞状细胞乳头状瘤（ｓｑｕａｍｏｕｓ ｃｅｌｌ ｐａｐｉｌｌｏｍａ，ＳＣＰ）的发生之间的关系。方法 应用地高辛标记的ＨＰＶ６／１１和ＨＰＶ１６／１８核酸探针分别在３０例口腔ＳＣＰ组织上进行原位杂交，检测口腔ＳＣＰ组织中ＨＰＶ ＤＮＡ的特征。结果 ＨＰＶ６／１１ ＤＮＡ阳性１６例（５３％），ＨＰＶ１６／１８ＤＮＡ未检出，ＨＰＶ６／１１ＤＮＡ阳性细胞多数分布在鳞状上皮的表层、中层和基底层。结论 原位杂交方法可以检测口腔ＳＣＰ组织中ＨＰＶ ＤＮＡ的存在并能准确组织定位，进一步支持ＨＰＶ６／１感染与口腔ＳＣＰ的发生密切相关。