Xenograft model for therapeutic drug testing in recurrent respiratory papillomatosis.

OBJECTIVE: Identifying effective treatment for papillomatosis is limited by a lack of animal models, and there is currently no preclinical model for testing potential therapeutic agents. We hypothesized that xenografting of papilloma may facilitate in vivo drug testing to identify novel treatment options. METHODS: A biopsy of fresh tracheal papilloma was xenografted into a NOD-scid-IL2Rgamma(null) (NSG) mouse. RESULTS: The xenograft began growing after 5 weeks and was serially passaged over multiple generations. Each generation showed a consistent log-growth pattern, and in all xenografts, the presence of the human papillomavirus (HPV) genome was confirmed by polymerase chain reaction (PCR). Histopathologic analysis demonstrated that the squamous architecture of the original papilloma was maintained in each generation. In vivo drug testing with bevacizumab (5 mg/kg i.p. twice weekly for 3 weeks) showed a dramatic therapeutic response compared to saline control. CONCLUSION: We report here the first successful case of serial xenografting of a tracheal papilloma in vivo with a therapeutic response observed with drug testing. In severely immunocompromised mice, the HPV genome and squamous differentiation of the papilloma can be maintained for multiple generations. This is a feasible approach to identify therapeutic agents in the treatment of recurrent respiratory papillomatosis.

Tracheal ulcer due to Epstein-Barr virus-positive diffuse large B-cell lymphoma of the elderly.

A 74-year-old man was referred to our hospital because of a tracheal stenosis circumscribed with soft tissue density and a left pulmonary nodule. Open biopsy of a right submandibular lymph node revealed diffuse large B-cell lymphoma, and the malignant cells were positive for Epstein-Barr virus gene products. Bronchofiberscopy revealed a tracheal necrotizing ulcer. After chemotherapy, the tracheal ulcer resolved. To our knowledge, this is the first report of a case of Epstein-Barr virus-positive diffuse large B-cell lymphoma of the elderly with a tracheal ulcer.

Papillary squamous cell carcinoma of the trachea associated with human papillomavirus-18 infection.

A 75-year-old man presented with the chief complaints of coughing and dyspnea. Chest computed tomography revealed narrowing of the trachea. A tumor measuring 40×33×15 mm in size that caused -90% reduction in the cross-sectional area of the trachea 4 cm above the carina was cauterized with high-frequency electrocautery via a rigid bronchoscope. A pathological examination revealed papillary squamous cell carcinoma with immunohistochemical p16 overexpression. Multiplex polymerase chain reaction confirmed human papillomavirus type 18 DNA in the tumor. At the 12-month follow-up visit following the administration of radiotherapy, the patient exhibited no local recurrence or distant metastasis.

A case of multiple squamous cell papillomas of the trachea.

We report a case of 68-year-old woman suffering from breathlessness on exertion with stridor. A chest computed tomography showed a tumor arising from the posterior wall of the trachea. The diagnosis was squamous cell papilloma of the surgically removed tumor, which had caused the asphyxiation. After removal of the tumor, the patient received radical therapy: semiconductor laser transpiration. Polymerase chain reaction (PCR) detected human papilloma virus (HPV) type 6, thought to be the cause of the respiratory papilloma.

Epidemiology of recurrent respiratory papillomatosis.

Recurrent respiratory papillomatosis (RRP) was first described in the 1800s, but it was not until the 1980s when it was convincingly attributed to human papilloma virus (HPV). RRP is categorized into juvenile onset and adult onset depending on presentation before or after the age of 12 years, respectively. The prevalence of this disease is likely variable depending on the age of presentation, country and socioeconomic status of the population being studied, but is generally accepted to be between 1 and 4 per 100 000. Despite the low prevalence, the economic burden of RRP is high given the multiple procedures required by patients. Multiple studies have shown that the most likely route of transmission of HPV in RRP is from mother to child during labor. Exceptions to this may include patients with congenital RRP who have been exposed in utero and adult patients who may have been exposed during sexual contact. Although cesarean section may prevent the exposure of children to the HPV virus during childbirth, its effectiveness in preventing RRP is debatable and the procedure itself carries an increased risk of complications. The quadrivalent HPV vaccine holds the most promise for the prevention of RRP by eliminating the maternal reservoir for HPV.

Recurrent respiratory papillomatosis with lung involvement.

The case of a 27-year-old male with recurrent respiratory papillomatosis including lung involvement is presented. Laryngotracheal papillomatosis with lung involvement is a rare entity associated with human papillomavirus infection. Computed tomography (CT) was essential in guiding diagnostic and therapeutic approaches. Knowledge about the findings of this disease is needed for correct diagnosis, since findings are nonspecific. Lesions may show malignant transformation; regular follow-up with CT is essential.

An adult case of multiple squamous papillomas of the trachea associated with human papilloma virus type 6.

A 72-year-old woman with primary biliary cirrhosis complained of dry cough and wheezing. Chest computed tomography showed a tumor arising from the posterior wall of the trachea. Bronchoscopic examination revealed that the tumor was cauliflower-like, with two small polypoid tumors. They were diagnosed as multiple squamous papillomas. The main tumor was recurrent and removed by repeated microwave coagulation therapy (MCT) through bronchoscopy, whereas the two polypoid tumors were likely to disappear spontaneously. Human papilloma virus (HPV) type 6 DNA was detected in the tumor by polymerase chain reaction (PCR) amplification, suggesting that this virus was the cause of her papillomas.

Human papillomavirus genome detection by in situ hybridization in fine-needle aspirates of metastatic lesions from head and neck squamous cell carcinomas.

BACKGROUND: Patients with head and neck squamous cell carcinoma (HNSCC) often present with metastatic disease. The diagnosis of metastatic lesions usually is determined by fine-needle aspiration. Human papillomavirus (HPV) is now being considered as a causative agent in a subset of HNSCC. The objectives of this study were, first; to search for the presence of HPV DNA by in situ hybridization (ISH) in metastatic lesions from HNSCC using alcohol-fixed, archival, cytopathologic material; second, to characterize the cytologic features of HPV-positive metastatic lesions of HNSCC; and, third, to determine whether there is a correlation between the presence of HPV DNA and the origin of metastatic lesions. METHODS: The authors performed chromogenic ISH analysis for HPV DNA on fine-needle aspiration materials from metastatic lesions from 26 patients with HNSCC. Along with the ISH analysis, a detailed cytologic review was performed, and cytopathologic features were recorded. The HPV DNA status in metastatic lesion was correlated with cytopathologic features and primary tumor location. RESULTS: The integration of HPV DNA was visualized microscopically on tumor cell nuclei in 15% of aspirates. The anatomic locations of the study samples were as follows: 16 lymph node aspirates (11 cervical lymph nodes and 5 lymph nodes at other sites other), 5 tracheostomy sites, and 5 miscellaneous sites located on the head and neck area. Cytologic review revealed 13 keratinized and 13 nonkeratinized metastatic tumors. HPV DNA was detected in four metastatic sites (three lymph nodes and one tracheostomy site). All HPV DNA-positive tumors were of the nonkeratinizing type (P < 0.05; Fisher exact test). The origins of HPV-positive tumors included two laryngeal sites, one nasopharyngeal site, and one oral cavity site. CONCLUSIONS: The current findings showed that archival cytology slides can be used for HPV DNA detection with ISH. The results also showed that HPV DNA-containing HNSCC has distinctively nonkeratinizing cytologic features. The authors concluded that HPV DNA not only is involved in the initiation of tumoral processes but also plays an important role in the development of metastatic disease.

Human papillomavirus and squamous cell carcinoma in a solitary tracheal papilloma.

We report the case of a 52-year-old woman suffering from breathlessness on exertion. A chest radiograph identified a lesion in the intrathoracic trachea. A tumor 0.4 x 0.5 x 0.7 cm in size causing an approximately 80% reduction in the cross-sectional area of the trachea 3 cm above the carina was removed, and histology showed moderately differentiated squamous cell carcinoma. Intrathoracic resection of the tumor and anastomosis was performed through a right lateral thoracotomy. The final histology examination showed atypical papilloma. Polymerase chain reaction-restriction fragment length polymorphism confirmed human papillomavirus-6b in the tumor. The patient remained well 18 months after surgical intervention without recurrence.

Molecular transformation of recurrent respiratory papillomatosis: viral typing and p53 overexpression.

Recurrent respiratory papillomatosis (RRP) is a histologically benign disease of the larynx, trachea, and bronchi. Here we report on the histologic and molecular characteristics of 7 cases of malignant transformation of RRP to squamous cell carcinoma (SCCA). The clinical histories of 7 patients with RRP who developed SCCA were carefully reviewed. Sequential biopsies were available from 5 of the 7 cases of spontaneous transformation of RRP to SCCA and were reviewed. In addition, p53 protein overexpression and human papillomavirus (HPV) typing for all cases was examined. The average age of patients with juvenile-onset RRP was 3 years, and that of patients with adult-onset RRP was 31 years. The average age of onset of transformation to SCCA was 28 years. All patients had laryngeal involvement with RRP, and 3 of the 7 patients had tracheal extension of disease. Five patients were tracheotomy-dependent. Four of the 7 patients developed SCCA of the lung, while 3 patients developed laryngeal SCCA. There was no consistent histologic progression from squamous papilloma to papilloma with dysplasia, and all but 1 of the SCCAs were well differentiated. The overexpression of p53 protein was variable in each of the 5 patients. We detected HPV types 6/11 in papillomas from 3 patients, and HPV types 6/11, 16/18, and 31/33/51 in a papilloma of a fourth patient. No HPV DNA was detected in papillomas of 2 patients. We found HPV 6/11 in 4 of the carcinomas. We conclude that the spontaneous transformation of RRP to SCCA is not characterized by a histologic progression through dysplasia over time. Transformation can result in the loss of HPV expression. It does not appear that p53 is a molecular marker for monitoring the transformation process. Thus, these cancers may be very difficult to diagnose histologically and clinically early in the course of the transformation of the disease.

CO(2) laser treatment in 244 patients with respiratory papillomas.

OBJECTIVE: Respiratory papillomas (RP) tend to recur and the difficulty in eradicating the disease makes their treatment frustrating. Meticulous CO(2) laser excisions every 2 months has been the most effective treatment to date. This article analyzes the results of this plan in 244 patients with RP in the nose, nasopharynx, pharynx, hypopharynx, larynx, trachea, lung parenchyma, and skin. METHODS: Two hundred forty-four patients with recurrent RP were treated by the senior author with CO(2) laser excisions and, in some cases, podophyllum and alpha interferon. Demographics, initial distribution of papillomas, number of operations performed on each patient, and current results were evaluated. RESULTS: Careful laser excisions of RPs every 2 months achieved "remission" of disease (no visible RP on indirect or often direct laryngoscopy 2 mo after last removal) in 37% of patients, "clearance" of the disease process (no RP clinically apparent for 3 y after last removal) in 6%, and "cure" (no clinical recurrence for 5 y after last removal) in 17%. Juvenile-onset RP tends to follow a more aggressive course than adult-onset RP. Four patients (1.6%) developed malignant transformation of their papillomas. Except for ones in lung parenchyma, RP in areas other than the true vocal cords tend to be cleared faster because aggressive removal does not cause hoarseness. Lung parenchyma RPs are eventually fatal because of pulmonary failure from abscesses and cysts resulting from a lack of effective treatment. CONCLUSION: Frequent and meticulously performed CO(2) laser excisions can achieve significant voice and airway improvement, and some clinical "cures." However, effective antiviral medicines and/or immunologic agents are needed to achieve true cures with elimination of all human papilloma virus 6 and 11 viruses.

Association of Kaposi's sarcoma associated herpesvirus (KSHV) DNA in bronchoalveolar lavage fluid of HIV infected individuals with bronchoscopically diagnosed tracheobronchial Kaposi's sarcoma.

OBJECTIVES: To determine the frequency of detection of Kaposi's sarcoma associated herpesvirus (KSHV), also known as human herpesvirus (HHV) type 8, DNA in bronchoalveolar lavage (BAL) fluid from HIV infected individuals with and without KS and to compare this with the detection rate in peripheral blood. Also to identify whether KSHV was associated with specific cell types in lavage fluid. METHODS: Nested PCR was used to detect KSHV DNA in BAL fluid from 41 consecutive individuals with Kaposi's sarcoma (KS) and in 41 controls with similar CD4 lymphocyte counts. Semiquantification of viral DNA was by end point titration. A positive cell sorting selection procedure was used to isolate specific BAL fluid cell types. RESULTS: KSHV DNA was detected in BAL fluid from 24 of 29 (83%) individuals with a bronchoscopic diagnosis of tracheobronchial KS. None was detected in 12 individuals with only cutaneous KS, or in 41 matched controls without KS. In five, KSHV DNA was detected in the cell depleted and cellular fractions of BAL fluid and in 1/5 in the CD14 (macrophage) fractions. None was detected in the CD19 (B lymphocyte) or CD4/CD8 (T lymphocyte) fractions. CONCLUSIONS: There was a clear association between the diagnosis of tracheobronchial KS and detection of KSHV DNA in BAL fluid. The cell type supporting KSHV in the respiratory tract is not CD 19 positive and has yet to be conclusively identified.

Pediatric respiratory papillomatosis: prognostic role of viral typing and cofactors.

Children with recurrent respiratory papillomatosis vary greatly in their clinical disease course. Many have mild disease with eventual remission while others present with an early aggressive airway obstructive course. This study consisted of 24 pediatric patients whose specimens underwent polymerase chain reaction analysis for cytomegalovirus (CMV), herpes simplex virus (HSV), and human papillomavirus (HPV) type. Nineteen of 24 specimens contained enough DNA for this study. None of the specimens were found to contain DNA from HPV-16, -18, -31, -33; CMV; or HSV, which contrasts with our previous findings in adults. Ten patients were infected by HPV-11 and seven of these underwent tracheotomy because of an aggressive tumorigenic clinical course. Nine patients were infected by HPV-6 alone of whom only two required a tracheotomy (P = 0.05, Fisher's Exact Test). The early airway obstructive course associated with HPV-11, however, had no bearing on achieving eventual disease remission, with decannulation achieved in eight of nine children.

Clinicopathological consultation. Lymphoepithelial carcinoma of the larynx hypopharynx, and trachea.

Lymphoepithelial carcinoma of the larynx, hypopharynx, and trachea is a rare neoplasm composed of large, poorly differentiated, nonkeratinized cells intermingled with small nonneoplastic lymphocytes and plasma cells. It is histologically similar to its more common counterpart occurring in the nasopharynx. In contrast to nasopharyngeal carcinoma, most cases have not been associated with Epstein-Barr virus (EBV), although rare cases have been reported to be EBV-positive. The diagnosis often requires immunohistochemistry or electron microscopy for confirmation. The neoplasm seems to behave in a fashion reminiscent of nasopharyngeal carcinoma. Lymph node metastasis occurs in the majority of patients, and eventual visceral dissemination occurs in one fourth. Radiotherapy is the main treatment for the primary tumor and regional metastases, but chemotherapy is indicated for more advanced disease. The initial stage is the primary determinant of prognosis. Death from disease occurs in about one third of patients.