Immune cells and cytolytic activity within the tumor microenvironment are being intensively studied. Through transcriptome profiling, immune cell enumeration using the xCell tool and cytolytic activity quantification according to granzyme A (GZMA) and perforin (PRF1) mRNA levels, we investigated immunoreactivity in tumor and/or tumor-free tongue tissue samples from 31 patients with squamous cell carcinoma of the oral tongue and 14 healthy individuals (control tongue tissues). We found significantly altered immune cell compositions (p < 0.001) and elevated cytolytic activity (p < 0.001) in tumor compared to tumor-free samples, and altered infiltration of a subset of immune cells (e.g. CD8+ T cells, p < 0.01) as well as increased cytolytic activity (p < 0.001) in tumor-free compared to control samples. Controlling for patient age at diagnosis and tumor stage, Cox regression analysis showed that high cytolytic activity in tumor-free samples associated with improved disease-free survival (hazard ratio= 4.20, 95% CI = 1.09-16.20, p = 0.037). However, the degree of cytolytic activity in tumor samples did not provide prognostic information. Taken together, our results show the presence of cancer-related immune responses in clinically tumor-free tongue in patients with squamous cell carcinoma of the oral tongue. Measuring cytolytic activity in tumor-free tongue samples contralateral to tumor might thus be an effective approach to predict clinical outcome.

Department of Clinical Sciences ENT Umeå University Umeå Sweden

Department of Medical Biosciences Pathology Umeå University Umeå Sweden

Institute of Molecular Genetics University Paris 7 St Louis Hospital Paris France

Regional Centre for Applied Molecular Oncology Masaryk Memorial Cancer Institute Brno Czech Republic

See more in PubMed

Binnewies M, Roberts EW, Kersten K, et al Understanding the tumor immune microenvironment (TIME) for effective therapy. Nat Med 2018; 24: 541–550. PubMed PMC

Baumeister SH, Freeman GJ, Dranoff G, et al Coinhibitory pathways in immunotherapy for cancer. Annu Rev Immunol 2016; 34: 539–573. PubMed

Hackl H, Charoentong P, Finotello F, et al Computational genomics tools for dissecting tumour‐immune cell interactions. Nat Rev Genet 2016; 17: 441–458. PubMed

Charoentong P, Finotello F, Angelova M, et al Pan‐cancer immunogenomic analyses reveal genotype‐immunophenotype relationships and predictors of response to checkpoint blockade. Cell Rep 2017; 18: 248–262. PubMed

Thorsson V, Gibbs DL, Brown SD, et al The immune landscape of cancer. Immunity 2018; 48: 812–830 e814. PubMed PMC

Gentles AJ, Newman AM, Liu CL, et al The prognostic landscape of genes and infiltrating immune cells across human cancers. Nat Med 2015; 21: 938–945. PubMed PMC

Galon J, Mlecnik B, Bindea G, et al Towards the introduction of the 'Immunoscore' in the classification of malignant tumours. J Pathol 2014; 232: 199–209. PubMed PMC

Pages F, Mlecnik B, Marliot F, et al International validation of the consensus Immunoscore for the classification of colon cancer: a prognostic and accuracy study. Lancet 2018; 391: 2128–2139. PubMed

Finotello F, Trajanoski Z. Quantifying tumor‐infiltrating immune cells from transcriptomics data. Cancer Immunol Immunother 2018; 67: 1031–1040. PubMed PMC

Aran D, Hu Z, Butte AJ. xCell: digitally portraying the tissue cellular heterogeneity landscape. Genome Biol 2017; 18: 220. PubMed PMC

Aran D, Camarda R, Odegaard J, et al Comprehensive analysis of normal adjacent to tumor transcriptomes. Nat Commun 2017; 8: 1077. PubMed PMC

Martinez‐Lostao L, Anel A, Pardo J. How do cytotoxic lymphocytes kill cancer cells? Clin Cancer Res 2015; 21: 5047–5056. PubMed

Voskoboinik I, Whisstock JC, Trapani JA. Perforin and granzymes: function, dysfunction and human pathology. Nat Rev Immunol 2015; 15: 388–400. PubMed

Rooney MS, Shukla SA, Wu CJ, et al Molecular and genetic properties of tumors associated with local immune cytolytic activity. Cell 2015; 160: 48–61. PubMed PMC

Bray F, Ferlay J, Soerjomataram I, et al Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018; 68: 394–424. PubMed

Leemans CR, Snijders PJF, Brakenhoff RH. The molecular landscape of head and neck cancer. Nat Rev Cancer 2018; 18: 269–282. PubMed

Solomon B, Young RJ, Rischin D. Head and neck squamous cell carcinoma: genomics and emerging biomarkers for immunomodulatory cancer treatments. Semin Cancer Biol 2018; 52: 228–240. PubMed

Boldrup L, Coates PJ, Wahlgren M, et al Subsite‐based alterations in miR‐21, miR‐125b, and miR‐203 in squamous cell carcinoma of the oral cavity and correlation to important target proteins. J Carcinog 2012; 11: 18. PubMed PMC

Cancer Genome Atlas N. Comprehensive genomic characterization of head and neck squamous cell carcinomas. Nature 2015; 517: 576–582. PubMed PMC

Boldrup L, Gu X, Coates PJ, et al Gene expression changes in tumor free tongue tissue adjacent to tongue squamous cell carcinoma. Oncotarget 2017; 8: 19389–19402. PubMed PMC

Gu X, Boldrup L, Coates PJ, et al Epigenetic regulation of OAS2 shows disease‐specific DNA methylation profiles at individual CpG sites. Sci Rep 2016; 6: 32579. PubMed PMC

Lundqvist L, Stenlund H, Laurell G, et al The importance of stromal inflammation in squamous cell carcinoma of the tongue. J Oral Pathol Med 2012; 41: 379–383. PubMed

Sgaramella N, Coates PJ, Strindlund K, et al Expression of p16 in squamous cell carcinoma of the mobile tongue is independent of HPV infection despite presence of the HPV‐receptor syndecan‐1. Br J Cancer 2015; 113: 321–326. PubMed PMC

Wilms T, Khan G, Coates PJ, et al No evidence for the presence of Epstein‐Barr virus in squamous cell carcinoma of the mobile tongue. PLoS One 2017; 12: e0184201. PubMed PMC

Shi W, Oshlack A, Smyth GK. Optimizing the noise versus bias trade‐off for Illumina whole genome expression BeadChips. Nucleic Acids Res 2010; 38: e204. PubMed PMC

Narayanan S, Kawaguchi T, Yan L, et al Cytolytic activity score to assess anticancer immunity in colorectal Cancer. Ann Surg Oncol 2018; 25: 2323–2331. PubMed PMC

Slaughter DP, Southwick HW, Smejkal W. Field cancerization in oral stratified squamous epithelium; clinical implications of multicentric origin. Cancer 1953; 6: 963–968. PubMed

Leemans CR, Braakhuis BJ, Brakenhoff RH. The molecular biology of head and neck cancer. Nat Rev Cancer 2011; 11: 9–22. PubMed

Curtius K, Wright NA, Graham TA. An evolutionary perspective on field cancerization. Nat Rev Cancer 2018; 18: 19–32. PubMed

Lochhead P, Chan AT, Nishihara R, et al Etiologic field effect: reappraisal of the field effect concept in cancer predisposition and progression. Mod Pathol 2015; 28: 14–29. PubMed PMC

Dotto GP. Multifocal epithelial tumors and field cancerization: stroma as a primary determinant. J Clin Invest 2014; 124: 1446–1453. PubMed PMC

Lohavanichbutr P, Houck J, Doody DR, et al Gene expression in uninvolved oral mucosa of OSCC patients facilitates identification of markers predictive of OSCC outcomes. PLoS One 2012; 7: e46575. PubMed PMC

Reis PP, Waldron L, Perez‐Ordonez B, et al A gene signature in histologically normal surgical margins is predictive of oral carcinoma recurrence. BMC Cancer 2011; 11: 437. PubMed PMC

Upregulation of Apoptosis Related Genes in Clinically Normal Tongue Contralateral to Squamous Cell Carcinoma of the Oral Tongue, an Effort to Maintain Tissue Homeostasis

Transfer-RNA-Derived Fragments Are Potential Prognostic Factors in Patients with Squamous Cell Carcinoma of the Head and Neck

Downregulation of TAP1 in Tumor-Free Tongue Contralateral to Squamous Cell Carcinoma of the Oral Tongue, an Indicator of Better Survival