Presence of DNA of Human Papillomavirus 16 but No Other Types in Tumor-Free Tonsillar Tissue

Abstract

According to PCR, the prevalences of human papillomavirus (HPV) DNA were 6.3% (13 of 206) in tonsillitis or hypertrophic tonsillar tissues and 0.6% (1 of 174) in exfoliated cells from normal tonsils. HPV-16 was the only type detected in tonsillar tissues, but it did not appear to lead to L1 antibody production.Of the squamous cell carcinomas of the head and neck, human papillomavirus (HPV) DNA has been repeatedly reported in tonsillar carcinoma, suggesting a role for HPV in tonsillar carcinogenesis (5,9,10,13,22). The reason for the strong association of HPV with tonsils remains unclear. Previous data on the presence of HPV DNA in tumor-free tonsils are based on small series of experiments (24). A significantly increased risk for squamous cell carcinomas of the head and neck is associated with HPV-16 seropositivity (11), but serum antibodies against HPV-16 proteins have been detected in healthy individuals as well (6). Our aim was to study HPV in tumor-free tonsils and to correlate these findings with the seroprevalence of HPV antibodies.We enrolled 212 patients operated on in 2001 and 2002 because of tonsillitis (n ϭ 135) or tonsillar hypertrophy (n ϭ 77) at Helsinki University Central Hospital (Table 1). After tonsillectomy, a piece of tissue was immediately frozen at Ϫ70°C. A 100-m cryo-section per tonsil per patient was subjected to DNA extraction. Five-micrometer sections from both ends of the tissue were toluidine blue stained to verify that the tissue section consisted of at least 30% epithelial cells. After every tenth sample, only optimal cutting temperature compound without tonsillar tissue was cryo-sectioned as a negative control for the DNA extraction; none of these sections showed positive ␤-globin or HPV by PCR. Serum samples were drawn on the day of surgery.Tonsillar exfoliated cells from 189 control subjects (age and sex matched to patients) with normal tonsils were collected bilaterally from tonsillar surfaces with a Cytobrush Plus Cell Collector (Medscand Medical). The brushed samples were stored in phosphate-buffered saline at Ϫ70°C. Serum samples were collected on the same day.DNA was extracted from tonsillar tissue with the DNA mini kit (QIAGEN) and from exfoliated cells with the MagNA Pure LC DNA isolation kit I (Roche). In order to assess the quality of the DNA submitted to HPV PCR, we applied ␤-globin PCR (15) and found that 97% (206 of 212) of DNA samples from tonsillar tissue and 92% (174 of 189) of DNA samples from tonsillar exfoliated cells were positive for ␤-globin.HPV DNA was identified by nested PCR with consensus primers MY09/11 (14) and GP5ϩ/6ϩ (3). The genotype was determined by direct sequencing of the approximately 150-bp amplicons with an ABI PRISM Dye Terminator and analyzed on an ABI 377 DNA sequencer (Applied Biosystems) and on the basis of Ͼ90% homology with HPV sequences deposited in GenBank with BLAST software.Serum antibodies against HPV-16 L1, E6, and E7 were analyzed by a newly developed method. The HPV proteins were expressed as fusion proteins with N-terminal glut...

Bioportfolio: Lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue

“…Biopsies of skin (n ϭ 54) were obtained from patients with B19-unrelated dermatological lesions and from healthy hospital or laboratory staff (range, 1913-1991; mean, 1951 Ϯ 19). Biopsies of tonsillar tissue (n ϭ 220) were obtained during tonsillectomy from patients (range, 1929-2000; mean, 1979 Ϯ 15) with tonsillitis or tonsillar hypertrophy (44).…”

Human Papillomavirus and Related Diseases - From Bench to Bedside - A Clinical Perspective

“…They found 9 of 106 tonsils to be HPV-positive, with six cases having HPV-16, two cases having HPV-11 and one case having an untyped HPV (Mammas et al, 2006). Another study reported HPV-16 infection in 13 of 206 tonsils (6.3%) with 11 cases from children or young adults aged less than 25 years (Chen et al, 2005). These findings differed from those of several other studies in which low prevalence or no prevalence of oncogenic and non-oncogenic HPV was identified in the specimens studied (Brandsma & Abramson, 1989;Klingenberg et al, 2010;Klussmann et al, 2001;Niedobitek et al, 1990;Ribeiro et al, 2006;Sisk et al, 2006;Snijders et al, 1992).…”

Les cancers des voies aérodigestives supérieures associés aux papillomavirus

“…[12,13]. Cependant, d'autre modes de transmission sont suspectés, comme le suggère la détection (PCR) de HPV oncogènes dans des pièces d'amygdalectomie chez les enfants [14,15].…”