Department of Otolaryngology, University of Rochester Medical Center, Rochester, New York.
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York.
JAMA Otolaryngol Head Neck Surg. 2018 Mar 1;144(3):231-237. doi: 10.1001/jamaoto.2017.2916.
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.
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.
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.
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.
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.
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.
人类乳头瘤病毒（HPV）相关口咽鳞状细胞癌的发病机制目前是阐明的重要课题。潜伏 HPV 感染在健康成人的扁桃体组织中的存在可能为该过程的一个组成部分提供解释，并有助于理解 HPV 相关口咽鳞状细胞癌的致癌作用。
确定口咽 HPV 的流行率，并确定病毒与扁桃体组织隐窝生物膜之间的空间关系。
设计、设置和参与者：这是一项回顾性、横断面研究，使用了 2012 年至 2015 年期间在一所大学医院因非肿瘤性扁桃体切除术而存档的扁桃体组织的福尔马林固定石蜡包埋样本进行。样本包括 102 名年龄在 20 至 39 岁之间的成年人的无肿瘤扁桃体组织的定量聚合酶链反应检测到高危亚型 16 和 18。通过聚合酶链反应评估 HPV 状态，并用原位杂交分析定位病毒衣壳蛋白。然后用伴刀豆球蛋白 A 对这些样本进行染色以确定生物膜的存在和形态。然后用二脒基苯基吲哚 (DAPI) 对这些样本进行染色以观察病毒与细胞核的位置关系。然后将这些数据组装起来进行综合分析，以确定 HPV 在扁桃体隐窝生物膜中的位置。
在收集数据之前确定了结果测量，包括在 otherwise healthy adults 的扁桃体组织中高危 HPV 类型 16 和 18 的流行率，以及用免疫组织化学法在扁桃体隐窝生物膜中检测到 HPV。
在 102 名 otherwise healthy adults（55 名[53.9%]为女性；年龄 20-39 岁）中，扁桃体 HPV 的总体流行率为 4.9%（n=5）；高危型 16 或 18 为 3.9%（n=4）。在该样本人群中，原位杂交将 HPV 病毒与扁桃体隐窝的生物膜进行了共定位。
生物膜存在于相当一部分扁桃体组织的扁桃体隐窝中，可以反复识别。已经证明 HPV 与隐窝生物膜共定位。这对口咽 HPV 流行率的确定具有重要意义。它也可能在 HPV 相关口咽癌的发病机制中起作用。
