The immune system is important for elimination of cancer cells. Tumors including oral squamous cell carcinoma (OSCC) are capable of escaping detection by host immune cells through apoptotic depletion of tumor-infiltrating lymphocytes (TILs). Circulating peripheral blood lymphocytes (PBLs) and corresponding TILs of tumor specimen were evaluated before and after curative tumor resection (n = 30) compared with PBLs of controls (n = 87). PBLs were characterized for the total number of T cells (CD3+), T helper cells (Th, CD3+/CD4+), regulatory T cells (Treg, CD4+/CD25+/CD127low), cytotoxic T cells (Tc, CD3+/CD8+), activated T cells (CD3+/HLA-DR+), and natural killer (NK) cells (CD3−/CD16+/CD56+). In tumor tissue, the prevalence of CD3+, CD4+, and CD8+ TILs was assessed using immunohistochemistry, whereas the incidence of apoptosis was assessed using terminal deoxynucleotidyl transferase deoxyuridinetriphosphate nick-end labeling (TUNEL) assay. In PBLs of pretreated OSCC patients, a highly significant decrease in total number of T cells (p = 0.0001), Th cells (p < 0.0001), Treg cells (p < 0.0001), Tc cells (p < 0.0001), and NK cells (p = 0.0037) were found compared with controls. Decreased PBLs of OSCC patients were correlated with decreased numbers of corresponding TILs, which were associated with increased detection of apoptosis in the tumor tissue. Compared with the controls, the total number of T cells remained unchanged after surgery but the total number of NK cells significantly increased. Standardized immunophenotyping of OSCC may help to identify patients likely to benefit from cancer immunotherapy strategies and/or chemoradiation. Finally, future attempts to enhance an effective tumor-reactive immune response by immunotherapy or vaccination should be made by promoting tumor-specific Th and/or Tc cell/NK cell responses.
Oral squamous cell carcinoma Immunophenotyping Immunotherapy Cytotoxic T cells Regulatory T cells
Peripheral blood lymphocytes
Oral squamous cell carcinoma
T helper cells
Regulatory T cells
Cytotoxic T cells
Natural killer cells
This is a preview of subscription content, log in to check access.
We thank biovis’ Diagnostik MVZ especially Melanie Hügen and Martina Thümmler for the technical support.
Compliance with ethical standards
MG and OF conceived the study, performed the coordination, and drafted the manuscript. OF performed flow cytometric analysis. TB and AM analyzed histopathological specimen and carried out immunohistochemistry studies. MG and PT carried out the data collection and performed the statistical analyses. SR performed surgical treatment, after care of the patients, and drafted the manuscript. All authors read and approved the final manuscript.
Conflicts of interest
Consent to participate
Written informed consent to participate was obtained prospectively from all patients (Ethics Committee Tübingen, Germany, approval number: 562-2013BO2).
Badoual C, Sandoval F, Pere H, Hans S, Gey A, Merillon N, et al. Better understanding tumor-host interaction in head and neck cancer to improve the design and development of immunotherapeutic strategies. Head Neck. 2010;32(7):946–58. doi:10.1002/hed.21346.PubMedGoogle Scholar
Tanaka H, Yoshizawa H, Yamaguchi Y, Ito K, Kagamu H, Suzuki E, et al. Successful adoptive immunotherapy of murine poorly immunogenic tumor with specific effector cells generated from gene-modified tumor-primed lymph node cells. J Immunol. 1999;162(6):3574–82.PubMedGoogle Scholar
Igney FH, Krammer PH. Immune escape of tumors: apoptosis resistance and tumor counterattack. J Leukoc Biol. 2002;71(6):907–20.PubMedGoogle Scholar
Gastman BR, Atarshi Y, Reichert TE, Saito T, Balkir L, Rabinowich H, et al. Fas ligand is expressed on human squamous cell carcinomas of the head and neck, and it promotes apoptosis of T lymphocytes. Cancer Res. 1999;59(20):5356–64.PubMedGoogle Scholar
Kassouf N, Thornhill MH. Oral cancer cell lines can use multiple ligands, including Fas-L, TRAIL and TNF-alpha, to induce apoptosis in Jurkat T cells: possible mechanisms for immune escape by head and neck cancers. Oral Oncol. 2008;44(7):672–82. doi:10.1016/j.oraloncology.2007.08.013.CrossRefPubMedGoogle Scholar
Reichert TE, Strauss L, Wagner EM, Gooding W, Whiteside TL. Signaling abnormalities, apoptosis, and reduced proliferation of circulating and tumor-infiltrating lymphocytes in patients with oral carcinoma. Clin Cancer Res. 2002;8(10):3137–45.PubMedGoogle Scholar
Kim JW, Wieckowski E, Taylor DD, Reichert TE, Watkins S, Whiteside TL. Fas ligand-positive membranous vesicles isolated from sera of patients with oral cancer induce apoptosis of activated T lymphocytes. Clin Cancer Res. 2005;11(3):1010–20.PubMedGoogle Scholar
Hoffmann TK, Dworacki G, Tsukihiro T, Meidenbauer N, Gooding W, Johnson JT, et al. Spontaneous apoptosis of circulating T lymphocytes in patients with head and neck cancer and its clinical importance. Clin Cancer Res. 2002;8(8):2553–62.PubMedGoogle Scholar
Wolf GT, Hudson JL, Peterson KA, Miller HL, McClatchey KD. Lymphocyte subpopulations infiltrating squamous carcinomas of the head and neck: correlations with extent of tumor and prognosis. Otolaryngol Head Neck Surg. 1986;95(2):142–52.CrossRefPubMedGoogle Scholar
Wolf GT, Schmaltz S, Hudson J, Robson H, Stackhouse T, Peterson KA, et al. Alterations in T-lymphocyte subpopulations in patients with head and neck cancer. Correlations with prognosis. Arch Otolaryngol Head Neck Surg. 1987;113(11):1200–6.CrossRefPubMedGoogle Scholar
Balermpas P, Michel Y, Wagenblast J, Seitz O, Weiss C, Rodel F, et al. Tumour-infiltrating lymphocytes predict response to definitive chemoradiotherapy in head and neck cancer. Br J Cancer. 2014;110(2):501–9. doi:10.1038/bjc.2013.640.CrossRefPubMedGoogle Scholar
Gaur P, Qadir GA, Upadhyay S, Singh AK, Shukla NK, Das SN. Skewed immunological balance between Th17 (CD4(+)IL17A (+)) and Treg (CD4 (+)CD25 (+)FOXP3 (+)) cells in human oral squamous cell carcinoma. Cell Oncol (Dordr). 2012;35(5):335–43. doi:10.1007/s13402-012-0093-5.CrossRefGoogle Scholar
Gasparoto TH, de Souza Malaspina TS, Benevides L, de Melo EJ, Costa Jr MR, Damante JH, et al. Patients with oral squamous cell carcinoma are characterized by increased frequency of suppressive regulatory T cells in the blood and tumor microenvironment. Journal of the Formosan immunol, immunother: CII. 2010;59(6):819–28. doi:10.1007/s00262-009-0803-7.CrossRefGoogle Scholar
Drennan S, Stafford ND, Greenman J, Green VL. Increased frequency and suppressive activity of CD127(low/-) regulatory T cells in the peripheral circulation of patients with head and neck squamous cell carcinoma are associated with advanced stage and nodal involvement. Immunology. 2013;140(3):335–43. doi:10.1111/imm.12144.PubMedPubMedCentralGoogle Scholar
Rammensee HG, Weinschenk T, Gouttefangeas C, Stevanovic S. Towards patient-specific tumor antigen selection for vaccination. Immunol Rev. 2002;188:164–76.CrossRefPubMedGoogle Scholar
Singh-Jasuja H, Emmerich NP, Rammensee HG. The Tubingen approach: identification, selection, and validation of tumor-associated HLA peptides for cancer therapy. Cancer immunology, immunotherapy : CII. 2004;53(3):187–95. doi:10.1007/s00262-003-0480-x.CrossRefPubMedGoogle Scholar
Feyen O, Coy JF, Prasad V, Schierl R, Saenger J, Baum RP. EDIM-TKTL1 blood test: a noninvasive method to detect upregulated glucose metabolism in patients with malignancies. Future Oncol. 2012;8(10):1349–59. doi:10.2217/fon.12.98.CrossRefPubMedGoogle Scholar
Grimm M, Cetindis M, Lehmann M, Biegner T, Munz A, Teriete P, et al. Association of cancer metabolism-related proteins with oral carcinogenesis—indications for chemoprevention and metabolic sensitizing of oral squamous cell carcinoma? J Transl Med. 2014;12:208. doi:10.1186/1479-5876-12-208.CrossRefPubMedPubMedCentralGoogle Scholar
Zweig MH, Campbell G. Receiver-operating characteristic (ROC) plots: a fundamental evaluation tool in clinical medicine. Clin Chem. 1993;39(4):561–77.PubMedGoogle Scholar
Timar J, Ladanyi A, Forster-Horvath C, Lukits J, Dome B, Remenar E, et al. Neoadjuvant immunotherapy of oral squamous cell carcinoma modulates intratumoral CD4/CD8 ratio and tumor microenvironment: a multicenter phase II clinical trial. J Clin Oncol. 2005;23(15):3421–32. doi:10.1200/JCO.2005.06.005.CrossRefPubMedGoogle Scholar
Yeh CY, Lin CL, Chang MC, Chen HM, Kok SH, Chang SH, et al. Differences in oral habit and lymphocyte subpopulation affect malignant transformation of patients with oral precancer. J Formos Med Assoc=Taiwan yi zhi. 2015. doi:10.1016/j.jfma.2015.07.017.Google Scholar
Tabata T, Hazama S, Yoshino S, Oka M. Th2 subset dominance among peripheral blood T lymphocytes in patients with digestive cancers. Am J Surg. 1999;177(3):203–8.CrossRefPubMedGoogle Scholar
Young M. Immunological phenotypes of premalignant oral lesions and the immune shifts with the development of head and neck cancer. Austin J Otolaryngol. 2014;1(2):7.Google Scholar