Positive Human Papillomavirus Status Research Articles

Adequacy of fine-needle aspiration specimens for human papillomavirus infection molecular testing in head and neck squamous cell carcinoma

Background:Head and neck squamous cell carcinoma is often associated with human papillomavirus (HPV) infection. Positive HPV status has been associated with increased response to treatment and improved prognosis in terms of recurrence-free and overall survival. In certain instances, diagnosis is performed through fine-needle aspiration of lymph nodes with metastatic carcinoma, often demonstrating extensive tumor necrosis. We evaluated the effect of tumor necrosis on deoxyribonucleic acid (DNA) adequacy for HPV molecular testing.Materials and Methods:Retrospective review of the pathology files at our institution identified cases of squamous cell carcinoma (SCC) diagnosed by fine-needle aspiration (FNA) on which HPV DNA molecular testing was performed. The cases were classified according to percent tumor necrosis into three categories (<10% necrosis, 10-70% necrosis and >70% necrosis) and the percentage of cases with adequate HPV DNA for molecular testing in each of the categories was compared. When available, p16 immunohistochemistry performed on the cases was compared with HPV status by molecular testing.Results:A total of 70 cases from 67 patients were included in the study. Adequate DNA for molecular HPV testing was obtained from samples of 47 cases (67%) while samples from 23 cases (33%) were inadequate for molecular testing. Of the adequate samples, 36 (77%) were positive and 11 (23%) were negative for high-risk HPV. Adequate DNA for testing was obtained in 22 out of 33 cases showing no necrosis (67%), 10 out of 16 cases showing partial necrosis (63%) and in 13 out of 17 cases showing extensive necrosis (76%).Conclusion:Our study found that HPV molecular testing is not influenced by percent tumor necrosis or method by which FNA was performed. We believe that a portion of the FNA specimen obtained from head and neck lesions diagnosed as SCC during the rapid on-site evaluation should be sent for HPV DNA testing, independent of the amount of tumor necrosis, thus guaranteeing availability of specimen for HPV testing.

Prognostic value of human papillomavirus and squamous cell carcinoma antigen in head and neck squamous cell carcinoma

To clarify the synergistic influence of human papillomavirus (HPV) status and squamous cell carcinoma antigen (SCCA) mRNA expression on head and neck squamous cell carcinoma (HNSCC) prognosis, HPV DNA presence and SCCA1 and SCCA2 mRNA expression were determined by PCR and quantitative real-time RT-PCR, respectively, in 121 patients with primary HNSCC who were receiving curative treatment. HPV DNA was detected in 28.1% (34/121) of HNSCC cases, and only high-risk types (HPV-16, HPV-33, HPV-35 and HPV-58) were observed. Positive HPV status showed a significantly better prognosis than negative HPV status (P = 0.022). An elevated SCCA2/SCCA1 mRNA ratio was an independent predictor of disease recurrence (P = 0.004). In addition, HPV-negative patients with a high SCCA2/SCCA1 ratio (>0.27) had a significantly lower recurrence-free survival rate than HPV-negative patients with a low SCCA2/SCCA1 ratio (P < 0.011). Our findings revealed that both HPV status and the SCCA2/SCCA1 mRNA ratio are independently associated with prognosis in HNSCC. Patients with both a HPV-negative status and a high SCCA2/SCCA1 ratio might need intensified treatment and rigorous follow up after treatment because of the high risk of recurrence.

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.

Importance of HPV involvement and FOXP3+ T-cell status as prognostic factors in tonsilar squamous cell carcinoma.

5586 Background: Human papillomavirus (HPV) status is a strong and independent favorable prognostic factor for survival in tonsilar squamous cell cancer (TSCC). The reason for the improved survival in HPV associated TSCC is unclear. Recently, activation of immune surveillance mechanism against non-self Ag is postulated to one of reasonable causes as favorable prognosis of HPV associated with TSCC. Methods: We reviewed the medical records of histologically confirmed locally advanced TSCC patients, curatively treated in Asan Medical Center from January 2000 to December 2008. The immunohistochemistry (IHC) assays for p16 and FOXP3 were done in TSCC pafaffin-embedded samples. Results: We identified 79 patients who met the inclusion criteria. The median age was 54 years (range 32-76) and 16 (20%) patients were stage III and the others were all stage IV. With the median follow up of 62.9 months (95% CI, 59.2 - 66.7), sixty three (80%) were HPV-positive with p16 overexpression, and 38 (48%) were Treg-positive with FOXP3. Treg involvement was significantly related to HPV positive status (P=0.011). The result was the same after adjustment of age, T &amp; N stage, smoking exposure and alcohol consumption. (Odd ratio = 6.54, 95% CI 1.58-27.1, P=0.01) Five-year overall survival (OS) rate in HPV-positive group was significantly higher than that of HPV-negative group (78% and 63%, Hazard Ratio (HR)=0.347, 95% CI 0.14-0.87, P=0.025), and 5-year OS of Treg-positive group was also higher than that of Treg-negative group (89% and 61%, HR=0.158, 95% CI 0.05-0.53, P=0.003). In multivariate analysis, the Treg status was an independent prognostic factor (HR=0.11, 95% CI 0.03-0.40, P=0.001), as well as HPV status. (HR=0.28, 95% CI 0.10 - 0.78, P=0.016). Conclusions: HPV positivity was associated with Treg positivity in TSCC and both were found to be favourable prognostic factors for survival.

ヒト乳頭腫ウイルス関連中咽頭扁平上皮癌におけるThymidylate synthase発現とその臨床像に関する検討

The focus of human papilloma virus (HPV), particulary HPV 16 is on the role of carcinogenic and prognostic factors on oropharyngeal squamous carcinoma (OSCC). However, it remains unclear why patients with HPV-positive tumors have better outcomes than those with HPV-negative tumors. Thymidylate synthase (TS) is one of the initial key enzymes in the 5-fluouracil (5-FU) metabolic pathway. Clinical studies showed that intratumoural TS level was related to the response to 5-FU-based chemotherapy in patients with several types of cancer such as gastroenterological and head and neck cancers. We investigated the prevalence of HPV infection and TS expression in the patients with OSCC and evaluated the prognostic implications according to the HPV status and TS expression. We evaluated for high-risk HPV types (HPV 16, 18, 31, 33, 51, 52, 58) using a real-time polymerase chain reaction (RT-PCR) assay on archival biopsies from 54 patients with OSCC. Immunohistochemical assessments for TS were also performed. HPV was positive in 22 (40.7%) of 54 samples. Of these positive cases, 21 (95%) carried HPV 16 and only 1 (5%) HPV58 sequences. TS was overexpressed in 25 (46.3%) of 54 samples. Of these, 19 (76.0%) had an HPV-negative status and 21 (84.0%) were heavy smokers. TS over-expression was associated with the patients with HPV-negative tumors (P = 0.02) and heavy smokers (p = 0.012). Univariate analysis revealed that HPV positive status (77.3% vs. 29.0%; p = 0.006) significantly improved overall survival. Conversely, no remarkable prognostic difference was observed on immunohistochemical analysis of TS expression. A multivariate analysis using Cox's proportional hazard model showed that early T stage (T1-2), early N stage (N0-1), and positive HPV status were significantly independent predictors for superior overall survival. Our studies suggested that positive HPV status was most strongly associated with a favorable prognosis in the patients with OSCC. TS expression has an unusual aspect as a biomarker for OSCC, though it was not related to prognosis.

Survival of patients with HPV-positive oropharyngeal cancer after radiochemotherapy is significantly enhanced

The purpose of this study was to evaluate the incidence and clinical significance of HPV (Human papilloma virus) infection in patients with head and neck cancer who had received radiotherapy in Eastern Austria. 88 patients with head and neck cancer including 26 patients with oral cavity cancer, 45 patients with oropharyngeal cancer, seven patients with laryngeal carcinoma and ten patients with carcinoma of the hypopharynx were screened for high risk HPV by immunohistochemistry, PCR (Polymerase Chain Reaction) and in-situ hybridization. 29 out of 45 patients with a squamous cell carcinoma of the oropharynx received radiotherapy alone, radiotherapy in combination with cisplatin or cetuximab. Of the investigated 29 patients with oropharyngeal cancer receiving conservative treatment, 11 had a HPV-positive and 18 a HPV-negative tumor. Patients received radiation ± cisplatin or cetuximab, where the HPV-positive patients had a significant better response to treatment and overall survival (p = 0.015) as well as disease-free survival (p = 0.001) after therapy. Patients with oropharyngeal carcinoma and a positive HPV status respond considerably better to radiochemotherapy than patients with HPV-negative tumors. HPV screening is a simple procedure and can easily be implemented in routine pathology investigations and should be included in standard operational procedures for the diagnosis and therapy of head and neck cancer patients.

Attitudes Toward HPV Testing: Interview Findings From a Random Sample of Women in Ontario, Canada

As most women diagnosed with cervical carcinoma have been “inadequately screened,” improvements in screening are critical. After abnormal Pap test findings (through liquid-based cytology), residual specimens now can be tested simultaneously for oncogenic types of Human Papilloma virus (HPV). If these “reflex” HPV tests are negative, Pap tests need not be repeated for 12 months. Women with positive oncogenic HPV tests, however, can be referred immediately for colposcopy. There has been concern that “stigma” issues could be associated with positive HPV status (because of its sexual transmission) that might cause women to avoid this reflex HPV testing. We addressed this concern by assessing whether stigma issues surface in relation to HPV testing. We randomly selected 20 women and administered to them semistructured telephone interviews that included responses to a scenario of reflex HPV–DNA testing. Interview transcripts were analyzed qualitatively. Highly limited knowledge levels were found about HPV, but, following education about screening options, there was no rejection of HPV testing. In conclusion, it appears that women favor reflex HPV testing due to its “convenience” and perceptions that it is “the least intrusive option more definitive than Pap testing.”

Negative Predictive Value of Human Papillomavirus Test Following Conization of the Cervix Uteri

Objective. The goal of this study was to determine/evaluate the negative predictive value of human papillomavirus (HPV) testing following conization of cervix uteri.Methods. A prospective analysis was undertaken on 79 cone biopsies of women with high-grade lesions (cervical intraepithelial neoplasia (CIN) III). HPV testing was performed on cervical smears before and after conization. We correlated the margin status (defined as positive cone margin or endocervical curettage status) and positive conization HPV status with the residual disease in a hysterectomy specimen. A Digene II kit was used to perform HPV testing. HPV detection was done by Hybrid Capture assay.Results. Of the 79 patients, 47(59.5%) had positive margins after conization. HPV testing was positive in 37 cases (78.7%) and negative in 10 cases (21.3%). Residual disease was found in 31 of 47 (66%) postconization hysterectomy specimens. No residual lesions were found in HPV-negative cases. Of the 32 cases with negative margins following conization, HPV testing was negative in 25 cases (78%) and was positive in 7 cases (22%). Among these 25 cases with negative HPV tests, no residual lesion was detected, and in 7 HPV-positive cases, only one residual lesion was found.Conclusion. HPV testing is potentially an effective tool in predicting residual dysplasia after conization and could potentially assist in the decision between hysterectomy and conservative follow-up in women with CIN III.