Papilloma Tissue Research Articles

Characterization of HPV6/11-reactive T-cell subsets in papillomas of patients with juvenile-onset recurrent respiratory papillomatosis and identification of HPV11 E7-specific candidate TCR clonotypes.

Juvenile-onset recurrent respiratory papillomatosis (JORRP) is caused by persistent infection of epithelial cells by low-risk human papillomavirus (HPV) types 6 and 11. While multiple infiltrated immune cells have been reported to mediate disease progress, knowledge of HPV-reactive T-cell subsets in papillomas remains elusive. Through single-cell RNA sequencing and RNA microarray, we found that CD8+ tissue-resident memory T (CD8+ TRM) cells with strong interferon-gamma (IFN-γ) production expanded, and were negatively correlated to the disease severity in the frequency of surgery. These IFN-γ+ CD8+ memory T cells were readily activated and expanded in vitro by autologous dendritic cells loaded with HPV11 E7 peptide pool. Moreover, T cell receptor (TCR) clonal expansion was observed in JORRP papilloma tissues, indicating a biased TCR repertoire toward HPV-specific recognition. Finally, we identified and characterized HPV11 E7-specific candidate TCR clonotypes from IFN-γ+ CD8+ memory T cells, suggesting their potential application in TCR-engineered T cells (TCR-T) therapy for HPV11-related diseases. Our findings provided insights into the specific local immune response to HPV6/11 infection and highlighted the importance of IFN-γ+ CD8+ TRM cells in anti-HPV6/11 T-cell immunity.IMPORTANCEThe persistent recurrence of human papillomavirus (HPV) 6/11 infection in papillomas underscores the failure of local immune responses in patients with juvenile-onset recurrent respiratory papillomatosis (JORRP). Our previous study demonstrated that T cells constitute the predominant immune cell population in JORRP papilloma tissues. Understanding the T-cell-mediated immune responses within JORRP papilloma tissues is crucial for disease control. In the present study, we characterized CD8+ tissue-resident memory T (CD8+ TRM) cells as the primary T-cell subset responsible for local anti-HPV6/11 immunity. Moreover, we identified two HPV11 E7-specific candidate T cell receptor (TCR) clonotypes out of IFN-γ+ CD8+ memory T cells. Overall, our findings provided insights into the local immune responses to HPV6/11 infection and offered information for developing more effective immunotherapeutic strategies against JORRP.

Deep learning model for differentiating nasal cavity masses based on nasal endoscopy images

BackgroundNasal polyps and inverted papillomas often look similar. Clinically, it is difficult to distinguish the masses by endoscopic examination. Therefore, in this study, we aimed to develop a deep learning algorithm for computer-aided diagnosis of nasal endoscopic images, which may provide a more accurate clinical diagnosis before pathologic confirmation of the nasal masses.MethodsBy performing deep learning of nasal endoscope images, we evaluated our computer-aided diagnosis system’s assessment ability for nasal polyps and inverted papilloma and the feasibility of their clinical application. We used curriculum learning pre-trained with patches of nasal endoscopic images and full-sized images. The proposed model’s performance for classifying nasal polyps, inverted papilloma, and normal tissue was analyzed using five-fold cross-validation.ResultsThe normal scores for our best-performing network were 0.9520 for recall, 0.7900 for precision, 0.8648 for F1-score, 0.97 for the area under the curve, and 0.8273 for accuracy. For nasal polyps, the best performance was 0.8162, 0.8496, 0.8409, 0.89, and 0.8273, respectively, for recall, precision, F1-score, area under the curve, and accuracy. Finally, for inverted papilloma, the best performance was obtained for recall, precision, F1-score, area under the curve, and accuracy values of 0.5172, 0.8125, 0.6122, 0.83, and 0.8273, respectively.ConclusionAlthough there were some misclassifications, the results of gradient-weighted class activation mapping were generally consistent with the areas under the curve determined by otolaryngologists. These results suggest that the convolutional neural network is highly reliable in resolving lesion locations in nasal endoscopic images.

ЭТИОЛОГИЯ СИНОНАЗАЛЬНОЙ ИНВЕРТИРОВАННОЙ ПАПИЛЛОМЫ: СОВРЕМЕННЫЙ ВЗГЛЯД НА ПРОБЛЕМУ

Abstract. Introduction. Inverted papilloma is the most common type of sinonasal papilloma. It refers to benign tumors, but its clinical course is often associated with post-surgical recurrences and malignancy. Although the morphological characteristics and clinical features of inverted papilloma are well known, its etiology and risk factors remain a matter of debate. Aim. The aim of the study was to analyze the contemporary literature and summarize the data available regarding the significance of viral infection, cell cycle regulator proteins, angiogenesis, chronic inflammation, environmental influences, and other factors for the risk of inverted papilloma. Materials and Methods. A search was made in databases, such as SCOPUS, PubMed, Google Scholar, and RSCI, using the following keywords: Sinonasal papilloma, inverted nasal papilloma, Schneiderian papilloma. Results and Discussion. Human papillomavirus is considered the main growth trigger of inverted papilloma. Potential role of human papillomavirus in initiating the tumor growth is related to the presence in its genome of genes encoding the E6 and E7 proteins. Certain cell cycle regulatory factors and angiogenic proteins contribute to the dysregulation of proliferation and apoptosis and facilitate cell migration and tumor invasion. However, despite the frequent detection of human papillomavirus DNA and associated transcription factors in inverted papilloma tissues, significance of the papillomavirus infection in the occurrence of inverted papilloma or its transformation into malignant forms has not been proven reliably yet. It should be noted that human papillomavirus stimulates not only apoptosis inhibition and the uncontrolled division of epithelial cells, but also angiogenesis, which is also a tumor growth trigger. Among other pathogenic factors, we also discussed the importance of chronic inflammation, smoking, occupational hazards, and industrial environmental pollution. Conclusions. Though etiology of sinonasal inverted papilloma remains controversial, the studies reviewed here indicate the importance of viral infection, cell cycle and angiogenic factors, environmental and occupational exposure, and chronic inflammation. Further studies on etiologic factors are required to ensure better clinical guidance and therapeutic targets.

Rinosporidiose equina: relato de caso no centro-oeste brasileiro

This is the first report of equine nasal rhinosporidiosis in Mineiros, Goiás, diagnosed through the histopathological analysis of granulomatous masses removed from a horse’s nostril of a 12-year-old male horse, through a surgical procedure. Microscopy revealed a papillomatous tissue lesion covered by moderately hyperplastic squamous epithelium, rich, loose fibrocollagenous tissues, well vascularized, accompanied by an accentuated inflammatory reaction constituted by a mixture of macrophages, lymphocytes, plasmocytes, and smaller number of neutrophils, and numerous round to oval structures with a unilamellar wall and a central nucleus surrounded by basophilic granular material, which were morphologically compatible with Rhinosporidium spp. sporangia.

Profiling of VEGF Receptors and Immune Checkpoints in Recurrent Respiratory Papillomatosis.

Recurrent respiratory papillomatosis (RRP) is caused by human papilloma virus (HPV) infection of the aerodigestive tract that significantly impacts quality-of-life including the ability to communicate and breathe. Treatment was traditionally limited to serial ablative procedures in the O.R. with possible local adjuvant therapy, but new systemic therapies, such as Vascular endothelial growth factor (VEGF) inhibitors, are showing significant promise. This study aims to determine whether rationale exists for combination therapeutic approaches using VEGF inhibitors and/or immune checkpoint blockade. Using fresh specimens from the O.R., we performed flow cytometry on papilloma, normal adjacent tissue, and blood. Papilloma and surrounding tissue were examined for expression of PD-L1, PD-L2, Galectin-9, VEGFR2, and VEGFR3. CD8+ and CD4+ T cells were assayed for expression of PD-1, TIGIT, LAG3, and TIM3. Our data shows that papilloma tissue exhibits significantly higher levels of PD-L1 and PD-L2 compared to adjacent tissue. Elevated levels of the VEGF receptor VEGFR3 were also observed in papilloma tissue. When examining T cells within the papilloma, elevated PD-1 and TIGIT expression was observed on CD8+ T cells, while levels of PD-1, TIGIT, and TIM3 were elevated on CD4+ T cells compared to PBMCs. Heterogenous marker expression was observed between individuals. Our analysis shows that RRP tissue shows elevated levels of multiple immune check point targets and VEGFR3, with varied patterns unique to each papilloma patient. Some of these immune checkpoint markers already have novel immunotherapies available or in development, providing molecular rationale to offer these systemic treatments to selected patients affected by RRP alongside VEGF inhibitors. Laryngoscope, 134:2819-2825, 2024.

Abstract 4012: EGFR inhibition in combination with PI3K/AKT/mTOR and/or Bcl-2 inhibition is a promising novel therapeutic approach for inverted sinonasal papillomas and associated sinonasal carcinomas

Abstract Introduction: Activating EGFR mutations are characteristic of inverted sinonasal papillomas (ISP) and ISP-associated sinonasal carcinomas (SNC). Using standard 2D culture models, our group has shown that targeting EGFR activity in ISP-associated SNC effectively inhibits cellular proliferation in vitro. Targeting EGFR in pre-clinical models that are more representative of in vivo patient responses is a critical next step. Methods: High-throughput drug screening was conducted using a 3D in vitro culture model for two ISP-associated SNC cell lines (SCCNC4 and UM-SCC-112) using a library of >800 curated small molecule inhibitors including inhibitors of EGFR, PI3K/AKT/mTOR (P/A/M), and Bcl-2. Viability was assessed and drug sensitivity scores were calculated. Dual combinations (inhibition of EGFR and P/A/M or Bcl-2) and triple combinations (inhibition of EGFR, P/A/M, and Bcl-2) for select drugs were tested for synergistic killing using the Chou-Talalay and Bliss methods. Treatment-associated transcriptomic changes were monitored using whole-transcriptome RNAseq. Finally, patient-derived organoids (PDOs) from primary sinonasal papilloma or carcinoma tissue were utilized ex vivo for short-term drug screening with EGFR, P/A/M, and/or Bcl-2 inhibitors at maximum human plasma concentration (Cmax) format. Results: Drug screening demonstrated that ISP-associated SNC cell lines are highly resistant to a diversity of standard and targeted therapeutic agents. Indeed, these cell lines demonstrated poorer responses than other highly resistant tumor types previously analyzed by our group, including triple-negative breast cancer and bladder cancer. Our data demonstrated that EGFR inhibition alone does not effectively kill ISP-associated SNC cells in 3D culture; however, dual and triple combinations of EGFR and P/A/M and/or Bcl-2 inhibitors showed synergistic killing. Finally, sinonasal papilloma and carcinoma PDOs showed partial responses to selected EGFR inhibitors in 3D ex vivo culture. Conclusions: While EGFR inhibitors effectively inhibit cellular proliferation in ISP-associated SNC cell lines in 2D culture, drug screening in 3D culture and using PDO material - more representative models of in vivo patient responses - show that EGFR inhibition alone is insufficient to completely eliminate these tumor cells, suggesting that EGFR inhibitors alone are unlikely to generate significant clinical responses for patients with ISP or associated SNCs. However, combining targeting of the P/A/M and/or Bcl-2 pathways revealed synergistic killing of ISP-associated SNC cell lines. Taken together, these data demonstrate that compensatory oncogenic and/or anti-apoptotic pathways must be targeted to potentiate EGFR inhibition for successful treatment of ISP and associated SNCs. Citation Format: Athena M. Apfel, Zhaoping Qin, Albert Liu, Matthew B. Soellner, Sofia D. Merajver, Aaron M. Udager, Nathan M. Merrill. EGFR inhibition in combination with PI3K/AKT/mTOR and/or Bcl-2 inhibition is a promising novel therapeutic approach for inverted sinonasal papillomas and associated sinonasal carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4012.

Toll-like receptor agonists, poly(I:C) and flagellin, lead to IL-36γ induction with divergent release kinetics and differentially alter autophagy in primary human keratinocytes

IL-36γ, a pro-inflammatory member of the IL-1 cytokine superfamily, can be induced and secreted by normal human foreskin keratinocytes (HFKs) in response to pathogenic stimuli, however, the mechanisms underlying the secretion are unknown. In this study, we demonstrate that stimulation with the TLR3 agonist, poly (I:C), led to a delayed secretion of IL-36γ compared to stimulation with the TLR5 agonist, flagellin, despite equal levels of the cytokine (p = 0.006). IL-36γ was shown to be released from HFKs in its inactive, uncleaved form, based on western blotting. Moreover, recombinant IL-36γ in its activated, cleaved form induced endogenous IL-36γ 10-fold (p = 0.004) and CXCL8 five-fold (p = 0.003) over baseline levels compared to unactivated full-length recombinant IL-36γ. The ratio of LC3b-II/LC3b-I was significantly higher in poly(I:C)-treated cells compared to flagellin-treated and unstimulated controls without a change in SQSTM1/p62 after 24 hours of stimulation (p = 0.043). Under fluorescence microscopy, poly(I:C) led to a two-fold increase at eight hours and four-fold increase at 24 hours in accumulated autophagosomes post-stimulation (p = 0.032). In contrast, autophagosomes were unchanged relative to baseline in response to flagellin. Bafilomycin A1 treatment enhanced poly(I:C)-mediated IL-36γ secretion (p = 0.044) while rapamycin led to a noticeable, but non-significant, increase in flagellin-mediated IL-36γ secretion, indicating that interrupting autophagic flux can alter IL-3γ grelease from HFKs. Finally, we show that, compared to clinically normal laryngeal tissue, there were significantly higher levels of LC3b-II in HPV-infected respiratory papilloma tissue, indicating a higher number of autophagosomes; a signature of disrupted autophagic flux.

Higher Number of EBI3 Cells in Mucosal Chronic Hyperplastic Candidiasis May Serve to Regulate IL-17-Producing Cells.

Previous research into the inflammatory cell infiltrate of chronic hyperplastic candidosis (CHC) determined that the immune response is primarily composed of T cells, the majority of which are T helper (CD4+) cells. This present investigation used immunohistochemistry to further delineate the inflammatory cell infiltrate in CHC. Cells profiled were those expressing IL-17A cytokine, EBI3 and IL-12A subunits of the IL-35 cytokine, and FoxP3+ cells. Squamous cell papilloma (with Candida infection) and oral lichen planus tissues served as comparative controls to understand the local immune responses to Candida infection. The results demonstrated that Candida-induced inflammation and immune regulation co-exist in the oral mucosa of CHC and that high prevalence of cells expressing the EBI3 cytokine subunit may play an important role in this regulation. This balance between inflammation and immune tolerance toward invading Candida in the oral mucosa may be critical in determining progress of infection.

083 Chemically-induced cutaneous neoplasms spontaneously regress in mice lacking autoimmune regulator

Cutaneous squamous cell carcinomas (cSCCs) frequently arise from precancerous lesions known as actinic keratoses (AKs). Factors that determine whether an AK will progress to a cSCC, remain stable, or regress to normal tissue are poorly defined. In AK and cSCC mouse models upregulation of autoimmune regulator (Aire) is associated with skin inflammation and tumor onset and the genetic lack of Aire attenuates the early stages of skin tumorigenesis. Here, to better assess the role of Aire in the onset and promotion of AK- and cSCC-like lesions, we subjected germline Aire null mice (Aire-/-; FVB/NJ or C57BL/6J backgrounds) to a classic two-step chemical carcinogenesis protocol [1x 7,12-dimethylbenz[a]anthracene (DMBA), 2x/wk 12-O-Tetradecanoylphorbol-13-acetate (TPA)], which resulted in papilloma formation in 92% of all mice (n=36/39). Aire deficiency resulted in reduced papilloma burden and smaller sized papillomas compared to Aire+/+ control mice. Unexpectedly, we observed that all papillomas that formed in 12 out of 22 Aire-/- mice rapidly and synchronously regressed despite continued tumor promotion with TPA. Histopathology indicated that regressed papilloma tissue from Aire-/- mice more closely resembled normal mouse skin architecture than non-regressed papilloma tissue from Aire-/- mice or from Aire+/+ papilloma tissue. After 15 weeks of TPA treatment, CD8+ cells consistent with cytotoxic T cell infiltrate were observed in the stroma surrounding the hyperproliferative epithelium in non-regressed papilloma tissue from Aire-/- mice or Aire+/+ mice. However, these cells were markedly absent from regressed papilloma tissue in Aire-/- mice, suggesting that an absence of functional Aire may improve the anti-tumoral immune response during the early stages of skin tumorigenesis. Altogether, this study indicates that Aire contributes in multiple ways to the development of skin neoplasms (onset, promotion, and regression) and suggests that the targeting of Aire function may provide a therapeutic benefit to patients at risk for skin cancers.

MACE RNA sequencing analysis of conjunctival squamous cell carcinoma and papilloma using formalin-fixed paraffin-embedded tumor tissue

Recent advances in the field of biomedical research allow for elucidation of the transcriptional signature of rare tumors such as conjunctival squamous cell carcinoma (SCC). In this study we compare its expression profile to conjunctival papilloma (Pap) and healthy conjunctival tissue (Ctrl) and develop a classification tool to differentiate these entities. Seven conjunctival SCC, seven Pap and ten Ctrl were formalin-fixed and paraffin-embedded (FFPE) and analyzed using Massive Analysis of cDNA Ends (MACE) RNA sequencing. Differentially expressed genes (DEG) and gene ontology (GO) clusters were explored and the abundance of involved cell types was quantified by xCell. Finally, a classification model was developed to distinguish SCC from Pap and Ctrl. Among the most prominent DEG in SCC a plethora of keratins were upregulated when compared to Pap and Ctrl. xCell analysis revealed an enrichment of immune cells, including activated dendritic cells and T-helper type 1 cells (Th1), in SCC when compared to Ctrl. The generated classification model could reliably discriminate between the three entities according to the expression pattern of 30 factors. This study provides a transcriptome-wide gene expression profile of rare conjunctival SCC. The analysis identifies distinct keratins, as well as dendritic and Th1 cells as important mediators in SCC. Finally, the provided gene expression classifier may become an aid to the conventional histological classification of conjunctival tumors in uncertain cases.

Presence of Tertiary Lymphoid Organ in Nasal Inverted Papilloma Is Correlated with Eosinophil Infiltration and Local Immunoglobulin Production

Introduction: Nasal inverted papilloma (NIP) is a benign tumour with multiple inflammatory cell infiltration. Tertiary lymphoid organs (TLOs) support local antibody production and play important roles in airway inflammation. However, the evidence of TLOs and local immunoglobulins in NIP has not been reported yet. We investigated the presence of TLOs and immunoglobulins in NIP tissues and their association with the clinical-pathological characteristics of NIPs. Methods: We analyzed the occurrence and composition of TLOs and local immunoglobulins by immunohistochemistry and evaluated the lymph organogenesis associated genes and cytokines by quantitative qPCR and Luminex assays, respectively, in papilloma tissues from 84 NIP cases. Results: TLOs were present in 54% (45/84) of the NIP patients but not in control subjects. TLOs were composed of T cells, B cells, follicular dendritic cells, macrophages, and natural killer cells. Compared to NIP tissues without TLOs, tissues with TLOs showed significantly higher eosinophil infiltration levels (3.5-fold), elevation of lymphorganogenic genes (CXCL12, CXCL13, CCL20, CCL21, CD21L, and lymphotoxin alpha and beta), and increased Th17 (IL-21, IL-22, and GM-CSF) and Th2 (IL-5 and IL-13) cytokine production. Moreover, NIP with TLOs demonstrated a higher number of follicular T helper cells and immunoglobulin-producing plasma cells (CD138+ IgA+, CD138+ IgM+, CD138+ IgE+, and CD138+ IgG+) than those without TLOs, and these antibody-producing cells were positively correlated with the eosinophil number. Conclusion: The high frequency of TLOs and excess local immunoglobulin production are associated with an eosinophilic and Th2 skew microenvironment in the NIP mucosa, which would contribute to an important immunopathogenic response during NIP pathogenesis.

Promotion of BZW2 by LINC00174 through miR-4500 inhibition enhances proliferation and apoptosis evasion in laryngeal papilloma

BackgroundWe aimed to explore the roles of basic leucine zipper and W2 domains (BZW) 2 in the human papillomavirus-infected laryngeal papillomatosis.MethodsIn the present study, BZW 2 knockdown and overexpressed cell lines were constructed. CCK-8 and colony formation assays were used to determine cell proliferation. Caspase-3 activity and nucleosomes fragmentation assays were used to determine cell apoptosis. qRT-PCR and Western blot were employed to evaluate the mRNA and protein levels of target genes, respectively. Luciferase and biotin-coupled miRNA pulldown assays were used to examine the interactions between mRNA and mRNA.ResultsWe observed the levels of BZW2 were up-regulated in the laryngeal papilloma (LP) tissues as compared with adjacent tissues. The knockdown of BZW2 significantly inhibited cell proliferation and promoted cell apoptosis in the LP cells. Additionally, we identified the expressions of BZW2 negatively regulated by miR-4500. Luciferase and biotin-coupled miRNA pulldown assays demonstrated that LINC00174 competed with the BZW2 for binding with miR-4500. Moreover, the results showed that LINC00174/miR-4500/BZW2 axis regulated cell proliferation and apoptosis.ConclusionOur results demonstrated that the regulation of LINC00174/miR-4500/BZW2 axis might be used as an effective strategy for treatment of human papillomavirus-infected laryngeal papillomatosis.

The possible role of Stomoxys calcitrans in equine sarcoid transmission

The association between bovine papillomavirus (BPV) and equine sarcoids is well established, but it is unclear how the virus spreads. Although evidence in support of viral spread through direct animal contact exists, this does not explain sarcoid development in isolated equids. BPV DNA has been detected in flies, which could indicate that these insects serve as a vector. This study aimed to investigate whether BPV-negative stable flies (Stomoxys calcitrans) become positive for BPV DNA after exposure to equine sarcoid or bovine papilloma tissue under experimental conditions and, if so, for how long. A total of 420 stable flies were caught alive and exposed to BPV positive equine sarcoid or bovine papilloma tissue. During the following week, dead flies were collected daily and BPV loads were determined by quantitative PCR. There was a significant rise in BPV load after tissue exposure both in sarcoid and papilloma exposed flies, but the viral load was higher and remained high for a longer time after exposure to papilloma tissue compared to sarcoid tissue. Within days, viral loads decreased again and became indifferent from loads before exposure. The results of these experiments indicate that BPV transmission by S. calcitrans seems possible and is more likely to occur after contact with bovine papillomas than with equine sarcoids. Transmission seems only possible shortly after tissue exposure. Further research could include experimental induction of sarcoids with BPV positive stable flies, or a repeat of the experiment with micro-dissection prior to PCR.

Analyzing esophageal squamous cell papillomas for the presence of human papilloma virüs.

Human Papilloma Virus (HPV) infection can be a predisposing condition for the development of squamous cell papilloma (SCP) of the esophagus, which can progress to dysplasia and to carcinoma as a result of chronic infection. The aim of the present study was to search for the presence of HPV in the esophageal SCP, and to genotype the detected HPV. Data from patients with definite diagnosis of SCP of the esophagus were identified from pathology records for two years period at different Hospitals. Slides from each patient were reviewed and samples with satisfactory papilloma tissues were submitted to molecular analysis. DNA has been isolated. DNA sequencing has been performed for genotyping HPV for all types. Our study group consisted of 21 women and 17 men (a total of 38 patients), mean age was 41 years (range 17-67 years). Most of the papillomas were located at mid-esophagus (68%). Eight out of 38 patients (21%) had associated erosive esophagitis, and fourteen patients (36.8%) had Helicobacter Pylori (H. pylori). Of the 38 SCP analyzed, seven (19%) were positive for HPV DNA. Three of them were of genotype 6, whereas four were of genotype 16, 18, 31, 81 that are known as highly oncogenic. There were no correlations between the presence of HPV and the patient's age, the presence of reflux esophagitis or H. pylori, smoking habit and the location of the papillomas. The presence of high-risk type HPV in esophageal SCP may implicate a role of the virus in the pathogenesis of the esophageal tumor.

Poly(I:C) induces controlled release of IL-36γ from keratinocytes in the absence of cell death.

The epithelium is part of an integrated immune system where cytokines, toll-like receptors and their ligands, and extracellular vesicles play a crucial role in initiating an innate immune response. IL-36γ is a pro-inflammatory member of the IL-1 family that is mainly expressed by epithelial cells, but regulation of its expression and release are only beginning to be understood. Previous studies reported that IL-36γ is abundant in recurrent respiratory papillomatosis, a rare but devastating disease caused by human papillomaviruses (HPV) types 6 and 11, in which papillomas recurrently grow in and block the airway. Despite the overexpression of IL-36γ, papilloma tissues show no evidence of inflammation, possibly due to suppression of its release by HPVs. We have used primary human foreskin keratinocytes as a model to study IL-36γ regulation in normal epithelial cells. Low doses of poly(I:C) mediate expression and release of IL-36γ without inducing the cell death reported by those using high doses. PKR, an enzyme required for inflammasome activation, does not contribute to controlled release of IL36γ. The keratinocytes secrete IL-36γ in two forms, soluble and in extracellular vesicles. We conclude that there are two separately regulated pathways for the controlled secretion of IL-36γ from keratinocytes, which could contribute to the modulation of both local and systemic immune responses to viruses and other pathogens.

The effect of metallothionein 2A core promoter region single-nucleotide polymorphism on accumulation of toxic metals in sinonasal inverted papilloma tissues

Metallothioneins (MTs) are intracellular thiol-rich heavy metal-binding proteins which join trace metal ions protecting cells against heavy metal toxicity and regulate metal distribution and donation to various enzymes and transcription factors. The goal of this study was to identify the −5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene, and to investigate its effect on allele-specific gene expression and Cd, Zn, Cu and Ni content in sinonasal inverted papilloma tissue (IP), with non-cancerous sinonasal mucosa (NCM) as a control. The MT2A promoter region −5 A/G SNP was identified by restriction fragment length polymorphism using 117 IP and 132 NCM. MT2A gene analysis was performed by quantitative real-time PCR. Metal levels were analyzed by flame atomic absorption spectrometry. The frequency of A allele carriage was 99.2% and 100% in IP and NCM, respectively. The G allele carriage was detected in 23.9% of IP and in 12.1% of the NCM samples. As a result, a significant association of −5 A/G SNP in MT2A gene with mRNA expression in both groups was determined. A significant association was identified between the −5 A/G SNP in the MT2A gene with mRNA expression in both groups. A highly significant association was detected between the rs28366003 genotype and Cd and Zn content in IP. Furthermore, significant differences were identified between A/A and A/G genotype with regard to the type of metal contaminant. The Spearman rank correlation results showed the MT2A gene expression and both Cd and Cu levels were negatively correlated. The results obtained in this study suggest that the −5 A/G SNP in the MT2A gene may have an effect on allele-specific gene expression and toxic metal accumulation in sinonasal inverted papilloma.

Morphological characteristics of conjunctival squamous papillomas in relation to human papillomavirus infection

ObjectiveTo determine the prevalence of a broad spectrum of human papillomavirus (HPV) types in conjunctival papillomas and a possible difference in clinical and histopathological presentation of HPV-positive and HPV-negative papillomas.MethodsFormalin-fixed,...

Genetic dysregulation in recurrent respiratory papillomatosis

Recurrent respiratory papillomatosis (RRP) is a devastating disease, caused by infection of the upper aerodigestive tract with human papillomavirus types 6 and 11. There is no cure for RRP, and surgical removal is the mainstay of treatment. The purpose of this project was to compare genes of cell cycle, apoptosis, and inflammatory cytokines in laryngeal papilloma versus normal tissue for a better understanding of the molecular mechanisms of the disease to discover novel therapies. Basic science research study. Papilloma tissue was obtained from patients requiring surgical debridement. For comparison, normal mucosa was obtained from the excised uvula of patients undergoing uvulopalatopharyngoplasty. Total RNA was extracted from both groups and then probed using customized reverse transcriptase real time polymerase chain reaction gene arrays. The custom arrays examine expression of 84 separate genes within the cell cycle, apoptosis, and inflammatory cytokine pathways. Our findings based on 11 papilloma samples run in comparison to normal mucosa shows that the MCL-1 gene of the apoptosis pathway is significantly downregulated. cytokine genes IL1-A, IL-8, IL-18, and IL-31 are also significantly dysregulated. Genes of cell cycle and apoptosis are generally upregulated and downregulated, respectively, as expected in papilloma tissue, with MCL-1 achieving significance when compared to normal tissue. The finding of particular interest is that inflammatory cytokine genes were significantly downregulated, including IL1-A, IL-18, and IL-31. This finding may explain why patients infected with the virus are unable to mediate a T-cell immune clearance of their disease.

Papillomavirus-associated Cutaneous Papillomas in a Population of Wild Spotted Hyenas (Crocuta crocuta)

Beginning in 1997 Michigan State University Mara Hyena Project investigators observed waxing and waning progression of oral and genital masses during long-term behavioral observations of a population of wild spotted hyenas (Crocuta crocuta) from the Masai Mara Game Reserve, Kenya. From 1999-2000, we darted adult spotted hyenas to obtain routine physiologic and hematologic data and collected small, raised, lobulated, pigmented masses from the oral or genital areas of eight animals. Microscopically, masses consisted of variably thickened epidermis with thick elongate rete pegs, prominent stratum spinosum, and few koilocytes, consistent with papillomavirus-induced lesions. Immunohistochemistry on formalin-fixed, paraffin-embedded papilloma tissue revealed positive intranuclear labeling for papillomavirus antigen in the superficial stratum granulosum and in sloughing keratin layers of multiple samples. Polymerase chain reaction on DNA extracts from tumor tissue amplified a papillomavirus-specific 418 base pair amplicon in the E1 ORF. Basic Local Alignment Search Tool analysis of the sequenced amplicon suggests a novel hyaenid papillomavirus. Confirmatory complete genomic sequencing was performed later by the Rega Institute in Belgium. To our knowledge, this is the first report of a papillomavirus in a Hyaenidae species. Spotted hyena social behavior might facilitate oral-genital transmission of papillomavirus in this population.

Lung Squamous Cell Carcinoma Arising in a Patient with Adult-onset Recurrent Respiratory Papillomatosis

A 75-year-old male was admitted to our hospital in December 2011 with a mass in the right upper pulmonary lobe. He was incidentally diagnosed as having tracheal papillomas 10 years ago. Bronchoscopy revealed multiple polypoid papillomas in the dorsal lesion of the trachea. Polymerase chain reaction amplification detected human papillomavirus type 11 DNA in the papilloma tissues. A computed tomography scan demonstrated the occlusion of the right superior segment bronchus with distal consolidation. Furthermore, F-18 fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography showed intense tracer uptake in the right superior segment of the lung. He underwent a right upper lobectomy. The tumor was seen as a rounded nodule, ≈ 2 cm in diameter. Histological examination of the tumor revealed squamous papilloma with papillary and solid architecture surrounded by accumulation of acute inflammatory cells. Furthermore, in a part of the tumor, squamous cell carcinoma was also present. The lymph nodes were free of tumor. After the surgery, he continued to undergo endoscopic microwave resection. Recurrent respiratory papillomatosis is a rare disease that can cause life-threatening airway compromise and malignant transformation. The present case indicates that F-18 fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography is indispensable for early detection of lung cancer arising in a patient with recurrent respiratory papillomatosis.