Detection of human papillomavirus infection in diseased and nondiseased sites of the respiratory tract in recurrent respiratory papillomatosis patients by DNA hybridization.

We examined 12 recurrent respiratory papillomatosis patients, who were undergoing treatment for recurrence of their disease, for the presence of human papillomavirus (HPV). Biopsies were obtained from their respiratory papillomas and nondiseased sites (NDS) of the respiratory tract: the nasopharynx, posterior tonsillar pillar, aryepiglottic fold, cervical trachea, intrathoracic trachea, and bronchi. The presence of HPV DNA was determined by using the ViraPap/ViraType DNA hybridization procedure. Two thirds of the patients were infected with HPV 6/11 in either the diseased papilloma or NDS: 50% of patients with papilloma specimens typed positive (6 of 12), and 40% of the patients typed HPV-positive in one or more biopsies from the NDS (4 of 10). No single NDS was more likely to be infected with HPV than any other. No oncogenic HPV types 16/18 or 31/33/35 were detected in the papillomas or NDS of these patients. Only patients with multiple, and not isolated, papilloma involvement were found to harbor HPV DNA in NDS; and 80% of those with infected NDS, compared to 20% of those without infected NDS, required a shorter (no more than 3 months) surgical treatment interval. Our results show that HPV infection frequently persists in adjacent, clinically normal sites, and suggest that the extent of NDS involvement may predict both the extent of disease and the likelihood of recurrence.     

Dissemination of squamous papilloma by surgical manipulation: A case report

The occurrence of multiple squamous cell papilloma in the upper and lower aerodigestive tract is well documented. While apparent inoculation to other areas in the lower respiratory tract at the time of bronchoscopy or tracheotomy is a frequently described occurrence, there have, however, been no reports of transmission of papilloma outside the respiratory tract as a result of surgical manipulation. The purpose of this paper is to report a case of squamous papilloma of the external auditory canal and tympanic membrane which resulted from apparent inoculation at the time of surgery for nasal cavity and nasopharyngeal papillomas together with myringotomies. This supports the hypothesis that an infectious agent, e.g., a virus could be the causative agent and that inoculation to other sites as a result of manipulation is possible. In light of our experience with this case we recommend specific guidelines for the handling of these patients and the protection of surgeons, anesthesia and operating room personnel. Hopefully this report will heighten awareness of papillomas as a potentially transmittable entity to sites outside the respiratory tract, and encourage others to approach these clinical situations with caution.     

Tonsil T cell immunity to human papillomavirus in the absence of detectable virus in healthy adults

BACKGROUND: Human papillomavirus (HPV) is known to infect the epithelium of the upper aerodigestive tract; however, major questions regarding prevalence and persistence of infection, and their relation to local immune response, remain unanswered. OBJECTIVES: To evaluate the tonsil T cell immune response to HPV and compare this to the frequency of detectable virus at this site. STUDY DESIGN: A cross-sectional study of cancer-free adults undergoing routine tonsillectomy. METHODS: Mucosal immune responses to recombinant HPV16 L2E6E7 and HPV6 L2E7 antigens were measured by tonsillar T-lymphocyte proliferation assay in 13 subjects. HPV deoxyribonucleic acid (DNA) was assessed by PCR and reverse line-blot hybridization in an expanded population of 44 subjects. RESULTS: Proliferative T-cell responses to HPV16 and HPV6 were identified in all patients. The presence of a CD45RO+ T cell population responsive to HPV6 L2E7 was confirmed in three of six subjects tested. There were no CD45RO+ responses to HPV16 L2E6E7 and no evidence of current or latent HPV infection of the tonsil. CONCLUSIONS: T cell memory to human papillomavirus can be identified in tonsil tissue from an adult population in the absence of concurrent HPV infection. How novel HPV vaccines might augment this preexisting cell-mediated immunity is an essential area for investigation.     

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 beta-globin primers for demonstration of amplifiable DNA in the tissue extracts. Consensus primers for the detection of several different HPV types were applied to the beta-globin-positive samples. The results were confirmed by Southern blot hybridization using consensus probes. Cycle sequencing was performed on the positive cases. All 39 samples showed positive signals for beta-globin. HPV-DNA investigations showed a slight positive signal in only 1 of the 39 investigated cases (2.6%). Further molecular investigations of this sample, including cycle sequencing, could not confirm this result. All the other tissue samples remained HPV-DNA-negative. Therefore, those HPV types readily detectable with the PCR primers and probes used are not frequently associated with sinunasal polyposis. The data confirm the hypothesis that HPV is correlated to a lesser extent to infectious mucosal lesions than to proliferative lesions. Furthermore, the results emphasize that the presence of HPV in specific lesions does not occur by chance, but represents a specific infection of the mucosa leading to proliferation and even to malignancy.     

Crohn's disease affecting the nasal mucosa

Crohn's disease is a chronic inflammatory disease of the gastrointestinal tract, associated with disorders of cell mediated and humoral immunity. The incidence of the disease is increasing, and it is commonest in young women. Any part of the gastrointestinal tract may be affected, and 9% of cases have oral lesions which are usually painful and coincide with periods of active intestinal disease. These oral lesions are areas of cobblestoned mucosa with deep ulcers and fissures; on histological examination there is perivascular inflammation with lymphocytic infiltration and in 10% non-caseating granulomas are seen. In the absence of any clinically detectable lesion, biopsies of buccal mucosa from patients with Crohn's disease show focal inflammation around salivary glands, and immunofluorescent studies have demonstrated antibodies to buccal mucosa in their serum. Involvement of the lower respiratory tract by Crohn's disease is rare, and although laryngeal involvement leading to upper airway obstruction has been described, involvement of the nose has not been previously reported.     

Ultraschallmikroskopie im oberen Aerodigestivtrakt

Purpose

Initial clinical experiences with ultrasound microscopy in the upper aerodigestive tract are described.

Materials and Methods

In the present pilot study, 20 healthy probands and 10 patients with lesions of the oral cavity, pharynx and larynx undergoing surgery were examined using a new prototype of an ultrasound microscope.

Results

A total of 24 normal and six pathological findings of the upper aerodigestive tract were assessed. These included normal mucosa of the floor of the mouth, the inner cheek, the palate and the vocal fold. In addition, invasive carcinoma of the floor of the mouth, the aryepiglottic fold and the vocal fold was found. Furthermore, a papilloma of the palatal arch and two epiglottic cysts were examined.

Conclusion

Our results demonstrate that ultrasound microscopy is also viable in hollow organs. Pathological lesions differed clearly from normal mucosa. However, to recognize the regular pattern of different lesions of the upper aerodigestive tract, further investigations need to be carried out with a larger number of patients.     

Human papillomavirus DNA in squamous cell carcinoma of the upper aerodigestive tract

Nineteen samples of DNA from squamous cell carcinoma of the upper aerodigestive tract were analyzed by Southern blot hybridization for the presence of human papillomavirus (HPV) DNA. Two samples of DNA contained HPV 16 DNA or its homologous sequence. In one maxillary carcinoma, the sequences homologous to HPV 16 were detected. In one tonsillar carcinoma, HPV 16 sequence was also shown to be present. The patients positive for HPV DNA were female and had neither smoking nor drinking habits. These results indicate that HPV infections may play a role in the development of some types of squamous cell carcinomas of the upper aerodigestive tract.     

Recurrent respiratory papillomatosis

Recurrent respiratory papillomatosis is a relatively rare disease caused by members of the human papilloma virus family. Lesions may occur anywhere throughout the respiratory tract but show a predilection for squamo-columnar epithelial junctions, frequently leading to hoarseness and upper airway obstruction. Rarely, it can progress to squamous cell carcinoma. The impact of recurrent respiratory papillomatosis on patients, their families, and the health care system is considerable. Unfortunately, despite extensive investigational studies, no cure is available for the disease. This article reviews the aetiology of and therapeutic options for recurrent respiratory papillomatosis.     

Human papillomavirus in laryngeal papillomas and in adjacent normal epithelium

Eleven adults with laryngeal papillomas were studied for the presence of human papillomavirus (HPV) DNA by in situ hybridization. As well as from the papillomas, three additional biopsies were taken from the normal-appearing mucosa as follows: the involved vocal cord, the opposite vocal cord (when the papilloma was unilateral), and from the ventricular fold on the side of the lesion. These normal tissues were analysed by polymerase chain reaction (PCR) to detect HPV DNA. All except one of the 11 papillomas contained HPV DNA; nine were HPV 6/11 DNA positive and one positive for HPV 16 DNA. The normal-appearing laryngeal mucosa harboured HPV DNA in eight out of 11 patients. The present results strongly support the concept that the adult-type laryngeal papilloma is an HPV-induced lesion, mostly due to HPV types 6 and 11. The persistence of HPV DNA in the adjacent normal epithelium is consistent with the frequent recurrence of these lesions.     

High incidence of malignant transformation of laryngeal papilloma in Taiwan

OBJECTIVES: Papillomas of the larynx include solitary laryngeal papilloma and recurrent respiratory papillomatosis. This study investigated the incidence of malignant transformation and assessed possible risk factors for laryngeal papillomas. STUDY DESIGN: A prospective, longitudinal study. METHODS: Twenty-six consecutive laryngeal papilloma patients were prospectively studied for 5 or more years, and each patient was periodically examined at 3 to 6 month intervals. A detailed epidemiologic questionnaire was administered at the initial visit. After enrollment, tissue obtained during each laryngeal surgery was examined by polymerase chain reaction assay for human papilloma virus (HPV) and typing. RESULTS: During 237 person-years of follow-up, six new, pathologically confirmed cases of laryngeal carcinoma were ascertained (incidence 2.5/100 person-years), and all were associated with HPV-6 or HPV-11. Malignant transformation revealed no correlation with the following: age less than 3 years at diagnosis, sex, history of tobacco use, history of alcohol consumption, family history of laryngeal cancer, or type of laryngeal papilloma. Laryngeal papilloma without demonstrable HPV DNA was the only significant risk factor for malignant transformation (P < .05). The cumulative risk of malignant transformation in subjects without demonstrable HPV DNA was significantly higher than that in HPV-positive patients (relative risk, 8.0; 95% confidence interval, 1.1-60.3; P = .05). CONCLUSIONS: A relatively high incidence of malignant transformation of laryngeal papilloma was noted in Taiwanese patients. Patients without demonstrable HPV DNA require more frequent follow-up and may benefit from anti-HPV vaccinations.     

Carcinoma ex-papilloma: Histologic and virologic studies in whole-organ sections of the larynx

A patient with adult-onset recurrent respiratory papillomatosis (RRP), initially diagnosed at age 28 years, was treated with radiation therapy due to the rapid regrowth of lesions. Following 6 years of apparently inhibited growth, papilloma recurred, and squamous carcinoma was diagnosed from a laryngeal biopsy. A spontaneous laryngocutaneous fistula developed, and laryngectomy was performed 14 years after irradiation. The laryngectomy specimen was snap frozen and representative tissues were stored frozen for viral studies. The larynx was whole-organ sectioned for histologic examinations; residual papilloma, as well as carcinoma, was observed. Koilocytosis and other virus-associated histologic changes were also found. HPV capsid antigen was present in papilloma, carcinoma, and clinically normal epithelium. HPV nucleic acids, conforming to HPV type 6, were present in keratin pearls and dysplastic cells. According to prior reports, carcinoma developing in preexisting papilloma arises from juvenile-onset RRP. Irradiated papilloma develop cancer at about 10 years, and the patients rarely survive. Nonirradiated cases develop cancer after 30 years, and some develop papilloma in the hypopharynx and trachea, but most patients survive. Irradiation is not an obligatory precursor for malignant transformation of cancer; however, until now there have been no case reports of favorable outcome after irradiation of papilloma.     

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.     

Papilloma viruses: group-specific antigens within lesions of the oral mucosa

The present study was undertaken with purpose to investigate the relationships between intraepithelial proliferations of malpighian mucosa and the presence of group specific papilloma virus antigens. The investigations allowing to give viral types or subtypes, the hybridization technics, are very heavy and not disponible in routine practice. The detection of widely distributed genus specific HPV antigen using PAP immunohistochemical labeling in different lesions and their association with displastic or neoplastic processes is presented. Only few studies were performed on oral mucosal proliferations, showing the association of these viral proteins with papillomatous or condylomatous lesions. Our work is directed towards the identification of the HPV antigens in more advanced lesions and in the mucosal epithelium surrounding and perhaps preexistent with the buccal cancer.     

p53-Mutationsnachweis in Abstrichen der Mundschleimhaut von Patienten mit oralem Plattenepithelkarzinom

Tumour-suppressor gene p53 encodes for an important cell-cycle regulatory protein and is therefore probably important for the development of many malignant diseases, e.g. squamous cell carcinoma of the mouth. This gene has mutated most frequently in connection with the development of cancer, so it has been well explored. Hence we chose it to find out whether swabs of the oral mucosa are suitable for supplying material for the detection of mutations in a gene that is connected with the development of oral squamous cell carcinomas, because swabs are easier to obtain than biopsies. We examined biopsies, swabs from the tumour, and swabs from mucosa that appeared healthy from 32 patients with oral squamous cell carcinoma and mucosal swabs from 35 healthy persons with polymerase chain reaction (PCR) and temperature gradient gel electrophoresis (TGGE). Fourteen of the 32 patients with a tumour showed mutations of p53, and in all cases the mutation could be demonstrated both in the biopsy and in the tumour swab. In four cases the mutation was also found in the swab of normal mucosa. Our investigations revealed that swabs are a suitable method for obtaining material for the detection of gene mutations in oral squamous cell carcinomas.     

Combined endolaryngeal videoendoscopic surgery and photodynamic treatment of patients with recurrent laryngeal and tracheal papillomatosis

Two-stage combined treatment of chronic recurrent papillomatosis of the larynx and trachea has been designed and tested in P.A. Herzen Research Cancer Institute. Stage I of the treatment consisted in endolaryngeal videoendoscopic surgery with Nd:YAG-laser destruction, argon-plasma coagulation and electroresection of the papilloma; stage II - postoperative photodynamic therapy (PDT) to prevent recurrence. In 1995-2007 the treatment was given to 32 patients aged 10-66 years with recurrent papillomatosis of the airways with the disease history 5-58 years. In two thirds of the patients papillomatosis involved several parts of the larynx and trachea. Squamous cell papilloma was accompanied with dysplasia of the first-second degree in 10 (31%) patients, dysplasia of the third degree - in 4 (12,5%), cancer in situ - in 3 (9,4%) patients. Human papilloma virus (HPV) was detected by hybridization in situ in 96%. The course of the treatment resulted in a complete regression (CR) of papilloma in 25 of 32 (78%), partial regression in 7 patients. The recurrence-free interval averaged 32 months (maximal 7 years) in 14 of 25 patients with CR. HPV was eradicated in a group of patients with persistent clinical remission. A 6 to 19 month follow-up recorded papilloma recurrence in 11 patients. The recurrence-free period increased 2,5-fold. In patients with dysplasia of degree I-III and cancer in situ (n=17) CR of dysplasia and preinvasive cancer foci was achieved in 15 (88%) patients.     

Acquired airway obstruction from histologically normal, abnormally mobile supraglottic soft tissues

In patients with nonmalignant upper aerodigestive tract abnormalities such as cysts, neoplasms, and laryngomalacia, airway obstruction has been described as a function of not only size and location of abnormal structures, but also of their mobility. This paper describes three patients who developed upper airway restriction with stridor due to abnormally redundant and mobile, but histologically normal, supraglottic mucosa. The specific mechanism of and structures involved in obstruction were clearly defined by videolaryngopharyngoscopy. Endoscopic microdissection and/or laser excision of the redundant supraglottic mucosa restored airway patency and completely relieved the obstructive respiratory symptoms. The pulsion effects of inspiratory airflow are postulated as the cause of these acquired lesions. Recommended treatment is reviewed.     

Pachyonychia congenita with laryngeal involvement

Pachyonychia congenita (Jadassohn-Lewandowsky Syndrome) is a rare autosomal dominant disorder characterized by nail dystrophy, hyperkeratosis of the palms and soles, leukoplakia of the mucosa of the upper respiratory tract and anus, follicular keratoses especially about the knees and elbows, and palmar and plantar hyperhidrosis. We present a patient with pachyonychia congenita and an exophytic lesion in the larynx at the posterior commissure. He is the youngest of 4 family members with this disorder covering 3 generations. Each of the 4 patients also exhibited both oral leukoplakia compatible with the Jadassohn-Lewandowsky syndrome (Ikonograph Dermatol. Lab., 1 (1906) p. 29), and subcutaneous cysts of the face and scalp as described by Jackson and Lawler (Ann. Eugenics (1951) 142.     

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.     

Alloplastic implants of the larynx

Biocompatible Teflon fluorocarbon polymer (Proplast) and porous polyethylene (Plasti-Pore) are porous alloplastic implant materials that are widely used in reconstructive head and neck surgery. These two materials were used in our study as replacement grafts for defects in three groups of 12 canines. For the first time, Plasti-Pore was found equal and perhaps superior to Proplast in deep implantation, this in the anterior cricoid cartilage when the internal and external perichondrium were preserved. Rejection of both substances occurred when the implant was exposed to air and aerodigestive tract contaminants after the removal of the perichondrium, subglottic, and upper tracheal mucosa. Airway exposure to the alloplasts, even when lined with buccal mucosa, resulted in infection and extrusion, but at a much slower rate.