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The repertoire of tumor-infiltrating lymphocytes within the microenvironment of oral squamous cell carcinoma reveals immune dysfunction

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Abstract

Background

The role of tumor-infiltrating lymphocytes (TILs) in the immune remodeling of tumor microenvironments (TME) in oral squamous cell carcinoma (OSCC) remains controversial. In this study, we pursued a comprehensive characterization of the repertoire of TILs and then analyzed its clinical significance and potential prognostic value.

Methods

Fresh tumor tissue samples and peripheral blood from 83 OSCC patients were collected to comprehensively characterize the phenotypes and frequencies of TILs by flow cytometry. Archived paraffin-embedded tissues derived from 159 OSCC patients were analyzed by immunohistochemistry to further assess the TIL repertoire. The clinical significance of TILs and their potential prognostic value were further analyzed.

Results

A series of unique features of TILs were observed. IL-17 was highly expressed in betel nut chewers, and CD20 was abundantly expressed in patients who did not drink alcohol; high expression of CD138, PD-L1, and Foxp3 was associated with poor prognosis. The Th17/Treg ratio was an independent prognostic factor for patient survival with greater predictive accuracy for overall survival.

Conclusions

Our results suggest an antigen-driven immune response; however, the immune dysfunction within the microenvironment in OSCC and the Th17/Treg balance may play important roles in the modulation of antitumor immunity.

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Abbreviations

AUC:

Area under curve

Bregs:

Regulatory B cells

CI:

Confidence interval

HNSCC:

Head and neck squamous cell carcinoma

HPFs:

High-power fields.

HRs:

Hazard ratios

OSCC:

Oral squamous cell carcinoma

ROC:

Receiver operating characteristic

Tc17:

T cytotoxic 17

TCR-Vβ:

T-cell receptor β chain variable region

Th17:

T helper 17

Tim-3:

T-cell immunoglobulin and mucin-domain containing-3

References

  1. Sasahira T, Kirita T, Kuniyasu H (2014) Update of molecular pathobiology in oral cancer: a review. Int J Clin Oncol 19:431–436

    Article  CAS  PubMed  Google Scholar 

  2. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108. https://doi.org/10.3322/caac.21262

    Article  PubMed  Google Scholar 

  3. Beynon RA, Lang S, Schimansky S et al (2018) Tobacco smoking and alcohol drinking at diagnosis of head and neck cancer and all-cause mortality: results from head and neck 5000, a prospective observational cohort of people with head and neck cancer. Int J Cancer 143:1114–1127. https://doi.org/10.1002/ijc.31416

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dhull AK, Atri R, Dhankhar R, Chauhan AK, Kaushal V (2018) Major risk factors in head and neck cancer: a retrospective analysis of 12-year experiences. World J Oncol 9:80–84. https://doi.org/10.14740/wjon1104w

    Article  PubMed  PubMed Central  Google Scholar 

  5. Madathil SA, Rousseau MC, Wynant W, Schlecht NF, Netuveli G, Franco EL, Nicolau B (2016) Nonlinear association between betel quid chewing and oral cancer: implications for prevention. Oral Oncol 60:25–31

    Article  CAS  PubMed  Google Scholar 

  6. Bloebaum M, Poort L, Böckmann R, Kessler P (2014) Survival after curative surgical treatment for primary oral squamous cell carcinoma. J Craniomaxillofac Surg 42:1572–1576

    Article  CAS  PubMed  Google Scholar 

  7. Rogers SN, Brown JS, Woolgar JA, Derek L, Patrick M, Shaw RJ, David S, Douglas E, David V (2009) Survival following primary surgery for oral cancer. Oral Oncol 45:201–211

    Article  PubMed  Google Scholar 

  8. Cor Hj L, Stefan S, Sabine VS et al (2013) Treatment of metastatic renal cell carcinoma with CAIX CAR-engineered T cells: clinical evaluation and management of on-target toxicity. Mol Therap J Am Soc Gene Therap 21:904–912

    Article  Google Scholar 

  9. Morgan RA, Yang JC, Mio K, Dudley ME, Laurencot CM, Rosenberg SA (2010) Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol Ther 18:843–851

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Tang H, Qiao J, Fu YX (2016) Immunotherapy and tumor microenvironment. Cancer Lett 370:85–90. https://doi.org/10.1016/j.canlet.2015.10.009

    Article  CAS  PubMed  Google Scholar 

  11. Hui L, Chen Y (2015) Tumor microenvironment: Sanctuary of the devil. Cancer Lett 368:7–13. https://doi.org/10.1016/j.canlet.2015.07.039

    Article  CAS  PubMed  Google Scholar 

  12. Balermpas P, Rödel F, Rödel C et al (2015) CD8+ tumour-infiltrating lymphocytes in relation to HPV status and clinical outcome in patients with head and neck cancer after postoperative chemoradiotherapy: a multicentre study of the German cancer consortium radiation oncology group (DKTK-ROG). Int J Cancer 138:171–181

    Article  PubMed  Google Scholar 

  13. Han S, Zhang C, Li Q, Dong J, Liu Y, Huang Y, Jiang T, Wu A (2014) Tumour-infiltrating CD4(+) and CD8(+) lymphocytes as predictors of clinical outcome in glioma. Br J Cancer 110:2560–2568. https://doi.org/10.1038/bjc.2014.162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Yoshioka T, Miyamoto M, Cho Y et al (2008) Infiltrating regulatory T cell numbers is not a factor to predict patient's survival in oesophageal squamous cell carcinoma. Br J Cancer 98:1258–1263. https://doi.org/10.1038/sj.bjc.6604294

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Müller S (2017) Update from the 4th Edition of the World Health Organization of head and neck tumours: tumours of the oral cavity and mobile tongue. Head Neck Pathol 11:33–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Doescher J, Veit JA, Hoffmann TK (2017) The 8th edition of the AJCC cancer staging manual. HNO 1–5

  17. Sherwood A, Emerson R, Scherer D et al (2013) Tumor-infiltrating lymphocytes in colorectal tumors display a diversity of T cell receptor sequences that differ from the T cells in adjacent mucosal tissue. Cancer Immunol Immunotherap CII 62:1453–1461

    Article  CAS  Google Scholar 

  18. Gubin MM, Artyomov MN, Mardis ER, Schreiber RD (2015) Tumor neoantigens: building a framework for personalized cancer immunotherapy. J Clin Investig 125:3413–3421

    Article  PubMed  PubMed Central  Google Scholar 

  19. Ward JP, Gubin MM, Schreiber RD (2016) The role of neoantigens in naturally occurring and therapeutically induced immune responses to cancer. Adv Immunol 130:25–74. https://doi.org/10.1016/bs.ai.2016.01.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Jiang P, Gu S, Pan D et al (2018) Signatures of T cell dysfunction and exclusion predict cancer immunotherapy response. Nat Med 24:1550–1558. https://doi.org/10.1038/s41591-018-0136-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Joyce JA, Fearon DT (2015) T cell exclusion, immune privilege, and the tumor microenvironment. Science 348:74

    Article  CAS  PubMed  Google Scholar 

  22. Angelosanto JM, Blackburn SD, Crawford A, Wherry EJ (2012) Progressive loss of memory T cell potential and commitment to exhaustion during chronic viral infection. J Virol 86:8161–8170

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Brooks DG, McGavern DB, Oldstone MBA (2006) Reprogramming of antiviral T cells prevents inactivation and restores T cell activity during persistent viral infection. J Clin Investig 116:1675

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Wherry EJ, Kurachi M (2015) Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol 15:486–499

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Grivennikov SI, Greten FR, Karin M (2010) Immunity, inflammation, and cancer. Cell 140:883–899. https://doi.org/10.1016/j.cell.2010.01.025

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Littman DR, Rudensky AY (2010) Th17 and regulatory T cells in mediating and restraining inflammation. Cell 140:845–858. https://doi.org/10.1016/j.cell.2010.02.021

    Article  CAS  PubMed  Google Scholar 

  27. Mizoguchi A, Bhan AK (2006) A case for regulatory B cells. J Immunol 176:705–710

    Article  CAS  PubMed  Google Scholar 

  28. Mizoguchi A, Mizoguchi E, Takedatsu H, Blumberg RS, Bhan AK (2002) Chronic intestinal inflammatory condition generates IL-10-producing regulatory B cell subset characterized by CD1d upregulation. Immunity 16:219–230

    Article  CAS  PubMed  Google Scholar 

  29. Fillatreau S, Sweenie CH, Mcgeachy MJ, Gray D, Anderton SM (2002) B cells regulate autoimmunity by provision of IL-10. Nat Immunol 3:944–950

    Article  CAS  PubMed  Google Scholar 

  30. Chen J, He Q, Liu J, Xiao Y, Xiao C, Chen K, Xie D, Zhang X (2018) CD8+ tumor-infiltrating lymphocytes as a novel prognostic biomarker in lung sarcomatoid carcinoma, a rare subtype of lung cancer. Cancer Manag Res 10:3505–3511. https://doi.org/10.2147/CMAR.S169074

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Ye SL, Li XY, Zhao K, Feng T (2017) High expression of CD8 predicts favorable prognosis in patients with lung adenocarcinoma: a cohort study. Medicine 96:e6472. https://doi.org/10.1097/MD.0000000000006472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Balermpas P, Michel Y, Wagenblast J, Seitz O, Weiss C, Rödel F, Rödel C, Fokas E (2014) Tumour-infiltrating lymphocytes predict response to definitive chemoradiotherapy in head and neck cancer. Br J Cancer 110:501–509

    Article  CAS  PubMed  Google Scholar 

  33. Fang J, Li X, Ma D et al (2017) Prognostic significance of tumor infiltrating immune cells in oral squamous cell carcinoma. BMC Cancer 17:375. https://doi.org/10.1186/s12885-017-3317-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Cecilia N, Nathalie G, Nikolaos T et al (2013) CD8+ and CD4+ tumour infiltrating lymphocytes in relation to human papillomavirus status and clinical outcome in tonsillar and base of tongue squamous cell carcinoma. Eur J Cancer 49:2522–2530

    Article  Google Scholar 

  35. Wolf GT, Chepeha DB, Bellile E, Nguyen A, Thomas D, McHugh J, University of Michigan H, Neck SP (2015) Tumor infiltrating lymphocytes (TIL) and prognosis in oral cavity squamous carcinoma: a preliminary study. Oral Oncol 51:90–95. https://doi.org/10.1016/j.oraloncology.2014.09.006

    Article  CAS  Google Scholar 

  36. Jackute J, Zemaitis M, Pranys D, Sitkauskiene B, Miliauskas S, Bajoriunas V, Lavinskiene S, Sakalauskas R (2015) The prognostic influence of tumor infiltrating Foxp3+CD4+, CD4+ and CD8+ T cells in resected non-small cell lung cancer. J Inflamm 12:63

    Article  Google Scholar 

  37. Liu H, Zhang T, Ye J, Li H, Huang J, Li X, Wu B, Huang X, Hou J (2012) Tumor-infiltrating lymphocytes predict response to chemotherapy in patients with advance non-small cell lung cancer. Cancer Immunol Immunotherap CII 61:1849–1856. https://doi.org/10.1007/s00262-012-1231-7

    Article  CAS  Google Scholar 

  38. Zom GG, Khan S, Britten CM et al (2014) Efficient induction of antitumor immunity by synthetic toll-like receptor ligand-peptide conjugates. Cancer Immunol Res 2:756–764

    Article  CAS  PubMed  Google Scholar 

  39. Zamarron BF, Chen WJ (2011) Dual roles of immune cells and their factors in cancer development and progression. Int J Biol Sci 7:651–658

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Sato E, Olson SH, Ahn J et al (2005) Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. Proc Natl Acad Sci USA 102:18538–18543

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Martin-Orozco N, Muranski P, Chung Y et al (2009) T helper 17 cells promote cytotoxic T cell activation in tumor immunity. Immunity 31:787–798

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Kryczek I, Banerjee M, Cheng P et al (2009) Phenotype, distribution, generation, and functional and clinical relevance of Th17 cells in the human tumor environments. Blood 114:1141–1149. https://doi.org/10.1182/blood-2009-03-208249

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Sfanos KS, Bruno TC, Maris CH, Xu L, Thoburn CJ, DeMarzo AM, Meeker AK, Isaacs WB, Drake CG (2008) Phenotypic analysis of prostate-infiltrating lymphocytes reveals TH17 and Treg skewing. Clin Cancer Res 14:3254–3261. https://doi.org/10.1158/1078-0432.CCR-07-5164

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Asadzadeh Z, Mohammadi H, Safarzadeh E, Hemmatzadeh M, Mahdian-Shakib A, Jadidi-Niaragh F, Azizi G, Baradaran B (2017) The paradox of Th17 cell functions in tumor immunity. Cell Immunol 322:15–25. https://doi.org/10.1016/j.cellimm.2017.10.015

    Article  CAS  PubMed  Google Scholar 

  45. Chen X, Wan J, Liu J, Xie W, Diao X, Xu J, Zhu B, Chen Z (2010) Increased IL-17-producing cells correlate with poor survival and lymphangiogenesis in NSCLC patients. Lung Cancer 69:348–354. https://doi.org/10.1016/j.lungcan.2009.11.013

    Article  PubMed  Google Scholar 

  46. Guery L, Hugues S (2015) Th17 cell plasticity and functions in cancer immunity. Biomed Res Int 2015:314620. https://doi.org/10.1155/2015/314620

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Di GP, Gerlini G, Caporale R, Sestini S, Brandani P, Urso C, Pimpinelli N, Borgognoni L (2018) T regulatory cells mediate immunosuppresion by adenosine in peripheral blood, sentinel lymph node and TILs from melanoma patients. Cancer Lett 417:124–130

    Article  Google Scholar 

  48. Herpen CMLV, Voort RVD, Laak JAVD et al (2010) Intratumoral rhIL-12 administration in head and neck squamous cell carcinoma patients induces B cell activation. Int J Cancer 123:2354–2361

    Article  Google Scholar 

  49. Jie-Yi S, Qiang G, Zhi-Chao W et al (2013) Margin-infiltrating CD20+ B cells display an atypical memory phenotype and correlate with favorable prognosis in hepatocellular carcinoma. Clin Cancer Res 19:5994–6005

    Article  Google Scholar 

  50. Marcus S, Daniel BH, Christian VTR et al (2008) The humoral immune system has a key prognostic impact in node-negative breast cancer. Can Res 68:5405–5413

    Article  Google Scholar 

  51. Taghavi N, Mohsenifar Z, Baghban AA, Arjomandkhah A (2018) CD20+ Tumor infiltrating B lymphocyte in oral squamous cell carcinoma: correlation with clinicopathologic characteristics and heat shock protein 70 expression. Pathol Res Int 2018:4810751. https://doi.org/10.1155/2018/4810751

    Article  Google Scholar 

  52. Berntsson J, Nodin B, Eberhard J, Micke P, Jirstrom K (2016) Prognostic impact of tumour-infiltrating B cells and plasma cells in colorectal cancer. Int J Cancer 139:1129–1139. https://doi.org/10.1002/ijc.30138

    Article  CAS  PubMed  Google Scholar 

  53. Al-Shibli KI, Tom D, Samer AS, Magnus P, Bremnes RM, Lill-Tove B (2008) Prognostic effect of epithelial and stromal lymphocyte infiltration in non-small cell lung cancer. Clin Cancer Res 14:5220

    Article  CAS  PubMed  Google Scholar 

  54. Wouters MC, Nelson BH (2018) Prognostic significance of tumor-infiltrating B cells and plasma cells in human cancer. Clin Cancer Res. https://doi.org/10.1158/1078-0432.CCR-18-1481

    Article  PubMed  Google Scholar 

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Funding

This work was supported by the National Natural Science Foundation of China (81501110), Hunan Province Natural Science Foundation (2016JJ4093), and China Postdoctoral Science Foundation (2016M592454).

Author information

Authors and Affiliations

  1. Research Institution of Stomatology, Xiangya Stomatological Hospital and School of Stomatology, Central South University, 72 Xiangya Road, Changsha, 410008, Hunan, People’s Republic of China

    Hongzhi Quan, Zhongyan Shan, Ziyi Liu, Sixuan Liu, Liu Yang, Xiaodan Fang, Kun Li, Baisheng Wang, Zhiyuan Deng, Yanjia Hu, Zhigang Yao, Junhui Huang & Zhangui Tang

  2. Department of Oral Maxillofacial Surgery, Xiangya Stomatological Hospital and School of Stomatology, Central South University, Changsha, 410008, Hunan, People’s Republic of China

    Hongzhi Quan, Zhongyan Shan, Ziyi Liu, Sixuan Liu, Liu Yang, Xiaodan Fang, Kun Li, Baisheng Wang, Zhiyuan Deng, Yanjia Hu & Zhangui Tang

  3. Department of Oral Pathology, Xiangya Stomatological Hospital and School of Stomatology, Central South University, Changsha, 410008, Hunan, People’s Republic of China

    Zhigang Yao & Junhui Huang

  4. Department of Head and Neck Surgery, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, 410008, Hunan, People’s Republic of China

    Jianjun Yu

  5. Center for Medical Genetics, School of Life Science, Central South University, Changsha, 410013, People’s Republic of China

    Kun Xia

  6. Department of Neurology, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, 410008, Hunan, People’s Republic of China

    Liangjuan Fang

  7. Key Laboratory of Hunan Province in Neurodegenerative Disorders, Central South University, Changsha, 410008, Hunan, People’s Republic of China

    Liangjuan Fang

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  1. Hongzhi Quan
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  2. Zhongyan Shan
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  3. Ziyi Liu
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  4. Sixuan Liu
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  5. Liu Yang
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  6. Xiaodan Fang
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  7. Kun Li
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  14. Kun Xia
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  15. Zhangui Tang
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Contributions

Conception and design: HZQ, ZYS, and LJF; Data acquisition: ZYS, ZYL, SXL, LY, XDF, BSW, and KL; Evaluation of pathology and IHC: ZYD, YJH, JHH, and ZGY; Data analysis and interpretation: HZQ, ZYS, LJF, ZYL, LY, JJY, KX, and TZT; Writing and review of manuscript: HZQ, ZYS, LJF, and ZGT.

Corresponding authors

Correspondence to Hongzhi Quan or Liangjuan Fang.

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Conflict of interest

None of the authors report any conflict of interest.

Ethical approval

This work was approved by the Ethics Committee of Xiangya Stomatological Hospital and School of Stomatology under the Reference Number 2016003 and was conducted in accordance with the principles of the Declaration of Helsinki.

Informed consent

Written informed consent to the use of specimens and clinical data for research purposes was obtained from the patients of the flow cytometry cohort before recruitment into the study. For the IHC cohort, the patient also signed consent to the use of their specimens and clinical data for research purposes before surgical resection.

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Quan, H., Shan, Z., Liu, Z. et al. The repertoire of tumor-infiltrating lymphocytes within the microenvironment of oral squamous cell carcinoma reveals immune dysfunction. Cancer Immunol Immunother 69, 465–476 (2020). https://doi.org/10.1007/s00262-020-02479-x

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