Bettie M. Steinberg

Laryngeal Papillomavirus Infection during Clinical Remission

Biopsy samples from 20 patients with a history of laryngeal papillomas were analyzed by Southern blot hybridization for the presence of human papillomavirus DNA. All papillomas examined contained papillomavirus DNA sequences. Four samples from uninvolved sites in two patients with active disease and eight samples from patients in remission also contained viral DNA. No human papillomavirus DNA was detected in a third patient seen during remission. These results explain the clinical pattern of frequent recurrences even after long-term remission in laryngeal papillomatosis.

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.

The natural history of human papillomavirus infections of the mucosal epithelia

Chow LT, Broker TR, Steinberg BM. The natural history of human papillomavirus infections of the mucosal epithelia. APMIS 2010; 118: 422–449.

Recurrent respiratory papillomatosis: a complex defect in immune responsiveness to human papillomavirus‐6 and ‐11

Bonagura VR, Hatam LJ, Rosenthal DW, DeVoti JA, Lam F, Steinberg BM, Abramson AL. Recurrent respiratory papillomatosis: a complex defect in immune responsiveness to human papillomavirus‐6 and ‐11. APMIS 2010; 118: 455–470.

A possible role for human papillomaviruses in head and neck cancer

Human papillomaviruses (HPVs) cause benign tumors in the respiratory tract. Mounting evidence suggests that they also play a role in the etiology of a subset of head and neck cancers. Carcinomas in patients with a history of recurrent respiratory papillomatosis clearly are caused by persisting HPV interacting with one or more carcinogenic agents. Verrucous carcinomas of the oral cavity, tonsillar and tongue carcinomas are strongly linked with HPVs, based on molecular epidemiologic data. Tonsillar cancers have been shown to express HPV RNA, presumed necessary to induce and maintain a carcinoma, supporting a viral etiology. This paper reviews the molecular and cellular basis for considering HPVs as causttive agents of cancer, and reviews the literature that considers the possible role of HPVs in head and neck cancer.

Efficacy of DHE photodynamic therapy for respiratory papillomatosis: immediate and long-term results.

Objectives/Hypothesis: Recurrent respiratory papillomatosis is a potentially life‐threatening disease that affects both children and adults and can result in complete respiratory obstruction. Conventional therapies cannot prevent multiple recurrences. The authors have been evaluating photodynamic therapy (PDT) to treat this disease since 1988. This study compared the efficacy of PDT with dihematoporphyrinether (DHE) with traditional therapy. Study Design: This was a randomized prospective trial of DHE‐PDT. Patients were randomly assigned to receive one of two doses of DHE—3.25 mg/kg or 4.25 mg/kg body weight. They were compared with a concurrent control group. Disease extent was evaluated by direct laryngoscopy before treatment and over a 1‐year period following treatment. Results were also compared with two historical cohorts of patients treated with lower doses of DHE. Methods: Eighty‐one patients, ages 4 to 74 years, with moderate to severe recurrent disease were enrolled. Forty‐eight received PDT and 33 in the control group were treated with conventional therapy. Both PDT groups received 50 J laser light to activate the drug. Patients received an intravenous infusion of DHE as outpatients 48 to 72 hours before treatment. During direct laryngoscopy, light (630 nm) was delivered by an argon‐pumped dye laser. Tissue biopsies were analyzed for presence of human papillomavirus (HPV). Results: There was notable improvement with either drug dose over the first year. Those receiving 4.25 mg/kg DHE experienced a significantly larger decrease in papilloma growth rate. Three‐year follow‐up of a subset of patients confirmed that improvement was maintained. There was no impact of DHE‐PDT on persistence of HPV DNA. Conclusion: This therapy holds promise for the treatment of laryngeal papillomas.

PTEN is a negative regulator of STAT3 activation in human papillomavirus-infected cells.

Laryngeal papillomas are caused by infection of the laryngeal epithelium by human papillomavirus type 6 or type 11 (HPV-6/-11). Previous studies in our laboratory have demonstrated an increase in PI3 kinase levels in papilloma tissue. However, activation of the downstream effector of PI3 kinase, protein kinase B (PKB/Akt), was reduced. This observation was explained by the elevated expression of the phosphatase and tensin homologue (PTEN), a recently characterized tumour suppressor, in papilloma tissue. Recent investigation of the possible functional roles of PTEN during papilloma development has now indicated that the level of tyrosine(705)-phosphorylated signal transducer and activator of transcription 3 [PTyr(705)STAT3] could be inversely correlated to that of PTEN as well. In vitro phosphatase assays suggested the presence of an increased level of a PTyr(705)STAT3 phosphatase in papilloma extract. Immunodepletion of PTEN from papilloma extracts resulted in a reduction of the PTyr(705)STAT3 phosphatase activity. Transfection of PTEN cDNA into HeLa cells attenuated STAT3 phosphorylation at Tyr(705) in a dose-dependent manner. This attenuation of STAT3 phosphorylation was independent of the STAT3 kinase. Interestingly, introduction of a lipid phosphatase mutant of PTEN (G129E) resulted in heightened PTyr(705)STAT3 phosphatase activity, relative to that obtained from wild-type PTEN transfection. These data indicate that PTEN negatively regulates STAT3 activation in HPV-infected papilloma cells. Induction of PTEN and reduction of activated STAT3 might be a result of a host defence mechanism or a virus-directed strategy to alter normal epithelial differentiation programming.

Isolation and characterization of T antigen-negative revertants from a line of transformed rat cells containing one copy of the SV40 genome

Negative selection with FUdR produced revertants from the transformed rat line 14B, which contains one insertion of the SV40 viral genome (Botchan, Topp and Sambrook, 1976). 14B contains nuclear T antigen, grows to a high density, grows in low serum and is anchorage-independent. The revertants fall into three classes with regard to viral DNA sequences: the SV40 DNA is retained; the SV40 DNA is retained but has undergone a deletion; and the SV40 DNA is lost, generating a cured cell. This heterogeneity is not a result of long-term passage. The revertants arise with a frequency of one in 8.4 X 10(5) cells after as few as 12 passages. All three classes of revertants are T antigen-negative, density-sensitive, more serum sensitive than 14B and anchorage-dependent. These data argue for a direct role of the functioning viral genome in the maintenance of the transformed state, and that with 14B, the phenotypes of transformation are not virus gene dosage-dependent.

Human papillomavirus 6/11 and 16/18 in schneiderian inverted papillomas. In situ hybridization with human papillomavirus RNA probes

Schneiderian inverted papillomas may be troublesome lesions for clinicians with propensity for recurrences. Dysplasia is not uncommonly seen, and some of these lesions do progress to develop squamous carcinoma. The authors hybridized in situ seven inverted papillomas with RNA probes to human papillomavirus (HPV) 6, 11, 16, and 18. Four of these contained dysplasia, two were without dysplasia, and one contained invasive squamous cell carcinoma. Five inverted papillomas showed evidence of HPV infection based on hybridization. One with mild to moderate and one with severe dysplasia, and one without dysplasia hybridized with mixed probe HPV 6/11. One with mild dysplasia and one associated with invasive squamous cell carcinoma hybridized with mixed probe HPV 16/18. One inverted papilloma without dysplasia and one with severe dysplasia did not definitively hybridize with either mixed probe. These findings raise interesting questions as to role of HPV 6/11 and 16/18 in the development of inverted papillomas, and probably on the progression to and carcinoma.

Human papillomavirus type 6a DNA in the lung carcinoma of a patient with recurrent laryngeal papillomatosis is characterized by a partial duplication

Transcriptionally active human papillomavirus type 6a (HPV-6a) DNA was detected in a lung carcinoma of a patient with recurrent laryngeal papillomatosis. The carcinoma contained episomal HPV-6a genomes that had a duplication of the upstream regulatory region, the late region and a portion of the early region. HPV-6a genomes found in benign laryngeal papillomas from the same patient did not contain this duplication. A role for the mutant molecules in the pathogenesis of the malignancy is suggested.