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.

Department of Experimental Virology Institute of Hematology and Blood Transfusion 128 20 Prague Czech Republic

Department of Otorhinolaryngology and Head and Neck Surgery 1st Faculty of Medicine Charles University Prague University Hospital Motol 150 06 Prague Czech Republic

Department of Otorhinolaryngology and Head and Neck Surgery 1st Faculty of Medicine Charles University Prague University Hospital Motol 150 06 Prague Czech Republic ; Department of Experimental Virology Institute of Hematology and Blood Transfusion 128 20 Prague Czech Republic

Department of Otorhinolaryngology and Head and Neck Surgery 1st Faculty of Medicine Charles University Prague University Hospital Motol 150 06 Prague Czech Republic ; Institute of Microbiology Academy of Sciences of the Czech Republic Public Research Institution 142 20 Prague Czech Republic

Department of Pathology and Molecular Medicine 2nd Faculty of Medicine Charles University Prague 150 06 Prague Czech Republic

Department of Pediatric Hematology and Oncology 2nd Faculty of Medicine Charles University Prague University Hospital Motol 150 06 Prague Czech Republic

Institute of Microbiology Academy of Sciences of the Czech Republic Public Research Institution 142 20 Prague Czech Republic

Zobrazit více v PubMed

Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncology. 2009;45(4-5):309–316. PubMed

Zivile G, Janina D, Irina M, Saulius C, Raisa S, Aliaksandr Z. Prevalence of human papillomaviruses in patients with head and neck squamous cell carcinoma in Lithuania and Belarus. Journal of Medical Virology. 2014;86(3):531–535. PubMed

Hammarstedt L, Lindquist D, Dahlstrand H, et al. Human papillomavirus as a risk factor for the increase in incidence of tonsillar cancer. International Journal of Cancer. 2006;119(11):2620–2623. PubMed

Genden EM, Sambur IM, de Almeida JR, Posner M, Rinaldo A, Rodrigo JP, et al. Human papillomavirus and oropharyngeal squamous cell carcinoma: what the clinician should know. European Archives of Oto-Rhino-Laryngology. 2013;270(2):405–416. PubMed

Näsman A, Attner P, Hammarstedt L, et al. Incidence of human papillomavirus (HPV) positive tonsillar carcinoma in Stockholm, Sweden: an epidemic of viral-induced carcinoma? International Journal of Cancer. 2009;125(2):362–366. PubMed

Chaturvedi AK, Engels EA, Anderson WF, Gillison ML. Incidence trends for human papillomavirus-related and -unrelated oral squamous cell carcinomas in the United States. Journal of Clinical Oncology. 2008;26(4):612–619. PubMed

Smith EM, Ritchie JM, Summersgill KF, et al. Age, sexual behavior and human papillomavirus infection in oral cavity and oropharyngeal cancers. International Journal of Cancer. 2004;108(5):766–772. PubMed

Sivasithamparam J, Visk CA, Cohen EE, King AC. Modifiable risk behaviors in patients with head and neck cancer. Cancer. 2013;119(13):2419–2426. PubMed PMC

Rotnaglova E, Tachezy R, Salakova M, Prochazka B, Kosl'abova E, Vesela E, et al. HPV involvement in tonsillar cancer: prognostic significance and clinically relevant markers. International Journal of Cancer Journal International Du Cancer. 2011;129:101–110. PubMed

Attner P, Du J, Näsman A, et al. Human papillomavirus and survival in patients with base of tongue cancer. International Journal of Cancer. 2011;128(12):2892–2897. PubMed

Plummer M, Schiffman M, Castle PE, Maucort-Boulch D, Wheeler CM. A 2-year prospective study of human papillomavirus persistence among women with a cytological diagnosis of atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesion. Journal of Infectious Diseases. 2007;195(11):1582–1589. PubMed

Saussez S, Duray A, Demoulin S, Hubert P, Delvenne P. Immune suppression in head and neck cancers: a review. Clinical and Developmental Immunology. 2010;2010:15 pages.701657 PubMed PMC

Hoffmann TK, Dworacki G, Tsukihiro T, et al. Spontaneous apoptosis of circulating T lymphocytes in patients with head and neck cancer and its clinical importance. Clinical Cancer Research. 2002;8(8):2553–2562. PubMed

Whiteside TL. Immunobiology of head and neck cancer. Cancer and Metastasis Reviews. 2005;24(1):95–105. PubMed

Renoux VM, Bisig B, Langers I, et al. Human papillomavirus entry into NK cells requires CD16 expression and triggers cytotoxic activity and cytokine secretion. European Journal of Immunology. 2011;41(11):3240–3252. PubMed

Textor S, Dürst M, Jansen L, et al. Activating NK cell receptor ligands are differentially expressed during progression to cervical cancer. International Journal of Cancer. 2008;123(10):2343–2353. PubMed

Wulff S, Pries R, Börngen K, Trenkle T, Wollenberg B. Decreased levels of circulating regulatory NK cells in patients with head and neck cancer throughout all tumor stages. Anticancer Research. 2009;29(8):3053–3057. PubMed

Accomando WP, Wiencke JK, Houseman EA, Butler RA, Zheng S, Nelson HH, et al. Decreased NK cells in patients with head and neck cancer determined in archival DNA. Clinical Cancer Research. 2012;18:6147–6154. PubMed PMC

Molling JW, Langius JAE, Langendijk JA, et al. Low levels of circulating invariant natural killer T cells predict poor clinical outcome in patients with head and neck squamous cell carcinoma. Journal of Clinical Oncology. 2007;25(7):862–868. PubMed

Liyanage UK, Moore TT, Joo H-G, et al. Prevalence of regulatory T cells is increased in peripheral blood and tumor microenvironment of patients with pancreas or breast adenocarcinoma. Journal of Immunology. 2002;169(5):2756–2761. PubMed

Salama P, Phillips M, Grieu F, et al. Tumor-infiltrating FOXP3+ T regulatory cells show strong prognostic significance in colorectal cancer. Journal of Clinical Oncology. 2009;27(2):186–192. PubMed

Karagoz B, Bilgi O, Gümüs M, et al. CD8+CD28− cells and CD4+CD25+ regulatory T cells in the peripheral blood of advanced stage lung cancer patients. Medical Oncology. 2010;27(1):29–33. PubMed

Fu J, Zhang Z, Zhou L, Qi Z, Xing S, Lv J, et al. Impairment of CD4(+) cytotoxic T cells predicts poor survival and high recurrence rates in patients with hepatocellular carcinoma. Hepatology. 2013;58(1):139–149. PubMed

Ladányi A, Somlai B, Gilde K, Fejös Z, Gaudi I, Tímár J. T-cell activation marker expression on tumor-infiltrating lymphocytes as prognostic factor in cutaneous malignant melanoma. Clinical Cancer Research. 2004;10(2):521–530. PubMed

Bhairavabhotla RK, Verma V, Tongaonkar H, Shastri S, Dinshaw K, Chiplunkar S. Role of IL-10 in immune suppression in cervical cancer. Indian Journal of Biochemistry and Biophysics. 2007;44(5):350–356. PubMed

Lau K-M, Cheng SH, Lo KW, et al. Increase in circulating Foxp3+ CD4+ CD25high regulatory T cells in nasopharyngeal carcinoma patients. British Journal of Cancer. 2007;96(4):617–622. PubMed PMC

Schaefer C, Kim GG, Albers A, Hoermann K, Myers EN, Whiteside TL. Characteristics of CD4+CD25+ regulatory T cells in the peripheral circulation of patients with head and neck cancer. British Journal of Cancer. 2005;92(5):913–920. PubMed PMC

Strauss L, Bergmann C, Gooding W, Johnson JT, Whiteside TL. The frequency and suppressor function of CD4+CD25highFoxp3+ T cells in the circulation of patients with squamous cell carcinoma of the head and neck. Clinical Cancer Research. 2007;13(21):6301–6311. PubMed

Boucek J, Mrkvan T, Chovanec M, et al. Regulatory T cells and their prognostic value for patients with squamous cell carcinoma of the head and neck. Journal of Cellular and Molecular Medicine. 2010;14(1-2):426–433. PubMed PMC

Badoual C, Hans S, Rodriguez J, et al. Prognostic value of tumor-infiltrating CD4+ T-cell subpopulations in head and neck cancers. Clinical Cancer Research. 2006;12(2):465–472. PubMed

Zhang Y-L, Li J, Mo H-Y, et al. Different subsets of tumor infiltrating lymphocytes correlate with NPC progression in different ways. Molecular Cancer. 2010;9, article 4 PubMed PMC

Wamsom D, Light E, Thomas D, et al. Infiltrating lymphocytes and human papillomavirus-16-associated oropharyngeal cancer. Laryngoscope. 2012;122(1):121–127. PubMed PMC

Näsman A, Romanitan M, Nordfors C, Grun N, Johansson H, Hammarstedt L, et al. Tumor infiltrating CD8+ and Foxp3+ lymphocytes correlate to clinical outcome and human papillomavirus (HPV) status in tonsillar cancer. PloS ONE. 2012;7e38711 PubMed PMC

Aarstad HJ, Heimdal J-H, Klementsen B, Olofsson J, Ulvestad E. Presence of activated T lymphocytes in peripheral blood of head and neck squamous cell carcinoma patients predicts impaired prognosis. Acta Oto-Laryngologica. 2006;126(12):1326–1333. PubMed

Green VL, Michno A, Stafford ND, Greenman J. Increased prevalence of tumour infiltrating immune cells in oropharyngeal tumours in comparison to other subsites: relationship to peripheral immunity. Cancer Immunology, Immunotherapy. 2013;62:863–873. PubMed PMC

Koslabova E, Hamsikova E, Salakova M, Klozar J, Foltynova E, Salkova E, et al. Markers of HPV infection and survival in patients with head and neck tumours. International Journal of Cancer. 2013;133(8):1832–1839. PubMed

Klozar J, Kratochvil V, Salakova M, et al. HPV status and regional metastasis in the prognosis of oral and oropharyngeal cancer. European Archives of Oto-Rhino-Laryngology. 2008;265(1):S75–S82. PubMed

Tachezy R, Klozar J, Saláková M, et al. HPV and other risk factors of oral cavity/oropharyngeal cancer in the Czech Republic. Oral Diseases. 2005;11(3):181–185. PubMed

van den Brule AJC, Pol R, Fransen-Daalmeijer N, Schouls LM, Meijer CJLM, Snijders PJF. GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes. Journal of Clinical Microbiology. 2002;40(3):779–787. PubMed PMC

Gravitt PE, Peyton C, Wheeler C, Apple R, Higuchi R, Shah KV. Reproducibility of HPV 16 and HPV 18 viral load quantitation using TaqMan real-time PCR assays. Journal of Virological Methods. 2003;112(1-2):23–33. PubMed

Smeets SJ, Hesselink AT, Speel E-JM, et al. A novel algorithm for reliable detection of human papillomavirus in paraffin embedded head and neck cancer specimen. International Journal of Cancer. 2007;121(11):2465–2472. PubMed

Raghavan S, Quiding-Jädierbrink M. Regulatory T cells in gastrointestinal tumors. Expert Review of Gastroenterology and Hepatology. 2011;5(4):489–501. PubMed

Ladoire S, Martin F, Ghiringhelli F. Prognostic role of FOXP3+ regulatory T cells infiltrating human carcinomas: the paradox of colorectal cancer. Cancer Immunology, Immunotherapy. 2011;60(7):909–918. PubMed PMC

Ke X, Wang J, Li J, Chen IH, Wang H, Yang X-F. Roles of CD4+CD25high FOXP3+ Tregs in lymphomas and tumors are complex. Frontiers in Bioscience. 2008;13(10):3986–4001. PubMed

Álvaro T, Lejeune M, Salvadó MT, et al. Outcome in Hodgkin’s lymphoma can be predicted from the presence of accompanying cytotoxic and regulatory T cells. Clinical Cancer Research. 2005;11(4):1467–1473. PubMed

Roncador G, Brown PJ, Maestre L, et al. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. European Journal of Immunology. 2005;35(6):1681–1691. PubMed

Sakaguchi S, Sakaguchi N, Asano M, Itoh M, Toda M. Pillars article: immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J. Immunol. 1995. Journal of Immunology. 2011;186:3808–3821. PubMed

Smith EM, Rubenstein LM, Haugen TH, Pawlita M, Turek LP. Complex etiology underlies risk and survival in head and neck cancer human papillomavirus, tobacco, and alcohol: a case for multifactor disease. Journal of Oncology. 2012;2012:9 pages.571862 PubMed PMC

Hoffmann M, Tribius S, Quabius ES, et al. HPV DNA, E6*I-mRNA expression and p16INK4A immunohistochemistry in head and neck cancer—how valid is p16INK4A as surrogate marker? Cancer Letters. 2012;323(1):88–96. PubMed

Ames E, Murphy WJ. Advantages and clinical applications of natural killer cells in cancer immunotherapy. Cancer Immunology, Immunotherapy. 2014;63(1):21–28. PubMed PMC

PD1+CD8+ Cells Are an Independent Prognostic Marker in Patients with Head and Neck Cancer

The Head and Neck Squamous Cell Carcinoma Microenvironment as a Potential Target for Cancer Therapy