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Objective: To investigate the prevalence, genotypes, and prognostic values of Epstein-Barr virus (EBV) and human papillomavirus (HPV) infections in Japanese patients with different types of head and neck cancer (HNC). Methods and Materials: HPV and EBV DNA, EBV genotypes and LMP-1 variants, and HPV mRNA expression were detected by PCR from fresh-frozen HNC samples. HPV genotypes were determined by direct sequencing, and EBV encoded RNA (EBER) was examined by in situ hybridization. Results: Of the 209 HNC patients, 63 (30.1%) had HPV infection, and HPV-16 was the most common subtype (86.9%). HPV E6/E7 mRNA expression was found in 23 of 60 (38.3%) HPV DNA-positive cases detected. The site of highest prevalence of HPV was the oropharynx (45.9%). Among 146 (69.9%) HNCs in which EBV DNA was identified, 107 (73.3%) and 27 (18.5%) contained types A and B, respectively, and 124 (84.9%) showed the existence of del-LMP-1. However, only 13 (6.2%) HNCs were positive for EBER, 12 (92.3%) of which derived from the nasopharynx. Co-infection of HPV and EBER was found in only 1.0% of HNCs and 10.0% of NPCs. Kaplan-Meier survival analysis showed significantly better disease-specific and overall survival in the HPV DNA+/mRNA+ oropharyngeal squamous cell carcinoma (OPC) patients than in the other OPC patients (P = 0.027 and 0.017, respectively). Multivariate analysis showed that stage T1–3 (P = 0.002) and HPV mRNA-positive status (P = 0.061) independently predicted better disease-specific survival. No significant difference in disease-specific survival was found between the EBER-positive and -negative NPC patients (P = 0.155). Conclusions: Our findings indicate that co-infection with HPV and EBV is rare in HNC. Oropharyngeal SCC with active HPV infection was related to a highly favorable outcome, while EBV status was not prognostic in the NPC cohort. [ABSTRACT FROM AUTHOR]

Background: The clinical importance of serum squamous cell carcinoma antigen (SCCA) and SCCA subclasses has not been established for treating inverted papilloma (IP). The aim of this study was to clarify the clinical importance of serum SCCA and its subclasses in IP, compared with maxillary squamous cell carcinoma and inflammatory disease. Methods: Serum SCCA was measured in 22 patients with IP (IP group), 11 with maxillary squamous cell carcinoma (carcinoma group), and 22 with inflammatory disease (inflammatory group). mRNA expression of SCCA subclasses was examined using quantitative real-time polymerase chain reaction. Results: In the IP group, 81.8% showed elevated serum SCCA, and 90.3% with recurrent IP showed elevated SCCA. The preoperative SCCA value (mean ± SD, 3.99 ± 4.39) in the IP group was significantly higher than in the carcinoma (1.28 ± 0.88; p = 0.012) and inflammatory (0.60 ± 0.31; p < 0.001) groups. mRNA expression of SCCA1 and SCCA2 in the IP group was higher than in the carcinoma and inflammatory groups. The SCCA2/SCCA1 ratio of mRNA expression (0.11 ± 0.06) in the IP group was similar to that (0.11 ± 0.09) in the inflammatory group, although the ratio (0.20 ± 0.12) in the carcinoma group was significantly higher than in the IP and inflammatory groups. The receiver operating characteristic curve analysis for the SCCA2/SCCA1 ratio to detect carcinoma yielded an area under the curve of 0.760 (95% confidence interval, 0.626-0.894). Conclusion: The serum level of SCCA is effective for detecting IP, including recurrent IP. In contrast, the SCCA2/SCCA1 ratio is useful for detecting squamous cell carcinoma among other sinonasal diseases. [ABSTRACT FROM AUTHOR]

Laryngeal papilloma (LP), which is associated with infection by human papillomavirus (HPV)-6 or -11, displays aggressive growth. The precise molecular mechanism underlying the tumorigenesis of LP has yet to be uncovered. Building on our earlier research into HPV-6, in this study, the viral gene expression of HPV-11 was investigated by quantitative PCR and DNA/RNA in situ hybridization. Additionally, newly developed antibodies against the E4 protein of HPV-6 and HPV-11 were evaluated by immunohistochemistry. The average viral load of HPV-11 in LP was 1.95 ± 0.66 × 105 copies/ng DNA, and 88% of HPV mRNA expression was found to be E4, E5a, and E5b mRNAs. According to RNA in situ hybridization, E4 and E5b mRNAs were expressed from the middle to upper part of the epithelium. E4 immunohistochemistry revealed a wide positive reaction in the upper cell layer in line with E4 mRNA expression. Other head and neck lesions with HPV-11 infection also showed a positive reaction in E4 immunohistochemistry. The distribution pattern of HPV DNA, viral mRNA, and E4 protein in LP with HPV-11 infection was quite similar to that of HPV-6. Therefore, it might be possible to apply these E4-specific antibodies in other functional studies as well as clinical applications, including targeted molecular therapies in patients with HPV-6 and HPV-11 infection. [ABSTRACT FROM AUTHOR]

The aim of our study was to investigate a possible correlation between Epstein-Barr virus (EBV) infection status, including its latent gene expression, and expression of allergy-related genes in human tonsil-derived cells. In the tonsil-derived cells from the patients undergoing routine tonsillectomies for palatine tonsil hypertrophy or tonsillar focal infection, the presence of EBV DNA and mRNA expressions of latent membrane protein (LMP)-1, Cε chain, and activation-induced cytidine deaminase (AID) were detected by PCR and RT-PCR analysis, respectively. Of all the 12 patients, PCR products amplified from EBV DNA BamHI W fragment were detected in the tonsils from the 10 patients (83.3%). LMP1 mRNA expressions were confirmed in the six patients (50%). Both LMP1 mRNA expressions and EBV DNA were detected in the five patients. EBV DNA, but not LMP1 mRNA expression, was detected in the five patients. LMP1 mRNA expression, but not EBV DNA, was detected in one patient. In one patient, neither EBV DNA nor LMP1 mRNA expression was confirmed. Cε mRNA expressions were confirmed in all the 12 patients along with AID mRNA expressions. The degree of Cε mRNA expression, however, varied with the patients. The Fisher’s exact probability test revealed a statistically significant correlation between LMP1 and Cε gene expressions, indicating that Cε mRNA expression level was significantly higher in the LMP1 positive samples than in the negative samples. On the other hand, there was no significant correlation between AID and LMP1 mRNA expressions. Thus, EBV infection is a notable factor capable of exacerbating allergic inflammation. [ABSTRACT FROM AUTHOR]

To evaluate the effect of expression of latent membrane protein (LMP) 1 encoded by Epstein-Barr virus (EBV) on Cε mRNA expression, mRNA levels were examined by RT-PCR or Northern blot analysis upon transient transfection of LMP1 in the splenocytes derived from Brown-Norway rats with or without immunization with 2,4-dinitrophenyl-conjugated Ascaris suum antigen. Splenocytes were transfected with LMP1 expression vector, pSG5-LMP1, using lipofection method. Cε mRNA levels were considerably increased by transfection with pSGS-LMP1 in the splenocytes derived from the nonimmunized rats; however, Cε mRNA levels were decreased in the splenocytes derived from the immunized rats. Cε mRNA expression in IgE-producing cells are modulated by LMP1, which might depend on the differentiation status of B cells upon exposure to allergen. [ABSTRACT FROM AUTHOR]

Laryngeal papilloma (LP) associated with human papillomavirus (HPV)-6 or -11 infection shows aggressive growth. However, the detailed molecular mechanism of virus-driven tumorigenesis has not been uncovered fully. HPV-6 viral gene expression and dynamic alterations were investigated with in situ localization of viral DNA and RNA in 13 patients with HPV-6-infected laryngeal papilloma. The average viral load was 4.80 × 105 ± 1.86 × 105 copies/ng DNA. E4, E5a, and E5b mRNAs accounted for 96% of the expression of 9 mRNAs. The alteration of viral DNA load during recurrence paralleled the mRNA expression levels, and the expression of all mRNAs showed a similar curve. E4, E5a, and E5b were expressed in the middle to upper part of the epithelium and were co-expressed in the same cells. E4 immunohistochemistry demonstrated an extensively positive reaction in the upper cell layer in accordance with E4 mRNA expression. These results suggest that individual viral genes are coordinately expressed for viral replication, virus release, and immunosurveillance avoidance. The newly developed E4-specific monoclonal antibody can be applied to further functional studies and clinical applications such as targeted molecular therapies. [ABSTRACT FROM AUTHOR]

Background: Oropharyngeal cancers associated with high-risk type human papillomavirus (HR-HPV) infection have better prognosis than virus negative cancers. Similarly, the HPV status in laryngeal cancer (LC) may be associated with better outcome. Methods: Samples from 88 patients with LC were investigated using the polymerase chain reaction (PCR) and p16 immunohistochemistry for HR-HPV analysis. The cut-off point for p16 overexpression was diffuse (≥75%) tumor expression with at least moderate (+ 2/3) staining intensity. Results: The 5-year cumulative survival (CS) rate was 80.7% in all patients with LC. According to a combination of HR-HPV DNA status and p16 overexpression, subjects with LC were divided into four groups: HR-HPV DNA-positive/p16 overexpression-positive (n = 5, 5.7%; CS = 100%), HR-HPV DNA-positive/p16 overexpression-negative (n = 11, 12.5%; CS =81.8%), HR-HPV DNA-negative/p16 overexpression-positive (n = 0), and HR-HPV DNA-negative/p16 overexpression-negative (n = 72, 81.8%; CS = 79.5%). HR-HPV DNA-positive/p16-positive cases tended to have integrated HPV infection and high viral load, compared with HR-HPV DNA-positive/p16 overexpression-negative cases. Conclusions: LC patients with HPV infection and high levels of p16 expression might have an improved survival outcome; however, it is necessary to recruit additional LC cases with HPV infection to determine the definitive characteristics of HPV-mediated LC and estimate survival outcome. These results may contribute to the development of a useful method for selecting patients with a potentially fair response to treatment and ensure laryngeal preservation. [ABSTRACT FROM AUTHOR]