Spontaneous and Vaccine-Induced Clearance of Mus Musculus Papillomavirus 1 Infection.

Mus musculus papillomavirus 1 (MmuPV1/MusPV1) induces persistent papillomas in immunodeficient mice but not in common laboratory strains. To facilitate the study of immune control, we sought an outbred and immunocompetent laboratory mouse strain in which persistent papillomas could be established. We found that challenge of SKH1 mice (Crl:SKH1-Hrhr) with MmuPV1 by scarification on their tail resulted in three clinical outcomes: (i) persistent (>2-month) papillomas (∼20%); (ii) transient papillomas that spontaneously regress, typically within 2 months (∼15%); and (iii) no visible papillomas and viral clearance (∼65%). SKH1 mice with persistent papillomas were treated by using a candidate preventive/therapeutic naked-DNA vaccine that expresses human calreticulin (hCRT) fused in frame to MmuPV1 E6 (mE6) and mE7 early proteins and residues 11 to 200 of the late protein L2 (hCRTmE6/mE7/mL2). Three intramuscular DNA vaccinations were delivered biweekly via in vivo electroporation, and both humoral and CD8 T cell responses were mapped and measured. Previously persistent papillomas disappeared within 2 months after the final vaccination. Coincident virologic clearance was confirmed by in situ hybridization and a failure of disease to recur after CD3 T cell depletion. Vaccination induced strong mE6 and mE7 CD8+ T cell responses in all mice, although they were significantly weaker in mice that initially presented with persistent warts than in those that spontaneously cleared their infection. A human papillomavirus 16 (HPV16)-targeted version of the DNA vaccine also induced L2 antibodies and protected mice from vaginal challenge with an HPV16 pseudovirus. Thus, MmuPV1 challenge of SKH1 mice is a promising model of spontaneous and immunotherapy-directed clearances of HPV-related disease.IMPORTANCE High-risk-type human papillomaviruses (hrHPVs) cause 5% of all cancer cases worldwide, notably cervical, anogenital, and oropharyngeal cancers. Since preventative HPV vaccines have not been widely used in many countries and do not impact existing infections, there is considerable interest in the development of therapeutic vaccines to address existing disease and infections. The strict tropism of HPV requires the use of animal papillomavirus models for therapeutic vaccine development. However, MmuPV1 failed to grow in common laboratory strains of mice with an intact immune system. We show that MmuPV1 challenge of the outbred immunocompetent SKH1 strain produces both transient and persistent papillomas and that vaccination of the mice with a DNA expressing an MmuPV1 E6E7L2 fusion with calreticulin can rapidly clear persistent papillomas. Furthermore, an HPV16-targeted version of the DNA can protect against vaginal challenge with HPV16, suggesting the promise of this approach to both prevent and treat papillomavirus-related disease.

Risk Factors for Dysplasia in Recurrent Respiratory Papillomatosis in an Adult and Pediatric Population.

AIM: Recurrent respiratory papillomatosis (RRP) is classically described as a benign neoplasm of the larynx caused by the low-risk human papillomavirus (HPV) viral subtypes. Nevertheless, transformation to dysplasia and invasive carcinoma can occur. We aimed to assess the prevalence of dysplasia and carcinoma-ex-papilloma in both adult-onset and juvenile-onset RRP and identify patient risk factors for this dysplastic transformation. MATERIAL AND METHODS: Ten-year retrospective chart review of a tertiary otolaryngology referral center. Patients with papilloma were identified from a review of a pathology database and clinical records. Patient demographics, pathologic data, and treatment history, including use of cidofovir as an adjunctive therapy for papilloma, were extracted from electronic medical records. RESULTS: One hundred fifty-nine RRP patients were identified, 96 adult-onset (AORRP) and 63 juvenile-onset (JORRP) cases. Of this cohort, 139 (87%) had only benign papilloma as a pathologic diagnosis. In the AORRP cohort, 10 patients (10%) were diagnosed with dysplasia or carcinoma in situ in addition to papilloma, and 5 patients (5%) had malignant transformation to invasive carcinoma-ex-papilloma. There was a significantly higher age of disease onset for those with dysplasia or carcinoma versus those without dysplasia or carcinoma (56 vs 45 years old; P = .0005). Of the 63 JORRP patients, there were no cases of dysplasia but 3 (5%) cases of invasive carcinoma-ex-papilloma, all involving pulmonary disease. The JORRP patients with carcinoma-ex-papilloma had a younger average disease onset (2 vs 6 years old; P = .009) and a higher rate of tracheal involvement than those without carcinoma. Gender, smoking history, number of operations, or use of cidofovir showed no association with the development of dysplasia or carcinoma-ex-papillomatosis in either the AORRP or JORRP population. CONCLUSION: In a large series of RRP, age of disease onset is the strongest predictor of dysplastic transformation in the adult and pediatric population. Carcinoma-ex-papillomatosis was uniformly associated with pulmonary disease in the JORRP population in this series. No other demographic or behavioral factors, including adjunctive therapy with cidofovir, were statistically associated with dysplasia or carcinoma-ex-papilloma.

Predictors of Posterior Glottic Stenosis: A Multi-Institutional Case-Control Study.

OBJECTIVE: To assess intrinsic and extrinsic risk factors in the development of posterior glottic stenosis (PGS) in intubated patients. METHODS: Patients diagnosed with PGS between September 2012 and May 2014 at 3 tertiary care university hospitals were included. Patient demographics, comorbidities, duration of intubation, endotracheal tube (ETT) size, and indication for intubation were recorded. Patients with PGS were compared to control patients represented by patients intubated in intensive care units (ICU). RESULTS: Thirty-six PGS patients were identified. After exclusion, 28 PGS patients (14 male, 14 female) and 112 (65 male, 47 female) controls were studied. Multivariate analysis demonstrated ischemia (P < .05), diabetes (P < .01), and length of intubation (P < .01) were significant risk factors for the development of PGS. Fourteen of 14 (100%) males were intubated with a size 8 or larger ETT compared to 47 of 65 (72.3%) male controls (P < .05). Posterior glottic stenosis (P < .01), length of intubation (P < .001), and obstructive sleep apnea (P < .05) were significant risk factors for tracheostomy. CONCLUSION: Duration of intubation, ischemia, diabetes mellitus, and large ETT size (8 or greater) in males were significant risk factors for the development of PGS. Reducing the use of size 8 ETTs and earlier planned tracheostomy in high-risk patients may reduce the incidence of PGS and improve ICU safety.

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.

Dysphagia, short-term outcomes, and cost of care after anterior cervical disc surgery.

Dysphonia and dysphagia are common complications of anterior cervical discectomy (ACD). We sought to determine the relationship between dysphagia and in-hospital mortality, complications, speech therapy/dysphagia training, length of hospitalization, and costs associated with ACD. Discharge data from the Nationwide Inpatient Sample for 1,649,871 patients who underwent ACD of fewer than four vertebrae for benign acquired disease between 2001 and 2010 were analyzed using cross-tabulations and multivariate regression modeling. Dysphagia was reported in 32,922 cases (2.0 %). Speech therapy/dysphagia training was reported in less than 0.1 % of all cases and in only 0.2 % of patients with dysphagia. Dysphagia was significantly associated with age ≥65 years (OR = 1.5 [95 % CI 1.4-1.7], P < 0.001), advanced comorbidity (OR = 2.3 [2.0-2.6], P < 0.001), revision surgery (OR = 2.7 [2.3-3.1], P < 0.001), disc prosthesis placement (OR = 1.5 [1.0-2.0], P = 0.029), and vocal cord paralysis (OR = 11.6 [8.3-16.1], P < 0.001). Dysphagia was a significant predictor of aspiration pneumonia (OR = 8.6 [6.7-10.9], P < 0.001), tracheostomy (OR = 2.3 [1.6-3.3], P < 0.001), gastrostomy (OR = 30.9 [25.3-37.8], P < 0.001), and speech therapy/dysphagia training (OR = 32.0 [15.4-66.4], P < 0.001). Aspiration pneumonia was significantly associated with in-hospital mortality (OR = 15.9 [11.0-23.1], P < 0.001). Dysphagia, vocal cord paralysis, and aspiration pneumonia were significant predictors of increased length of hospitalization and hospital-related costs, with aspiration pneumonia having the single largest impact on length of hospitalization and costs. Dysphagia is significantly associated with increased morbidity, length of hospitalization, and hospital-related costs in ACD patients. Despite the known risk of dysphagia in ACD patients and an established role for the speech-language pathologist in dysphagia management, speech-language pathology intervention appears underutilized in this population.

Posterior Glottic Injury Following Prolonged Intubation in COVID-19 Patients.

OBJECTIVE: The objective of this study was to characterize the risk factors for posterior glottic injury (PGI) in patients with coronavirus disease 2019 (COVID-19) who underwent prolonged intubation. STUDY DESIGN: This was a case-control study designed to assess the risk factors associated with development of PGI in COVID-19 patients who underwent prolonged intubation. SETTING: This single-center study was conducted at a tertiary care academic hospital in a metropolitan area. METHODS: We retrospectively reviewed patients who underwent prolonged intubation (≥7 days) for COVID-19 and compared those with PGI to those without. Patient demographics, comorbidities, and intubation characteristics were compared. Factors associated with PGI development among COVID-19 patients were assessed using multivariate regression. RESULTS: We identified 56 patients who presented with PGI following prolonged intubation for COVID-19 and 60 control patients who underwent prolonged intubation for COVID-19 but did not develop PGI. On univariate analyses, the number of reintubations due to failed extubation efforts was significantly associated with development of PGI (odds ratio [OR], 2.9; 95% CI, 1.4-6.2). On multivariate analyses, patients with cardiovascular disease (OR, 3.3; 95% CI, 1.2-9.0); non-COVID-19 respiratory illnesses, which included obstructive sleep apnea and asthma (OR, 5.9; 95% CI, 2.0-17.8); and diabetes mellitus (OR, 11.6; 95% CI, 3.7-36.6) were more likely to develop PGI. CONCLUSION: Our results represent the largest case-control study investigating risk factors for PGI in the setting of prolonged intubation specific to COVID-19. Our study suggests a significant role of comorbidities associated with poor wound healing with development of PGI.

Impact of human immunodeficiency virus status on laryngeal cancer survival and locoregional control.

OBJECTIVES: To compare long-term outcomes of laryngeal cancer (LC) in people living with HIV (PLWH) versus uninfected individuals and determine how clinical and viral factors-such as demographics, cancer stage, HIV viral load, and CD4 nadir-contribute to these outcomes. METHODS: This was a retrospective case-control study of 749 patients seen for LC at a single tertiary care center between 2003 and 2017. Of these, 22 had HIV at the time of LC diagnosis, and they were matched in a 1:4 ratio to uninfected controls based on sex, presence of smoking history, and age at cancer diagnosis. Kaplan-Meier survival curves and Cox proportional hazards models were constructed to identify overall and disease-free survival differences based on HIV status, as well as other clinical and viral factors. RESULTS: Compared to all uninfected individuals, PLWH were diagnosed with LC approximately 6 years younger (p = .013). 1-, 2-, and 5-year overall survival for PLWH were 86.4% (63.4%-95.4%), 77.3% (53.7%-89.9%), and 65.8% (40.8%-82.2%), respectively following LC diagnosis, and HIV was not significantly associated with overall (HR = 3.34 [0.59-18.79]) or disease-free survival (HR = 2.12 [0.71-6.36]). The incidence rate of locoregional recurrence among PLWH was 541 compared to 371 per 10,000 person-years in controls, which were not significantly different (p = .420). Furthermore, among PLWH, peak viral load and CD4 nadir were not associated with overall or disease-free survival. CONCLUSION: While previous work has shown that HIV is associated with elevated risk of LC, survival did not differ significantly between PLWH and uninfected individuals in this study. LEVEL OF EVIDENCE: 3.

Geriatric Dysphonia: Characteristics of Diagnoses in Age-Based Cohorts in a Tertiary Voice Clinic.

OBJECTIVE: An aging population has increased focus on geriatric otolaryngology. Those ≥65 years old are not a uniform population, however, and recent gerontology literature recognizes important physiologic differences between the young-old (ages 65-74 years), middle-old (75-84), and old-old (≥85). This study evaluates differences within these groups among dysphonia patients ≥65 years relative to diagnosis and voice-related quality of life (V-RQOL). METHODS: Chart review of all new patients ≥65 years presenting to the Johns Hopkins Voice Center between April 2015 and March 2017 identified chief complaint, diagnosis, and self-reported voice handicap. Etiology of dysphonia diagnoses were classified. Diagnostic categories and V-RQOL were evaluated as functions of patient age and gender. RESULTS: Of 839 new patients ≥65 years, 463 (55.2%) reported chief complaint of dysphonia, with the most common etiologies being vocal fold immobility (28.3%) and atrophy (21.6%). Younger cohorts were more likely to present with benign vocal fold lesion and vocal fold immobility, while older cohorts were more likely to present with atrophy ( P = .016). The odds of having a diagnosis of vocal fold atrophy increased 7% with each year of life (odds ratio = 1.07; 95% CI, 1.03-1.11). V-RQOL scores were similar across gender and age categorization. CONCLUSION: Dysphonia patients ≥65 years are not a uniform group, and important differences exist in terms of diagnosis as a function of age. Knowledge of these differences may inform further investigations in the growing field of geriatric otolaryngology.

The PD-1 and PD-L1 pathway in recurrent respiratory papillomatosis.

OBJECTIVES/HYPOTHESIS: Generation of an immunosuppressive microenvironment may enable a persistent human papillomavirus infection in the setting of an otherwise normal immune system. We hypothesized that expression of the T-lymphocyte co-inhibitory receptor programmed death 1 (PD-1) and its ligand PD-L1 would be increased in the recurrent respiratory papillomatosis (RRP) microenvironment compared to normal controls. STUDY DESIGN: Case-control study. METHODS: Formalin-fixed paraffin-embedded respiratory papilloma and normal controls were obtained under institutional review board approval, stained for CD4, CD8, FoxP3, and PD-1, and scored by automated cell count. PD-L1 staining was scored by a blinded pathologist using an adjusted inflammation score that accounted for epithelial and immune infiltrate. RESULTS: Thirty-nine RRP cases and seven controls were studied. All immunologic markers demonstrated significantly increased staining in RRP specimens compared to normal controls (all P < .01). PD-1 correlated with both CD4 (P < .0001) and CD8 (P < .001) cell counts. Epithelial staining for PD-L1 (68%) and PD-L1+ infiltrating immune cells (76%) were observed in the majority of papilloma samples. The strongest staining for PD-L1 was usually observed in the basal papilloma layer adjacent to the immunologic infiltrate in the vascular core. Disease severity inversely correlated with CD8 cell counts (P = .01). A correlation between disease severity and other immunologic markers was not observed. CONCLUSIONS: Most RRP specimens demonstrate PD-1 T-lymphocyte infiltration and PD-L1 expression on both papilloma and infiltrating immune cells. This study suggests that this checkpoint pathway may be contributing to local immunosuppression in RRP, and opens the door for clinical trials utilizing PD-blocking monoclonal antibodies. LEVEL OF EVIDENCE: NA Laryngoscope, 128:E27-E32, 2018.

The Changing Impact of Gastroesophageal Reflux Disease in Clinical Practice.

OBJECTIVES: The National Ambulatory Medical Care Survey (NAMCS) database was utilized to understand evolving national trends in diagnosis and management of reflux. METHODS: The NAMCS database was queried for visits related to gastroesophageal reflux diagnosis and management. Analysis performed for time periods 1998-2001, 2002-2005, and 2006-2009 was weighted to provide national estimates of care. Results were compared to previously reported time periods from 1990 to 2001 to evaluate patterns in overall visits, age and ethnicity of patients, provider type, and prescriptions provided. RESULTS: The number of ambulatory visits for reflux increased from 8 684 000 in 1998-2001 to 15 750 000 in 2006-2009. Visits increased across each time period for internal medicine, family, and gastroenterology physicians. Among otolaryngologists, absolute visits increased from 1998-2001 to 2002-2005 but decreased in 2006-2009; difference between these time periods did not reach statistical significance. From 1998-2001 to 2006-2009, reflux medication use increased 233%, with continuing trends toward increased proton pump inhibitor use. CONCLUSIONS: Reflux visits have increased across all demographic subgroups studied. Knowledge of these trends may inform further paradigm shifts in diagnosis and management of reflux.

Evaluation of Dyspnea Outcomes After Endoscopic Airway Surgery for Laryngotracheal Stenosis.

Importance: Endoscopic airway surgery is a frequently used procedure in the management of laryngotracheal stenosis (LTS); however, no established outcome measures are available to assess treatment response. Objective: To assess acoustics and aerodynamic measures and voice- and dyspnea-related quality of life (QOL) in adult patients with LTS who undergo endoscopic airway surgery. Design, Setting, and Participants: This case series compared preoperative measures and postoperative outcomes among adult patients who underwent endoscopic airway surgery for LTS from September 1, 2013, to September 30, 2015, at the tertiary care Johns Hopkins Voice Center. Patients were excluded if they did not undergo balloon dilation or if they had multilevel or glottic stenosis. The Phonatory Aerodynamic System was used to quantify laryngotracheal aerodynamic changes after surgery. Final follow-up was completed 2 to 6 weeks after surgery. Main Outcomes and Measures: The voice-related QOL instrument (V-RQOL), Dyspnea Index, and Clinical Chronic Obstructive Pulmonary Disease Questionnaire were completed before and after endoscopic surgery. Consensus auditory perceptual evaluation of voice, acoustic measurements, and aerodynamic outcomes were also assessed. Results: Fourteen patients (1 man and 13 women; mean [SD] age, 45.4 [4.3] years) were enrolled. The mean postoperative V-RQOL scores (n = 14) increased from 74.3 to 85.5 (mean of difference, 11.3; 95% CI, 2.2 to 20.3). The mean postoperative Dyspnea Index (n = 14) decreased from 26.9 to 6.6 (mean of difference, -20.3; 95% CI, -27.9 to -12.7); the mean postoperative Clinical Chronic Obstructive Pulmonary Disease Questionnaire scores (n = 9) decreased from 3.2 to 1.0 (mean of difference, -2.2; 95% CI, -3.4 to -0.9). Postoperative mean vital capacity (n = 14) increased from 2.5 to 3.1 L (mean of difference, 0.6 L; 95% CI, 0.3-1.0 L), whereas mean laryngeal resistance (n = 14) decreased from 73.9 to 46.4 cm H2O/L/s (mean of difference, -27.5 cm H2O/L/s; 95% CI, -44.8 to -10.3 cm H2O/L/s) postoperatively. Conclusions and Relevance: Patients demonstrate statistically and clinically significant improvement in dyspnea-related QOL, whereas a few patients showed a clinically significant improvement in V-RQOL. Dyspnea-related QOL outcomes should be added to airway surgeons' regular assessment of patients with LTS to measure treatment response and inform the decision to perform a second operation, whereas V-RQOL outcomes need additional prospective study with a larger sample size. The Phonatory Aerodynamic System is not an optimal method to quantify changes in laryngotracheal aerodynamics after intervention in LTS.

Patient-perceived long-term communication and swallow function following cerebellopontine angle surgery.

OBJECTIVES/HYPOTHESIS: Evaluation of long-term patient-perceived functional outcomes and quality of life (QOL) related to communication and eating with an emphasis on voice, speech production, and swallowing after cerebello-pontine angle (CPA) surgery. STUDY DESIGN: Prospective cross-sectional study. METHODS: The MD Anderson Dysphagia Inventory (MDADI), Voice Handicap Index (VHI), and Facial Clinimetric Evaluation (FaCE) surveys were distributed to patients who underwent CPA surgery between January 2008 and December 2010. Immediate postoperative cranial nerve function extracted from medical records was compared to long-term patient-perceived function and associated QOL. RESULTS: There was a 61% response rate with a mean postoperative period of 31.6 months (range 15-49). The presence of facial palsy in the postoperative period and the corresponding House-Brackmann (H-B) score were the strongest predictors of patient-perceived long-term function and QOL in all three domains (P < .005). Postoperative vagal palsy by comparison was not associated with long-term disturbance of voice or speech function. Postoperative dysphagia had a particularly large association with perceived long-term facial function and related QOL (P < .0005), with a smaller but significant impact on perceived swallow outcome (P < .05). After adjusting for other variables, the postoperative H-B score remained a significant predictor of perceived long-term facial and voice function and related QOL. CONCLUSIONS: Patients with severe facial dysfunction following surgery to the CPA are at increased risk for long-term self-reported difficulties with communication and eating, even with improvement of vagal function. Speech and swallow therapy should therefore be provided to these patients whether or not they also have pharyngeal dysphagia or voice disturbance. LEVEL OF EVIDENCE: 2b.