Detection of human papillomavirus type 6f genome in nasal papillomatosis.

Five cases of nasal papillomatosis were studied clinicopathologically and virologically. In a case of recurrent papillomatosis of non-inverted type located on the nasal septum, human papillomavirus (HPV) DNA was detected by dot blot hybridization with an RNA cocktail probe of mucosal HPVs. In Southern blot hybridization, the DNA hybridized with that of HPV types 6 and 11 but not with those of types 16 and 18. Its restriction endonuclease-cleavage patterns corresponded well to those of HPV type 6f. These results suggested that HPV type 6 would also be associated with nasal non-inverted papillomatosis.     

Prevalence of human papilloma virus types and their influence on the course of the disease in children suffering from recurrent respiratory papillomatosis

The type of human papilloma virus (HPV) was determined in 26 children aged between 1 year 10 months to 15 years 5 months suffering from recurrent respiratory papillomatosis (RRP). Polymerase chain reaction identified DNA of HPV type 6, 11, 16 and 18. HPV DNA was detected in all the patients including fifteen patients infected with HPV type 11; seven patients infected with HPV type 6; four children infected with HPV type 6 and 11. Types 16 and 18 of HPV were not detected. The analysis of RRP course has found that laryngeal papillomatosis runs a more aggressive course in cases with HPV type 11 infection than in those with HPV type 6.     

喉乳头状瘤临床行为与细胞增殖活性关系的研究

利用ISH方法以及免疫病理手段对不同型的LP组织的标本进行PCNA、P53检测，探讨不同型HPV与LP的发生发展的相关机制，以寻求有效的检测方法帮助临床对LP的预后进行评估。标本选自1994年1月～1995年12月间我院收治的LP共36例。ISH法HPV6b／11阳性率为75％，明显高于HPV16和／或HPV18的表达。10例喉鳞癌各有1例HPV16、18阳性，无HPV6b／11阳性。ABC法行P53检测，36例LP标本中仅1例恶变组织阳性表达Ⅱ级（2．8％）；10例喉鳞癌中9例阳性表达（90．0％），其中Ⅱ级以上阳性表达6例（60．0％）。PCNA阳性表达27／36例（75％）；其中JOP组与AOP组阳性率有显著差异（P＜0．05）。本研究表明LP与HPV感染有极为密切的关系，认为HPV分型的检测在判断LP转归中有意义。PCNA阳性表达程度在预测LP肿瘤的活跃程度方面是一个很有意义的指标。P53蛋白表达在喉鳞癌与LP中有显著差异。     

鼻腔及鼻窦内翻性乳头状瘤与人乳头状瘤病毒感染及病毒亚型的关系

目的 采用多聚酶链反应(polymerase chain reaction，PCR)检测鼻腔及鼻窦内翻性乳头状瘤(nasal inverted papilloma，NIP)中人乳头状瘤病毒(human papilloma virus，HPV)DNA及其亚型，旨在探讨HPV在NIP发病中的作用及其与预后的关系。方法将28例鼻腔及鼻窦NIP分为无复发组、复发组、癌变组，用聚合酶链反应检测其HPV通用型、6、11、16及18型的感染情况，同时以10例慢性炎症鼻黏膜或炎症性鼻息肉病例为对照。结果28例NIP患者HPV-DNA总阳性率为75％(21例)。无复发组病例HPV-DNA阳性率为42％(5／12)，均为单一的低危型HPV感染(4例为HPV6型，1例为HPV11型)；复发组的13例和癌变组的3例中，均可检出HPV-DNA。复发者以检出HPV6、HPV11型DNA为主，4例具有双重感染。癌变者以检出HPV16、HPV18型DNA为主，其中2例为双重感染。结论NIP的发生与HPV感染密切相关。应用PCR检测HPV及各亚型感染情况有助于预测肿瘤的临床行为及预后，指导临床对高危患者进行重点监测。     

Detection of human papillomavirus genome in nasolaryngeal papillomas using digoxigenin labeled DNA probes

It is being reported that human papillomavirus (HPV) has been implicated in the pathogenesis of various neoplastic lesions of the genital organs. To investigate the etiological role of HPV and its types in nasolaryngeal papillomas, we retrospectively analyzed HPV genomes by nucleic acid hybridization methods; for detecting DNA and mRNA, we employed the recently developed nonradioactive (digoxigenin labeled) DNA probes and compared the results by radioisotope methods. In total, 43 cases of papillomatous lesions were examined. They were verruca vulgaris of the nasal vestibule (Nr = 2), nasal inverted papilloma (IP, Nr = 26), and laryngeal papilloma (Nr = 15). HPV types examined were type 2, 6, 11, 16 and 18. Two cases of verruca vulgaris were shown to contain HPV-2 DNA and its mRNA by in situ hybridization. HPV-11 DNA was detected in 3 cases (12%) of nasal inverted papilloma whereas HPV-16 was detected in 1 case (4%); the latter case was associated with squamous cell carcinoma. These results suggest that HPV may be implicated in the development of IP, and HPV-16 may play an important role in the malignant transformation of IP. In the cases of multiple laryngeal papilloma (Nr = 8, one juvenile type and 7 adult type), either HPV-6 or HPV-11 was detected at the high rate (6/8, 75%). The presence of the HPV genomes provides strong evidence for the HPV etiology of these laryngeal papillomas. Whereas in the cases of adult single laryngeal papilloma (Nr = 7), HPV was not detected. Technically, the sensitivity of digoxigenin (DIG) labeled DNA probe was almost same as 35S labeled probe by dot blot hybridization, thus we applied DIG labeled probe to Southern blot hybridization with low background. By in situ hybridization using digoxigenin labeled probes, the rates of HPV detection were almost equal to those by 35S labeled probes.(ABSTRACT TRUNCATED AT 250 WORDS)     

HPV DNA typing of adult-onset respiratory papillomatosis

Respiratory papillomatosis is a recalcitrant, human papillomavirus-induced disorder of the upper airways. Using in situ hybridization and biotinylated deoxyribonucleic acid (DNA) probes, the type of human papillomavirus (HPV) in nine adults was determined both at presentation and during subsequent recurrences. Fifty-three biopsies obtained from seven men and two women over a 10-year period were examined for HPV 6, 11, 16 and 18. All patients had either type 6 or 11. One dual infection was found. Despite intensive therapy with lasers with or without alpha interferon, no elimination or change in viral type occurred. Despite identical histologies, the amount of HPV DNA present in a given patient's papillomata varied markedly. The results suggest that adult-onset respiratory papillomatosis is due to HPV 6 and/or HPV 11, and that its recurrent nature, despite therapy, is due to persistence of a viral reservoir.     

Identification of HPV DNA in patients with juvenile-onset recurrent respiratory papillomatosis using SYBR Green real-time PCR

OBJECTIVE: Recurrent respiratory papillomatosis (RRP) is the most common benign neoplasm affecting the larynx and upper respiratory tract in children. Human papillomavirus (HPV) has been implicated as the cause of RRP, most commonly types 6 and 11. The present study was undertaken to evaluate the occurrence of HPV types in a group of patients with juvenile-onset RRP (JORRP). METHODS: The study group consists of 23 patients with JORRP. The clinical records of the patients were reviewed, and JORRP was classified as non-aggressive or aggressive. The laryngeal biopsies were taken and investigated for HPV DNA presence using real-time polymerase chain reaction (PCR) with a set of consensus primers (MY09/11). Viral typing was subsequently performed by real-time PCR with type-specific primers for HPV types 6, 11, 16, 18, 31, and 33. RESULTS: HPV presence was detected in all samples with amplifiable DNA. HPV-11 was revealed in 61.9% of the patients and HPV-6 in 23.8%. Double positivity for HPV types 6 and 11 was identified in 14.3%. Our findings suggest that RRP runs a more aggressive clinical course when HPV-11 infection is present (p=0.0265). CONCLUSIONS: Our results suggest a high frequency of HPV infection in the upper respiratory tract of the studied patients. We believe that the routine application of molecular techniques such as PCR for detection and analysis of HPVs in patients with RRP has diagnostic and prognostic significance.     

Is human papillomavirus involved in laryngeal neuroendocrine carcinoma?

The purpose of this study was to detect human papillomavirus (HPV) infection in laryngeal neuroendocrine carcinoma (LNEC) and to explore the possible relationship between HPV-induced malignant transformation and prognosis in LNEC. Ten cases of LNEC from a tertiary referral hospital were retrospectively analyzed. Clinical data were subtracted from patients’ files. Pretreatment biopsy material was tested for the presence of HPV6, 11, 16, and 18 using a PCR-based detection method. Immunohistochemical staining was performed for Ki-67, p16^INK4A, and p53 expression. All cases were negative for the low-risk HPV types HPV6 and HPV11 that are associated with laryngeal papillomatosis. High-risk HPV was detected in two cases; an atypical carcinoid was positive for HPV16 and a large-cell neuroendocrine carcinoma for HPV18. Both HPV-positive tumors had a high Ki-67 labeling index. Two of the four cases with a good response to therapy were hrHPV-positive (both HPV DNA positive) compared with none of the five poor responders. Our findings show that HPV may play a role in the pathogenesis of LNEC. The relationship between HPV, improved prognosis and good response to therapy for squamous cell carcinoma of the head and neck may also be true for a subset of LNEC.     

Presence of human papillomavirus type-6-related sequences in inverted nasal papillomas

Summary Twenty DNA samples obtained from seven cases of inverted papillomas, eight cases of nasal polyps and five cases of chronic sinusitis were investigated by Southern blot hybridization for the possible presence of sequences homologous to human papillomavirus (HPV) types 6, 11, 16 and 18. HPV type-6-related DNA was identified in one of the seven inverted papillomas. The restriction endonuclease cleavage patterns showed that this latter DNA is a new subtype of HPV type 6 DNA. In the other six papillomas and in all cases of nasal polyps and chronic sinusitis, no HPV sequence could be demonstrated, even under low stringent conditions (T _m–40°C). These results indicate that HPV infection might be one of the possible causative factors in the pathogenesis of inverted papillomas but is not essential for the induction of the tumor.     

Human papillomavirus identified by nucleic acid hybridization in concomitant nasal and genital papillomas.

Presence of human papillomavirus (HPV) as the etiologic agent in nearly all upper respiratory tract recurrent papillomas is well-established. The technique of nucleic acid hybridization now allows specific typing of HPV with a high degree of accuracy. This article reports a series of nine consecutive patients treated for nasal papillomas over the past 9 years. Eight of these patients had a personal history of genital papillomas (seven patients) or exposure (one patient). With the use of in situ hybridization and autoradiographic technique on paraffin-embedded tissue sections, HPV RNA type 6/11 was expressed in eight of nine nasal papillomas, and corresponding HPV types were also found in the two cases with which concurrent anogenital papilloma tissue was also available for analysis. Human papillomavirus RNA types 16 and 18 were not detected in any of the specimens. Signals of HPV messenger RNA type 6/11 were stronger in the fungiform areas than in the inverted areas of papillomas.     

Human papillomavirus type 11 DNA in squamous cell carcinomas and pre-existing multiple laryngeal papillomas

The case histories of 4 patients are presented, who developed an invasive squamous cell carcinoma from pre-existing multiple laryngeal papillomatosis (two juvenile-onset and two adult-onset) during a long latency period. A series of 14 routinely processed, paraffin-embedded biopsies obtained from these patients (including both benign papillomas and carcinomas) were analysed using in situ DNA-hybridization technique with 35S-labelled Human papillomavirus (HPV) DNA probes of HPV types, 6, 11, 16, 18 and 30. The lesions were also assessed by indirect immunoperoxidase (IP-PAP) technique to demonstrate the expression of HPV structural proteins. On light microscopy, morphology was consistent with HPV infection in all 14 biopsies. HPV antigen expression could not be demonstrated in any of the papillomas or carcinomas, however. HPV 11 DNA was present in high copy numbers in both the benign and malignant lesions of 2 patients, both presenting with the juvenile-onset disease. The present findings provide support for the role of HPV as an etiological agent in laryngeal squamous cell carcinoma, most probably acting synergistically with chemical or physical carcinogens (one patient received irradiation and 2 others were smokers). It seems clear that an infection by the 'low risk' HPV types 6 and 11 by no means excludes the possibility of developing a laryngeal malignancy, so far ascribed to the 'high risk' type HPV 16 only.     

Mucosal swabs detect HPV in laryngeal papillomatosis patients but not family members

Seven patients, aged 2-7 years, with active recurrent respiratory papillomatosis (RRP) attending the University of Michigan Pediatric Otolaryngology Clinic were studied to determine if human papillomavirus (HPV) is harbored in sites of the upper aerodigestive tract other than in the laryngeal papilloma itself. We also determined if close family members had detectable virus in their oral cavities. Noninvasive swabs of buccal mucosa, posterior pharynx, nasal vestibule, and tonsillar pillar of patients, as well as buccal mucosa and posterior pharyngeal swabs of family members were studied. Swabs of the patients' papillomas served as the positive controls. HPV was detected using polymerase chain reaction (PCR) amplification and Southern hybridization techniques. Six of seven patients had detectable HPV in papilloma and endolaryngeal swabs. Four were HPV type 6, and two were HPV type 11. The patient whose swab was negative for HPV was found to be biopsy negative for papilloma 3 weeks after a single laser excision which was performed 6 months prior to the endolaryngeal swab. HPV types 16, 18 and 31 were not found in any of the patients. No swabs from other sites in patients or family members were HPV positive despite the presence of adequate DNA in the swabbed material for successful amplification of beta-actin sequences. The absence of HPV (other than in the papilloma itself) in the upper aerodigestive tract of patients and caregivers is consistent with the absence of reported cases of horizontal transmission to siblings or other family members. The findings are also consistent with the conventional view that juvenile respiratory HPV is transmitted vertically from vaginal condylomas in the mother.     

Integration of human papilloma virus type 26 in laryngeal cancer of a child

Squamous cell carcinoma (SCC) in larynx is rare with children and adolescents. Usually larynx cancer is common with male smokers in the 7th decade. Among patients with no history of tobacco and/or alcohol consumption several factors have can play a role in the outbreak of laryngeal cancer: such as individual predisposition, radiation, gastroesophageal reflux, viral infection, dietary factors and environmental influences. In literature only few cases of laryngeal cancer with children are reported. Recent studies show that the most frequent laryngeal malignancy is the embryonal rhabdomyosarcoma. Besides the recurrent respiratory papillomatosis (RRP) based on an infection with human papilloma virus (HPV) types 6 and 11 (low risk) and types 16 and 18 (high risk) is known for a possible malignant transformation towards a SCC. HPV type 26 is only reported as low risk type HPV associated with cervical cancer. Final diagnosis often takes a long time. Initial symptoms such as hoarseness, cough or shortness of breath are often referred to more typical pediatric diseases or laryngeal development.     

Analysis of human papillomavirus in schneiderian papillomas as compared to chronic sinusitis and normal nasal mucosa.

Schneiderian papillomas (SPs) are tumours arising from the surface epithelium (schneiderian epithelium) of the nasal cavity and paranasal sinuses. Evidence points toward a viral etiology, specifically human papillomavirus (HPV). Although substantial data indicate HPV as a likely etiology, little is known about the role of HPV in benign nasal pathologies or in normal nasal mucosa. The objective of this study was to characterize the relationship between HPV and SP, chronic sinusitis (CS), and normal nasal mucosa. A case-control study was undertaken, matching patients with SP to patients with CS. Patients with normal nasal mucosa served as a control group. All patients had their tissues analyzed for the presence of various HPV subtypes using line blot assay. A total of 168 patients were identified (74 SP, 74 CS, 20 control). Of these, 70 (41.7%) had detectable deoxyribonucleic acid and 9 of 70 (12.9%) had detectable HPV of subtypes 6, 11, and 16. None had detectable HPV type 18. Significant differences were detected in the presence of HPV in the CS, SP, and control groups, as well as in the presence of low- versus high-risk subtypes among investigation and control groups. Significant differences exist in HPV infectivity among SP, benign nasal pathologies such as CS, and normal nasal mucosa. Human papillomavirus plays an important role, at least in part, in the development of SP, with types 6, 11, and 16 being more pivotal than other types. Line blot assay is a useful technique in identifying HPV in SP.     

HPV types causing juvenile recurrent laryngeal papillomatosis in South Africa

Objectives

To identify human papillomavirus (HPV) types associated with juvenile onset recurrent laryngeal papillomatosis (RLP) in southern Africa, to determine if there is a correlation between HPV type and disease aggressiveness and to determine the diagnostic and prognostic value of rapid molecular techniques for detection and typing of HPV using laryngeal biopsies.

Methods

Laryngeal biopsies from patients undergoing surgery for RLP were screened for HPV using conventional and real-time PCR techniques. Amplicons were sequenced to determine the HPV type involved. Clinical features were correlated with HPV type.

Results

HPV was identified in papillomata from 18 out of 19 patients. Only HPV-6 and HPV-11 were identified, with no co-infections. There was 100% concordance between conventional and real-time PCR techniques. Patients with HPV-11 disease required more procedures and tended to have higher Derkay scores than those with HPV-6 disease. The HPV types identified in our patients were genetically similar to HPV types from geographically distinct regions.

Conclusions

RLP in our patient population appears to be exclusively due to HPV-6 or HPV-11. HPV-11 disease appears to be more aggressive than HPV-6 disease. Identification of the HPV types provides motivation for inclusion of vaccines against these types in vaccination programs to protect women against infection and subsequently reduce the incidence of RLP.     

Human papillomavirus in sinonasal papillomas and squamous cell carcinoma.

The diagnostic and prognostic relevance of human papillomavirus (HPV) types 6, 11, 16, and 18 in squamous papilloma, inverted papilloma, and squamous carcinoma of the sinonasal epithelium was examined using the polymerase chain reaction (PCR) technique. Four (15%) of 26 squamous papillomas, 7 (24%) of 29 inverted papillomas, and 1 (4%) of 24 squamous carcinomas were positive for HPV when examined using the PCR amplification technique. Human papillomavirus 6 was present in 5 specimens (3 squamous and 2 inverted papillomas); HPV-11 was present in 6 specimens (1 squamous and 5 inverted papillomas); and HPV-18 was present in 1 of 24 squamous carcinomas. HPV-16 was not identified in any specimen. The proportion of tissue samples showing HPV presence, and the association of HPV types 6 and 11 with benign lesions and HPV-18 with malignant lesions, are both in accord with findings from prior investigations. Two major questions regarding nasal papilloma are the probability for lesion recurrence after surgical excision and the risk for malignant transformation. There is no unanimity of opinion regarding the prognostic value of histopathologic dysplasia to forecast these outcomes. HPV is etiologically related to a subset of sinonasal papillomas and squamous carcinoma, and those with benign and malignant clinical course are separable on basis of HPV type. Because of the paucity of these nasal lesions, a multi-institutional prospective collaborative study is the ideal way to address these questions.     

头颈部鳞状细胞癌中人乳头状瘤病毒DNA的表达

为探讨头颈部鳞癌与人乳头状瘤病毒（HPV）感染的关系和HPV在头颈癌中基因组型的分布与表达,应用聚合酶链反应技术制备非放射性探针标记物-地高辛标记HPV共有引物探针,对103树头颈部鳞癌（喉癌68例,鼻腔和上颌窦癌21例,鼻咽癌14例）和15例正常喉组织新鲜组织标本,进行HPV6,11,16,18,31,33,35,42,58共9型HPVDNA感染的检测;阳性者用多重引物PCR方法分型。喉癌HPV感染阳性率45.6％（31／68）,鼻腔和上颌窦癌阳性率为38．1％（8／21）,鼻咽癌阳性率14．3％（2／14）,15例癌周正常喉组织为HPVDNA阴性。HPVDNA型别分布在头颈部鳞癌中以HPV16、18型为主。头颈部鳞癌发生与HPV感染有相关性,喉癌发生与HPV感染尤为相关。     

Laryngeal papillomatosis: clinical, histopathologic and molecular studies

The clinical course and pathology of 57 patients with laryngeal papillomatosis were reviewed. Tissues from 26 patients were analyzed for human papillomavirus (HPV) DNA by Southern blot hybridization. Histopathologic evaluation of the papillomas showed no correlation with age of onset or clinical pattern of remission and recurrence. The pathology was characterized by abnormal squamous maturation with parakeratosis, retardation of superficial cell maturation, papillomatosis, and basal hyperplasia. HPV DNA was present in all lesions, with 92% containing either HPV-6 or 11. Latent HPV DNA was detected in clinically uninvolved tissues of 11 of 14 (78.5%) patients studied. There was no correlation between HPV type, histopathology and/or clinical pattern. Despite homogeneity of pathology, the clinical expression of laryngeal HPV infection varied widely. A mechanism for the pathogenesis of laryngeal papillomatosis, based on the concept of maturational arrest, is proposed.     

Tracing Human Papillomavirus DNA in Nasal Polyps by Polymerase Chain Reaction

Human papillomavirus (HPV) infections are related to the genesis of various benign and malignant human neoplasias. The HPV types 16 and 18 seem to be causally related to the development of most squamous cell carcinoma of the anogenital tract and a proportion of carcinomas of the upper aerodigestive tract. The near 100% positivity of the HPV types 6 and 11 in laryngeal papillomatosis is well established. We investigated whether HPV also plays a role in non-neoplastic mucosal entities such as sinunasal polyposis, the genesis of which has been discussed as being triggered by viral infections. On DNA from 39 sinunasal polyps (33 patients), polymerase chain reaction (PCR) was performed using