The purpose of this study was to determine whether changes in human papillomavirus (HPV) DNA prevalence in oral rinses and/or HPV-specific antibody levels in the sera of patients with oral/oropharyngeal cancer have prognostic significance. One hundred and forty-two patients with oral/oropharyngeal tumors were enrolled. The presence of HPV DNA was assayed in tumor tissue and oral rinses and HPV-specific antibodies were assessed in the sera. Oral rinses were collected before treatment and one year after the treatment. Sera were drawn before treatment, one month, and one year after the end of the treatment. Altogether, 59.2% of tumors were HPV positive. The presence of HPV DNA in the tumors correlated with HPV DNA positivity in oral rinses and with HPV-specific antibodies in the sera. Out of 66 patients with HPV-positive oral rinses at enrolment, 84.8% became negative at one-year follow-up, while most patients remained seropositive for HPV-specific antigens. However, the mean titers of HPV16 E6 and/or E7 antibodies at follow-up were significantly lower. Of 16 patients with recurrences at followup (alive on second sampling), six were positive at enrolment for HPV16 E6 and/or E7 antibodies. In five of these, no decrease in antibody levels was observed. Titers of antibodies specific for HPV16 capsid antigens did not change during the follow-up. Our data suggest that the detection of antibodies specific for the HPV 16 E6 and E7 oncoproteins may serve not only as a marker of HPV etiology, but also as a marker of recurrence and a prognostic indicator in patients with HPV-positive tumors.Squamous cell carcinoma of the head and neck (SCCHN) accounts annually for 7% of all new cancer cases worldwide. 1 SCCHN presents as a single clinical and pathological entity, yet studies conducted over the past 15 years have revealed that it has two distinct etiologies. Most SCCHN cases, particularly those located in the oral cavity, larynx, and hypopharynx, are apparently independent of viral infection, and often associated with tobacco and alcohol use. However, 26% of all SCCHN cases, and over 70% of those found in the tonsillar area and oropharynx, are associated with the presence and expression of mucosal high-risk (oncogenic) human papillomavirus (HR HPV) genomes. [2][3][4] HR HPVs cause cervical cancer and other neoplasms of the anogenital skin and mucosae. 5,6 The most common genital mucosal HR HPV type 16, is found in 95% of HPVpositive SCCHN lesions. 7,8 Although patients with HR HPVassociated SCCNH often present with more advanced disease, we and others have shown that these patients have a remarkably better prognosis and five-year survival rate. 2,9-11 The prognostic advantage of patients with SCCHN etiologically linked to HR HPV could potentially lead to modified treatment regimens. Therefore, noninvasive or less invasive sampling procedures to identify patients with HPV-positive tumors were evaluated in several studies including ours. 12-16It has been demonstrated that the presence of HPV DNA in oral rinses or saliva as well...

The association of high-risk human papillomaviruses (HR HPVs) with tonsillar cancer (TC) has been documented. Because patients with HPV-associated tumors show better survival rates, modification of their treatment regimen is being considered. It is therefore crucial to find markers for the identification of patients whose tumors are linked to viral infection. A cohort of 109 patients with primary TC was screened for HPV DNA presence in the tumor tissues and HPV-specific antibodies in sera. Data regarding risk factors and clinical parameters were collected. Forty-five specimens were analyzed for the expression of viral E6 and E2-region mRNA, and the p16 and p53 protein expression status was assessed by immunohistochemistry. The overall prevalence of HPV DNA in TC tissues was 65.1%. Ninety-three percent of HR HPV DNA-positive samples expressed E6*I mRNA. E2-region mRNA expression was detected in 36% of positive samples, which implies that the virus is integrated in 64% of HPV DNA/RNA-positive tumors. p16 overexpression and the presence of antibodies specific to HPV16 E6/E7 oncoproteins correlated well with HPV DNA and RNA presence. The disease-specific survival rate of patients with HPV DNApositive tumors was significantly higher than that of HPV DNA-negative patients. In addition to providing further evidence of the involvement of HPV infection in the etiopathogenesis of a proportion of TC cases, our study demonstrates that p16 immunostaining and anti-E6/E7 antibodies as surrogate markers of HPV involvement represent specific, sensitive and clinically accessible assays for the identification of TC patients who have a considerably better prognosis.Recent studies have shown that 20-25% of head and neck squamous cell cancers (HNSCCs) appear to be causally linked with high-risk human papillomavirus (HR HPV) infection.

Prognostic factors are important for treatment decisions as they help adapt the therapy on a case-to-case basis. Nodal status, number of positive nodes, and presence of extracapsular spread are considered to be the important prognostic factors in head and neck cancer. Some studies suggest that human papillomavirus (HPV) status also influences the outcome of the treatment. This influence can be explained by the variation in tendency to develop regional metastases and by variation in the type of neck node involvement. The study objectives were to compare patients with HPV positive and HPV-negative tumors for survival and prevalence and type of regional metastasis, to identify prognostic factors and to test whether HPV presence is an independent factor of survival. The study included 81 patients treated by surgery including neck dissection for oral or oropharyngeal squamous cell cancer. A computerized medical report was completed for each patient. Analysis of the tumor specimen for the HPV DNA presence was done on paraffin-fixed tissue. HPV DNA detection and typing were performed by PCR with GP5+/GP6+BIO primers and reverse line blot hybridization. Overall, 64% (52/81) of tumors were HPV positive with 80% in the tonsillar site. HPV-positive patients had significantly better both overall (73 vs. 35%) (P=0.0112) and disease-specific (79 vs. 45%) (P=0.0015) survival rates than HPV-negative patients. No significant differences were found in the pN classification, in the number of positive nodes and the presence of extracapsular spread in the involved nodes between HPV positive and HPV-negative tumors. Multivariate analysis showed that significant prognostic factors of survival were the presence of HPV in the tumor, extracapsular spread and tumor size. HPV was the most significant prognostic factor in the studied group of patients with oropharyngeal tumors (HR=0.27, 95%CI 0.12-0.61) and possibly should be considered in treatment decisions.

The association between human papillomavirus (HPV) infection and the development of head and neck cancer has been documented recently. In this study on 86 head and neck cancer patients and 124 controls, data regarding demographics, behavioral risk factors, and risks related to HPV exposure were collected. HPV detection was carried out using polymerase chain reaction in the tumors and in oral exfoliated cells, and HPV typing by a reverse line blot assay specific for 37 HPV types. Sera were tested by an enzyme-linked immunosorbent assay specific for HPV proteins. Head and neck cancer cases report significantly more oral-anal contact (P = 0.02) and tobacco and alcohol use than controls (P = 0.001; P = 0.02, respectively). High-risk HPV DNA was detected in 43% of oral washings of cases and 4% of controls (P < 0.0001). The association between the presence of high-risk HPV DNA in oral exfoliated cells and in tumor tissues was statistically significant (adjusted P < 0.0001). The prevalence of HPV-specific antibodies was significantly higher in cases than in controls (adjusted P < 0.0001). These results provide epidemiological and immunological evidence for HR HPV as a strong risk factor (OR = 44.3, P < 0.0001) for head and neck cancer, even after controlling for age, tobacco and alcohol use. The detection of high-risk HPV DNA in oral exfoliated cells and HPV-specific antibodies in serum can be considered as clinically relevant surrogate markers for the presence of a HPV-associated head and neck cancer, with a high sensitivity (83%) and specificity (88%).

As more total laryngectomies (TLE) are nowadays performed as salvage procedures, the rate of postoperative complications increases. The primary aim was to report the rates of postoperative local complications for total laryngectomy in patients with previous radiotherapy or chemoradiotherapy (RT/CRT) in comparison with primary TLE data. We attempted to identify patient-and tumor-related factors predictive of postoperative pharyngocutaneous Wstula (PCF) formation. The secondary aims were to analyze the survival rate in relation to postoperative complications and to study prognostic factors of survival in TLE patients. A retrospective study was conducted in 208 patients. Logistic regression was used to determine the most signiWcant risk factors for Wstula formation. Survival was analyzed by the Kaplan-Meier method, log-rank test, and Cox multivariate regression. PCF developed in 20.7% of cases. In the group of patients with previous RT/CRT, the Wstula rate was signiWcantly higher (34%). In multivariate analysis, signiWcant risk factors for Wstula formation were previous radiotherapy or chemoradiotherapy (p = 0.02), higher N classiWcation (p = 0.03), and procedure performed by a less experienced surgeon (p = 0.003). The survival and recurrence rates were not inXuenced by PCF formation. The overall survival rates were lower in patients with previous RT/CRT and in patients with lymph node involvement.

Background. Human papillomaviruses (HPVs) have been proved as one of the etiological factors of oropharyngeal squamous cell carcinoma (OPSCC). Patients with tumors of viral etiology have a lower recurrence rate and better prognosis. OPSCC is linked to an alteration in the immune system. Only a limited number of studies have correlated both the immunological parameters and HPV status with patient prognosis. The aim of this study was to determine whether HPV infection and the immunological status influence patient prognosis individually or in concurrence. Material and Methods. Sixty patients with oral and oropharyngeal carcinomas were enrolled. They were divided into HPV-positive and HPV-negative groups based on the expression of HPV 16 E6 mRNA. Basic lymphocyte subpopulations were determined in the peripheral blood by means of flow cytometry. Results. Significantly better disease-specific survival (DSS) was observed in patients with HPV-positive tumors. Nodal status, tumor grade, recurrence, and CD8+/Tregs ratio were identified as factors influencing DSS. A higher level of Tregs and a lower ratio of CD8/Tregs influenced overall survival (OS) independently of HPV status and age. Patients with HPV-positive tumors and high levels of Tregs survived significantly better than patients from the other groups. Conclusion. Better survival is associated with HPV positivity and elevated Tregs levels. Our data suggest that HPV infection and Tregs do not influence patient prognosis in concurrence.

The difference in the prevalence of HPV DNA positive tumours between cases of oral cavity and oropharyngeal carcinoma exposed and not exposed to tobacco or alcohol support the theory that HPV DNA positive tumours form an aetiologically distinct subgroup of head and neck tumours.

Integration, which leads to the disruption of the circular HPV genome, is considered as a critical, albeit not obligatory, step in carcinogenic progression. Although cervical carcinomas with extrachromosomal HPV plasmid genomes have been described, the virus is integrated in 70% of HPV16-positive cervical tumours. Limited information is available about HPV integration in head and neck tumours (HNC). In this study, we have characterised the physical status of HPV in a set of tonsillar tumour samples using different methods-the mapping of E2 integration breakpoint at the mRNA level, the 3' RACE based Amplification of Papillomavirus Oncogene Transcripts (APOT) assay and Southern blot. Furthermore, the impact of HPV integration on patients' prognosis has been evaluated in a larger set of 186 patients with head and neck cancer. Based on the analysis of E2 mRNA, HPV was integrated in the host genome in 43% of the HPV-positive samples. Extrachromosomal or mixed form was present in 57%. In fresh frozen samples, the APOT and E2 mapping results were in agreement. The results were confirmed using Southern blotting. Furthermore, the type and exact site of integration were determined. The survival analysis of 186 patients revealed HPV positivity, tumour size and lymph node positivity as factors that influence disease specific survival. However, no statistically significant difference was found in disease specific survival between patients with HPV-positive integrated vs. extrachromosomal/mixed forms of the virus.Head and neck cancer (HNC) is the seventh most common cancer in men and the 13 th most common cancer in women worldwide and its incidence is on the rise in some anatomical sites 1 . HNC has at least two different etiologies. Most HNC cases are associated with smoking and alcohol consumption, which induce mutations in important pathways 2,3 . However, 26% of all HNC and up to 50% of oropharyngeal tumours are associated with the presence and expression of mucosal high-risk human papillomaviruses (Alfa HR-HPV; HR-HPVs) [4][5][6] . Mucosal HRHPVs also cause cervical cancer and other neoplasms of anogenital skin and mucosae. The most common mucosal HR-HPV is HPV16, which is found in almost 90% of HPV-positive HNC, other common types being HPV18, HPV33 and HPV52 6,7 . Patients with HPV-associated HNC have a better prognosis, overall survival, and response to treatment 4 . The mechanisms of Alfa HR-HPV-induced carcinogenesis, regardless of the anatomical site, rely on the functions of viral oncoproteins, E6 and E7, as they inactivate tumour