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Original Investigation
March 2018

Prevalence of High-Risk Human Papillomavirus in Tonsil Tissue in Healthy Adults and Colocalization in Biofilm of Tonsillar Crypts

Author Affiliations
  • 1Department of Otolaryngology, University of Rochester Medical Center, Rochester, New York
  • 2Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York
  • 3Department of Pathology, University of Rochester Medical Center, Rochester, New York
  • 4University of Rochester, Rochester, New York
JAMA Otolaryngol Head Neck Surg. 2018;144(3):231-237. doi:10.1001/jamaoto.2017.2916
Key Points

Question  What is the prevalence of high-risk human papillomavirus (HPV) in otherwise healthy adult tonsil tissue, and does HPV colocalize in biofilm of tonsillar crypts?

Findings  In this retrospective, cross-sectional study, prevalence of HPV in tonsils of otherwise healthy adults is consistent with previously published studies. In our sample population, in situ hybridization colocalized HPV virus to the biofilm of the tonsillar crypts and demonstrated extranuclear presence of viral capsid.

Meaning  Colocalization of HPV in bacterial biofilm of human tonsil tissue has considerable implications with respect to the determination of HPV prevalence rates in the oropharynx and may also play a role in the pathogenesis of HPV-related oropharyngeal carcinoma.

Abstract

Importance  The pathogenesis of human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma is currently an important topic of elucidation. The presence of latent HPV infection in tonsil tissue of healthy adults may provide an explanation for a component of this process and contribute to the understanding of HPV-associated squamous cell carcinoma oncogenesis of the oropharynx.

Objective  To determine the prevalence of oropharyngeal HPV and to determine the spatial relationship between the virus and crypt biofilm in tonsil tissue.

Design, Setting, and Participants  A retrospective, cross-sectional study was carried out using samples obtained from tonsils that were archived at a university hospital following elective nononcologic tonsillectomy from 2012 to 2015. Samples consisted of formalin-fixed paraffin embedded samples of tumor-free tonsil tissue from 102 adults between the ages of 20 and 39 years.

Exposures  Human papillomavirus status was assessed by polymerase chain reaction, and high-risk subtypes 16 and 18 were assessed with quantitative polymerase chain reaction assay. Samples that demonstrated presence of HPV were then analyzed by in situ hybridization to localize the viral capsid protein. These samples were then stained with concanavalin A to establish biofilm presence and morphology. These samples were also stained with diamidino-phenylindole (DAPI) to visualize location of the virus in relation to cell nuclei. These data were then assembled for aggregate analysis to colocalize HPV in the biofilm of the tonsillar crypts.

Main Outcomes and Measures  Outcome measurements were determined prior to data collection and include prevalence of high-risk HPV types 16 and 18 in tonsil tissue of otherwise healthy adults, as well as demonstration with immunohistochemistry of HPV in tonsillar crypt biofilm.

Results  In 102 otherwise healthy adults (55 [53.9%] female; age range, 20-39 years), the overall prevalence of HPV in tonsils was 4.9% (n = 5); and high-risk type 16 or 18, 3.9% (n = 4). In this sample population, in situ hybridization colocalized HPV virus to the biofilm of the tonsillar crypts.

Conclusions and Relevance  Biofilm is present in the tonsillar crypts in a considerable proportion of tonsil tissues and may be reproducibly identified. Human papillomavirus is demonstrated to colocalize to the crypt biofilm. This has important implications with respect to the determination of HPV prevalence rates in the oropharynx. It may also play a role in the pathogenesis of HPV-related oropharyngeal carcinoma.

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