HPV-positive Cases Research Articles

Pre-microRNA variants predict HPV16-positive tumors and survival in patients with squamous cell carcinoma of the oropharynx

To identify non-tumor biomarkers for prediction of tumor HPV status and prognosis of patients with squamous cell carcinoma of the oropharynx (SCCOP), we evaluated the association of single nucleotide polymorphisms (SNPs) in pre-miRNAs with HPV16 status and survival for SCCOP patients. We analyzed HPV16 status in tumor specimens and genotyped four SNPs in pre-miRNAs (hsa-mir-146a rs2910164 G>C, hsa-mir-149 rs2292832 G>T, hsa-mir-196a2 rs11614913 C>T, and hsa-mir-499 rs3746444 A>G) in 309 SCCOP patients. Unconditional logistic regression models were used for calculation of odds ratio (OR) and 95% confidence intervals (CIs), and Kaplan–Meier analysis and Cox proportional hazards regression were used to evaluate associations. We found that statistically significant associations with HPV16-positive SCCOP and survival were found for hsa-mir-146a rs2910164 and hsa-mir-196a2 rs11614913, while such similar associations were not observed for hsa-mir-149 rs2292832 and hsa-mir499 rs3746444. Compared with those with corresponding hsa-mir-146a CG/CC and has-mir-196a2 CC genotypes, the hsa-mir-146a GG and hsa-mir-196a2 CT/TT wild-type genotypes were significantly associated with HPV16-positive tumor status (adjusted OR, 2.4; 95% CI, 1.4–4.1 and adjusted OR, 2.1, 95% CI, 1.2–3.6), respectively. Patients having hsa-mir-146a rs2910164 GG and hsa-mir196a2 rs11614913 CT/TT genotypes had significantly better overall, disease-specific, and disease-free survival compared with those having the corresponding CG/CC and CC genotypes, respectively. Furthermore, these genotypes were significantly associated with reduced risk of overall death, death owing to disease, and recurrence after adjustment for important prognostic confounders including HPV status, smoking, and stage. Our findings indicate pre-miRNA polymorphisms may predict tumor HPV16-positive SCCOP cases and may be prognostic biomarkers for SCCOP.

Nodal status is not a prognostic factor in patients with HPV‐positive oral/oropharyngeal tumors

The primary aim is to compare the prognostic parameters in patients with HPV-positive and HPV-negative tumors. The secondary aim is to compare the patterns of treatment failure between these groups. Analysis of prognostic factors in a group of 170 patients. High-risk HPV DNA was detected in 98 cases, 72 tumors were HPV negative. Both the overall and disease-specific survival rates were better in HPV-positive patients. In patients with HPV-negative tumors, the prognostic factors in univariate analysis were pT and pN classification, tumor stage, number of positive nodes, and extracapsular spread. Stage, pT, higher pN, and number of nodes maintained statistical significance after adjustment. None of the studied prognostic factors was significant in the group of patients with HPV-positive tumors. There was a significant difference in the local--regional recurrence rates--37% in HPV-negative cases and 18% in HPV-positive cases. The characteristics of the extent of the disease in general and of regional lymph node metastasis in particular are probably much less important in the prediction of the outcome of HPV-positive than of HPV-negative tumors. Improved survival of patients with HPV-positive tumors is due mostly to the difference in the local-regional failure rates.

Human Papillomavirus (HPV) and HPV 16–Variant Distribution in Vulvar Squamous Cell Carcinoma in Sweden

To investigate the human papillomavirus (HPV) and HPV type 16-variant distribution in a series of vulvar squamous cell carcinomas (VSCC) and to evaluate the impact of HPV and HPV 16-variant on prognosis. A series of 133 patients who had a diagnosis of VSCC (1983-2008) was selected for the study. Detection of 11 high-risk HPV types (16, 18, 31, 33, 39, 45, 51, 52, 56, 58, and 59) and 2 low-risk HPV types (6 and 11) was performed with real-time polymerase chain reaction. Samples positive for HPV 16 were further analyzed for variant determination of 7 positions in the E6 gene with polymerase chain reaction and pyrosequencing. Forty (30.8%) of 130 tumors were found to be HPV positive. Human papillomavirus type 16 was found in 31 cases, HPV 18 was found in 2 cases, HPV 33 was found in 5 cases, and HPV 56 and HPV 59 were found in one case each. All but one tumor harboring HPV 16 were of European linage, and the 3 most common variants were E-p (n = 13), E-G350 (n = 7), and E-G131 (n = 5). HPV positivity was associated with the basaloid tumor type and occurred in significantly younger patients. Overall and recurrence-free survival rates were better in HPV-positive cases, but after correction for age and tumor size, HPV status was no longer an independent and significant prognostic factor. The survival rates of the various HPV 16 variants were not significantly different, but there was a trend of worse outcome for the E-G131-variant group. Human papillomavirus positivity of 30.8% is similar to other reports on VSCC. To our knowledge, this first variant determination of HPV 16 in vulvar carcinoma in a Swedish cohort indicated that the variant E-G131 may have an increased oncogenic potential in patients with VSCC.

1040P - The Impact of Induction Chemotherapy on Cisplation Dose Intensity During Chemo-Radiotherapy (CTRT) in Oropharyngeal Squamous Cell Cancer (OPC)

ABSTRACT Background Induction (IC) with TPF (Taxotere, Cisplatin, 5-FU) has been associated with improved outcome in advanced head and neck squamous cell cancer patients (pts). Full dose of cisplatin (300 mg/m2) during CTRT has been demonstrated to impact on patients' outcome. The aim of the current study is to investigate whether the IC could impact on cisplatin dose intensity during CTRT in a homogeneous population of OPC. Materials and methods The study population consisted of 101 pts with stage III-IV OPC treated from 07/2004 to 12/2011 with IC (n = 53) plus CTRT or CTRT alone (n= 48). IC consisted of TPF, while weekly 50 mg/m2 or 3-weekly 100 mg/m2 cisplatin was administered concurrently with RT (planned chemotherapy dose = 300 mg/m2; planned RT dose = 66-70 Gy, 3DRT or IMRT with a conventional or accelerated fractionation). Carboplatin substituted cisplatin in case of creatinine clearance less than 60 mL/min. HPV status, concurrent cisplatin dose intensity and RT overall treatment time (OTT) were analyzed. Results Stage IV pts were 100% in the IC group and 83% in CTRT, while HPV positive OPCs were detected in 58% and 63% of the cases respectively. TPF median number of cycles was 3, with induction cisplatin median dose administered of 200 mg/ m2. RT median total dose administered was 70 Gy in both group. Accelerated fractionated RT was adopted in 65% of CTRT alone group and in 21% of IC group. Two pts (1 in each group) did not complete CTRT because of cardiovascular events. Mean and median dose intensity of cisplatin concurrent to RT in the IC group was 77.7% and 75% (35%-100%) while in CTRT group was 86% and 91.7% (33%-100%), with a p value = 0.014. In 11 pts (21%) treated with IC and only in 1 pts (2%) treated with CTRT alone cisplatin was substituted with carboplatin. No different cisplatin dose intensity was identified in HPV positive versus HPV negative cases. Prolonged OTT (longer than 5 days) was observed in two patients in IC group due to sepsis and severe mucositis as well as in one patient in CTRT group due to machine failure. Conclusions Concurrent cisplatin dose intensity was significantly reduced in pts receiving IC compared to CTRT alone, irrespective of HPV status. Whether this has an impact on the results of IC followed by full dose CT/RT has to be clarified. Disclosure All authors have declared no conflicts of interest.

Frequency and genotype distribution of multiple human papillomavirus infections in cancer of the head and neck in a Mexican population

The aim of this study was to determine the frequency and genotype distribution of single and multiple human papillomavirus (HPV) infections in head and neck squamous cell carcinomas (HNSCCs) in a Mexican population and to assess their associations with smoking and drinking habits and clinicopathologic characteristics. A cross-sectional study was performed on a sample of patients diagnosed with HNSCCs. Tumor DNA was amplified using polymerase chain reaction with HPV consensus and multiplex primers. The associations among HPV status, survival, and clinical characteristics were analyzed. Sixteen of the 43 HNSCCs were HPV positive. HPV16 was the most prevalent type, with single infections present in 5 cases, whereas another 5 cases were combined with HPV56 infection. There was a significant association between HPV infection and oropharyngeal cancers. HPV positivity was associated with overall survival at a nearly significant P level of 0.06. Our data support the importance of HPV infection in oropharyngeal cancer, with a trend toward higher survival in HPV-positive cases.

Equivocal p16 Immunostaining in Squamous Cell Carcinoma of the Head and Neck: Staining Patterns are Suggestive of HPV Status

p16 immunohistochemistry (IHC) is commonly used as a surrogate marker for human papillomavirus (HPV) detection in squamous cell carcinomas of the head and neck (SCCHN). However, the HPV status of tumors not staining strongly for p16 is difficult to interpret and may require the use of PCR, not available in all laboratories, as a final arbiter. We aim to determine if staining pattern in equivocal p16 staining and correlation to the percentage of positively stained tumor cells is predictive of HPV status. A retrospective review was performed on all SCCHN that underwent p16 IHC and PCR in our institution from 2007 to 2010. Descriptors of staining pattern in the original IHC report were retrieved. All available IHC slides were reviewed and reclassified using consensus staining pattern descriptors. Original and reclassified descriptors were compared to the final PCR HPV status for statistical significance using the χ(2) test. An estimate of the percentage of tumor cells that showed any form of staining was performed. Thirty-two SCCHN cases that underwent PCR HPV testing had equivocal p16 IHC results. Twenty-six cases available for review were reclassified into four staining patterns. Comparing age, sex, tumor site and diagnosis to HPV PCR status showed no statistically significant findings. However, comparing original descriptors to HPV status was statistically significant with isolated staining associated with negative HPV status (p=0.0002). Analysis using reclassified descriptors showed strong association of membranous/cytoplasmic staining of isolated cells with negative HPV status and faint, diffuse nuclear and cytoplasmic staining with positive HPV status (p=0.00006). HPV-negative cases with the former pattern had no more than 30% positively-stained tumor cells and HPV-positive cases with the latter pattern had 50-90% positively-stained cells. Our results suggest that pattern of staining in p16 IHC is associated with HPV status. For instance, a diffuse nuclear and cytoplasmic staining pattern, regardless of intensity, is associated with HPV positivity. The HPV-positive cases determined by staining pattern were also associated with a higher percentage of stained tumor cells.

Detection of human papillomavirus DNA in fine needle aspirates of women with breast cancer

Background: HPV infection is the most commonly distributed sexually transmitted disease. Human papillomavirus has also been linked to malignant tumors of many human organs. The presence of viral DNA in breast cancer cells is controversial. The aim of the present study was to investigate the presence of HPV-DNA in a group of Greek women with breast carcinoma. Methods: Liquid cytology specimens from 35 malignant breast cases and 35 cases with benign breast lesions were investigated by PCR (clinical arrays technique). In addition, in situ hybridization was performed on all HPV positive cases. Results: HPV-DNA was detected in 17.14% of the carcinoma cases and HPV16 DNA was present in 83.3% of them. All benign breast lesions were negative for HPV-DNA. Conclusion: Our report confirmed the presence of HPV in breast cancer cells while the most prevalent type was HPV16. More studies are necessary in order to elucidate the pathogenesis of HPV and a possible way of prevention of some breast cancers.

Correlation between ebv co-infection and HPV16 genome integrity in Tunisian cervical cancer patients

Infection with high risk Human papillomavirus (HR-HPV) is necessary but not sufficient to cause cervical carcinoma. This study explored whether multiple HR-HPV or coinfection with Epstein-Barr virus (EBV) influence the integration status of HPV16 genome. The presence and typing of HPV in a series of 125 cervical specimens were assessed by polymerase chain reaction (PCR) using the specific primers for the HPV L1 region. As for EBV infection, the viral EBNA1 gene was used for its detection through PCR amplification. Disruption of the HPV E2 gene was assessed by amplification of the entire E2 gene with single set of primers, while E2 transcripts were evaluated by a reverse transcription PCR method (RT-PCR). The overall prevalence of HPVDNA was of 81.8% in cervical cancers versus 26.9% in benign lesions. In HPV positive cases, HPV16 and HPV18 were the most prevalent types, followed by HPV types 33, 31. EBV EBNA1 prevalence was statistically more frequent in cervical carcinomas than in benign lesions (29.5%, vs 9.6%; P=0.01). No viral infection was detected in healthy control women. The uninterrupted E2 gene was correlated with the presence of E2 transcripts originating from the HPV episomal forms. It was observed that integration was more common in HPV18 and EBV coinfection. The presence of EBV caused a five-fold [OR= 5; CI= 1.15-21.8; P = 0.04] increase in the risk of HPV16 genome integration in the host genome. This study indicates that EBV infection is acting as a cofactor for induction of cervical cancer by favoring HPVDNA integration.

HPV Analysis in Distinguishing Second Primary Tumors From Lung Metastases in Patients With Head and Neck Squamous Cell Carcinoma

For patients with head and neck squamous cell carcinoma (HNSqCC), the development of squamous cell carcinoma (SqCC) in the lung may signal a new primary or the onset of metastatic dissemination. Although the distinction influences prognosis and therapy, it may not be straightforward on histologic or clinical grounds. The human papillomavirus (HPV) is an etiologic agent for SqCCs arising from the oropharynx but not for SqCCs arising from other head and neck sites. For patients with HNSqCC who develop a lung SqCC, HPV analysis could be useful in establishing tumor relationships. High-risk HPV in situ hybridization was performed on 54 lung SqCCs from patients with a previously diagnosed HNSqCC and on 166 primary lung carcinomas from patients without a prior HNSqCC. HPV was detected in 11 of 220 (5%) cases. All HPV-positive cases were from patients with a prior oropharyngeal SqCC. For the paired oropharyngeal and lung SqCCs, HPV status was concordant in 95% of cases. Time from treatment of HPV-positive oropharyngeal carcinomas to detection of lung carcinoma ranged from 1 to 97 months (mean, 36 mo). Two HPV-positive cancers were detected in the lung 8 years after treatment of the oropharyngeal primary. Despite the long interval, E6 sequencing analysis of 1 of these paired samples confirmed that the tumors harbored the same HPV-16 variant. HPV does not seem to play a role in the development of primary lung cancer. For patients with oropharyngeal SqCC who develop lung SqCC, HPV analysis may be helpful in clarifying tumor relationships. These relationships may not be obvious on clinical grounds, as HPV-related HNSqCC may metastasize long after treatment of the primary tumor.

The epidemiology of anal cancer

Anal cancer comprises malignancies of the anal canal principally of two morphologic variants: squamous cell carcinoma (SCC) and adenocarcinoma. In most settings, SCC compromises more than 70% of cases. In the general population, anal cancer is uncommon, with age-standardised incidence rates mostly between 1 and 2 per 100000 per year. However, incidence of anal SCC is increasing by 1-3% per year in developed country settings. High-risk human papillomavirus (HPV) types can be detected in 80-90% of all anal SCC cases, making it second only to cervical cancer in the closeness of its association with this virus. HPV-16 can be detected in ~90% of HPV-positive cases of anal SCC. Case-control studies have demonstrated that sexual risk factors (homosexuality in men and multiple sexual partners in women) are strongly associated with anal cancer risk. Other risk factors include immune deficiency and tobacco exposure. Anal cancer rates are highest in homosexual men, particularly in those who are HIV-positive, in whom anal cancer is among the most common of all cancers. Vaccination against HPV holds great promise for anal cancer prevention for those not already HPV-infected. For the current generation of adult high-risk populations, screening programs to allow early detection and treatment are under investigation.

Human papillomavirus genotype distribution in cervical cancer cases in Spain. Implications for prevention

ObjectiveHuman papillomavirus (HPV) genotype distribution in invasive cervical cancer (ICC) is critical to guide the introduction and to assess the impact of HPV prophylactic vaccines. This study aims to provide specific information for Spain. Methods1043 histological confirmed ICC cases diagnosed from 1940 to 2007 from six Spanish regions were assembled. HPV DNA detection was performed by SPF10 broad-spectrum PCR followed by deoxyribonucleic acid enzyme immunoassay and genotyping by reverse hybridization line probe assay (LiPA25) (version 1). ResultsOf 1043 ICC cases, 904 were HPV DNA positive (adjusted prevalence: 89.1%). The eight most common types, in decreasing order, were HPV 16, 18, 33, 31, 45, 35, 52 and 56, accounting for more than 90% of cases. HPV 16 and 18 contributed to 72.4% of all HPV positive ICC cases. In cervical adenocarcinomas, this contribution increased up to 94%. HPV 16 and 18 relative contributions showed a stable pattern over the 60year study period. HPV 45, 18 and 16-positive ICC cases presented at younger ages than cases with other HPV types (adjusted mean age: 43.8, 45.2, 52.6 and 57.7years, respectively). ConclusionsHPV 16 and 18 accounted together for a 72.4% of positive cases, with no statistically significant changes in their relative contributions over the last decades. In 94% of cervical adenocarcinomas we identified at least one of the two HPV types included in the current vaccines (HPV 16/18). Results suggest a major impact of HPV vaccines on reduction of ICC burden in Spain in the HPV vaccinated cohorts.

Human papillomavirus detected in esophageal squamous cell carcinoma in Iran

IntroductionConsidering the different results obtained regarding the association of HPV with the development of esophageal squamous cell carcinoma (ESCC) in different populations, we aimed to determine the frequency of HPV infection and its subtypes in ESCC in Iranian patients. MethodA total of 100 paraffin-embedded tissue samples of ESCC diagnosed during 1991 and 2005 in the Institute of Cancer affiliated to Tehran University of Medical Sciences were selected. Seven out of 100 samples were excluded due to low quality of extracted DNA from paraffin-embedded specimens. Thus, 93 samples were included for HPV genotyping. ResultAll samples were examined using SPF10 primers for HPV detection. HPV DNA was positive in 8 out of 93 (8.6%) ESCC specimens. Using INNO-LiPA genotyping system we detected the genotypes of 5 out of 8 HPV-positive samples. Both HPV types 16 and 6 were detected in 3 specimens; one sample was positive for HPV type 18 and 2 samples were co-infected with two HPV types. There were no statistically significant differences between HPV-positive and HPV-negative cases with regard to clinical and pathologic findings. Three samples were positive for SPF10 indicating HPV infection; however, the exact HPV type could not be clarified using INNO-LiPA genotyping . ConclusionIn conclusion, the present study showed that a small proportion of ESCC specimens from Iran harbor HPV16, 18 genome using a highly sensitive method. As different rates have been reported from Iran, a more widespread study with more precise definition of geographical differences could delineate the potential involvement of HPV in the development of ESCC in Iranian population.

Low Prevalence of Human Papillomavirus (HPV) in Chinese Patients with Breast Cancer

This retrospective study investigated the presence of human papillomavirus (HPV) in Chinese women with breast cancer, and the correlation between HPV infection and carcinogenesis. Tumour and non-cancerous breast tissue samples were obtained from 62 female patients with breast cancer; normal breast tissue samples were obtained from 46 women without breast cancer. HPV DNA was detected by nested polymerase chain reaction using consensus primers; HPV subtypes were determined by reverse dot blot and pyrosequencing analyses. HPV was found in tumour tissue samples from four of the 62 patients (6.5%), while no HPV DNA was detected in either the non-cancerous samples from patients with breast cancer or from the normal breast tissue controls. Of the four HPV-positive cases, three were HPV 16 positive (75%) and one was HPV 18 positive (25%). The low frequency of HPV detected in this study suggests that this infection is not a major risk factor in breast cancer development.

Evaluation of Human Papilloma Virus Diagnostic Testing in Oropharyngeal Squamous Cell Carcinoma: Sensitivity, Specificity, and Prognostic Discrimination

Human papillomavirus-16 (HPV16) is the causative agent in a biologically distinct subset of oropharyngeal squamous cell carcinoma (OPSCC) with highly favorable prognosis. In clinical trials, HPV16 status is an essential inclusion or stratification parameter, highlighting the importance of accurate testing. Fixed and fresh-frozen tissue from 108 OPSCC cases were subject to eight possible assay/assay combinations: p16 immunohistochemistry (p16 IHC); in situ hybridization for high-risk HPV (HR HPV ISH); quantitative PCR (qPCR) for both viral E6 RNA (RNA qPCR) and DNA (DNA qPCR); and combinations of the above. HPV16-positive OPSCC presented in younger patients (mean 7.5 years younger, P = 0.003) who smoked less than HPV-negative patients (P = 0.007). The proportion of HPV16-positive cases increased from 15% to 57% (P = 0.001) between 1988 and 2009. A combination of p16 IHC/DNA qPCR showed acceptable sensitivity (97%) and specificity (94%) compared with the RNA qPCR "gold standard", as well as being the best discriminator of favorable outcome (overall survival P = 0.002). p16 IHC/HR HPV ISH also had acceptable specificity (90%) but the substantial reduction in its sensitivity (88%) impacted upon its prognostic value (P = 0.02). p16 IHC, HR HPV ISH, or DNA qPCR was not sufficiently specific to recommend in clinical trials when used in isolation. Caution must be exercised in applying HPV16 diagnostic tests because of significant disparities in accuracy and prognostic value in previously published techniques.

HPV E6/E7 mRNA Expression in Oropharyngeal Carcinoma

Objective: Although patients with human papillomavirus (HPV)-positive oropharyngeal carcinoma (OC) respond well to radiation therapy and have good prognosis, HPV integration in OC has not yet been fully analyzed. Therefore, we aimed to clarify the correlation between HPV infection or integration and prognosis of OC in a prospective study setting. Method: From 2006 to 2010, 56 patients with OC participated in the study. All patients were treated with 70 Gy concurrent chemoradiotherapy (CCRT). If OC did not show a partial or complete response at 40-Gy irradiation, radical resection for residual tumor was performed. Results: A total of 26 patients (46.4%) had HPV-positive OC, and 70% of HPV-16-positive OC cases showed integration with the host genome. Control samples obtained from chronic tonsillitis patients showed 1 of 47 specimens as positive for HPV. Patients with high HPV load also showed E6/E7 mRNA expression. Recurrent-free survival at 3 years was 100% in HPV-positive cases and 88.9% in HPV-negative cases. Cases with HPV integration or E6/E7 mRNA expression tended to respond well to CCRT in spite of advanced tumor extensions. Conclusion: Although HPV testing is useful for OC, HPV integration or E6/E7 mRNA expression is a more specific marker for predicting a fair response to CCRT and good prognosis. These findings also indicate that HPV plays a key role in the development and extension of OC.

Newly developed liquid-based cytology. TACAS™: cytological appearance and HPV testing using liquid-based sample

Cell profiles determined by the thin-layer advanced cytology assay system (TACAS™), a liquid-based cytology technique newly developed in Japan, were analyzed in this study. Hybrid capture 2 (HC-2) was also performed using the liquid-based samples prepared by TACAS to ascertain its ability to detect human papillomavirus (HPV). Cell collection samples from uterine cervix were obtained from 359 patients and examined cytologically. A HC-2 assay for HPV was carried out in the cell specimens. All specimens were found to show background factors such as leukocytes. After excluding the 5 unsatisfactory cases from the total 354 cases, 82 cases (23.2%) were positive and 272 cases (76.8%) were negative for HPV. Cell specimens from 30 HPV-positive cases and 166 HPV-negative cases were subjected to 4weeks of preservation at room temperature. Then, when subsequently re-assayed, 28 cases (93.3%) in the former group were found to be HPV positive and 164 cases (98.8%) in the latter group were found to be HPV negative. These results supported the excellent reproducibility of TACAS for HPV testing. A reasonable inference from the foregoing analysis is that TACAS may be distinguished from other liquid-based cytological approaches, such as ThinPrep and SurePath, in that it can retain the cell backgrounds. Furthermore, this study raises the possibility that cell specimens prepared using TACAS could be preserved for at least 4weeks prior to carrying out a HC-2 assay for HPV.

Single and multiple human papillomavirus infections in cervical abnormalities in Portuguese women

Persistent infection with high-risk (HR) human papillomavirus (HPV) types is necessary for cervical cancer development. However, little is known about the influence of multiple HPV infections on cervical lesion risk. The aim of this study was to evaluate the frequency of single and multiple HPV infections in Portuguese women, and to assess the frequency of multiple infections in cervical intraepithelial neoplasia (CIN). HPV prevalence, type-specific prevalence and extent of multiple infections were assessed in 1057 cervical samples. The Clinical Array HPV assay was used to detect 35 HPV types. According to histological diagnosis, 425 samples were normal, 375 were CIN1, and 257 were CIN2+. HPV status was studied in relation to age and lesion severity. The prevalence of HPV infection was 52.7%; 25.4%, 67.2% and 76.7% were positive for any HPV type in the normal, CIN1 and CIN2+ cases, respectively. Among HPV-positive cases, 32.0% were associated with multiple infections. Among multiple infections, 96.1% harboured HR HPV types and 38.2% HR–low risk (LR) HPV types. Overall, 33 different HPV types (18 HR and 15 LR) were detected. HR HPV types (44.1%) were significantly more prevalent than LR HPV types (8.6%). The most frequent genotype was HPV 16 (25.5%), followed by HPV 31, 53, 66, 58, and 51. Multiple infections showed a significant increase (p 0.005) according to severity of neoplasia, particularly for HR–HR HPV infections (p 0.003). No association between age and multiple HPV infections was observed (p 0.812). However, multiple HR HPV infections were more frequent in women under 30 years of age (35.3%).

Predictive value of human papillomavirus (HPV) 16 in situ hybridization (ISH) and p16 immunohistochemistry (IHC) for HPV oncogene expression in paraffin-embedded oropharynx squamous cell carcinoma (OPSCC).

5512 Background: Tumor HPV status is a strong, independent predictor of OPSCC survival and increasingly used for therapeutic decision making. Methods for classification are variable and not standardized. Here we validate our methods for HPV classification used in past and ongoing cooperative group trials (ECOG and RTOG). Methods: Consecutive OPSCC diagnosed from 2000-2009 were obtained from four referral centers. Tumor DNA and RNA were extracted and categorized as evaluable or inevaluable by DNA detection (ERV3) and mRNA expression (RPLPO) of control genes by quantitative (q) PCR. Presence of 18 high-risk HPV types was detected by consensus primer PCR and line-blot hybridization (Inno-Lipa) followed by viral load measurement and HPV oncogene expression (E6/7) analysis by HPV type-specific q RT-PCR. P16 expression was detected by IHC and HPV16 by ISH and sensitivity (S), specificity (SP), positive (PPV) and negative predictive value (NPV) were compared to gold standard HPVE6/7 oncogene expression. Assay interpretation was performed by three masked pathologists. Inter-assay and inter-rater agreement was evaluated by kappa statistic. Sample size provided 80% power for Cohen’s kappa of 0.8 or higher among three raters. Results: 235/237 OPSCC were evaluable, 183 (78%) HPV DNA-positive, and 158 (67%) HPVE6/7 expression positive (16, 92%; 18, 3%; 33, 3%; 35, 1%; 58, 1%) with median viral load of 15.7 copies/cell, E6/7 expression of 210 copies/1,000 RPLPO, p16 expression 46.7 vs. 2.9 copies/1000 RPLPO for HPVE6/7-negative, p<0.001. S/SP/PPV/NPV of HPV16 ISH for HPV16E6/7 was 91/91/94/86 and for p16 for HPVE6/7 was 97/82/92/93 and for combined testing was 84/97/99/75. P16 IHC and HPV16 ISH had very good agreement (kappa=0.64) with HPVE6/7 expression detected in 99%/63%/50%/3% of ++/+-/-+/-- cases, respectively. Inter-rater agreement was excellent for p16 (kappa=0.95-.98) and HPV16 ISH (kappa.0.83-.91). Conclusions: HPV16 ISH and p16 IHC have excellent assay performance. However, if results are used for therapy “deintensification” among HPV-positive cases, combined testing provides higher SP and PPV.

Human papillomavirus (HPV) and rising oropharyngeal cancer incidence and survival in the United States.

5529 Background: Recent increases in incidence and survival of oropharyngeal cancers in the United States (U.S.) have been attributed to human papillomavirus (HPV) infection, but empirical evidence is lacking. Methods: HPV status was determined for 271 oropharyngeal cancer cases (1984-2004) collected by population-based cancer registries of the National Cancer Institute’s Surveillance, Epidemiology, and End Results program in Hawaii, Iowa, and Los Angeles. Tumor HPV status was determined by PCR and genotyping (Inno-LiPA), HPV16 viral load, HPV16 mRNA expression, and HPV16 in situ hybridization. Trends in HPV prevalence across four calendar periods were estimated using logistic regression. Survival of HPV-positive and HPV-negative cases was compared by Kaplan-Meier and multivariate Cox regression analyses. Results: HPV prevalence in oropharyngeal cancers significantly increased over calendar time regardless of HPV detection assay (p-trend<0.05). For example, HPV prevalence by Inno-LiPA increased more than ...

Human papillomavirus infection in nasal polyps in a Chinese population

In order to determine the prevalence and genotype distribution of human papillomavirus (HPV) infection in patients with nasal polyps, a total of 204 patients with nasal polyps and 36 healthy controls were recruited for this study. Genomic DNA was extracted from paraffin-embedded tissue sections. HPV DNA genotyping was achieved by a flow-through hybridization and gene-chip method. HPV-positive infection was identified in 82 of 204 (40.2 %) patients, while HPV DNA was not found in healthy controls (P<0.05). Genotyping analysis showed that low-risk HPV genotype 11 was the most prevalent type of HPV in nasal polyps (45.28 %). Both single and multiple HPV genotype infections were found in these HPV-positive cases, although most (74.39 %) were infected with a single genotype. In addition, there was no correlation between HPV infection or HPV subtypes and the clinicopathological characteristics of patients, such as age, gender, number of surgery and disease course. The data from our study clearly demonstrated that HPV infection was associated with nasal polyps. Both high-risk HPV and low-risk HPV (LR-HPV) genotypes were identified in nasal polyp tissues, and LR-HPV-11 was the most prevalent type. Future research will explore the association of HPV infection with the development and progression of nasal polyps.