Medical Oncology

, 36:21 | Cite as

Characteristics and impact of programmed death-ligand 1 expression, CD8+ tumor-infiltrating lymphocytes, and p16 status in head and neck squamous cell carcinoma

  • Nuttapong NgamphaiboonEmail author
  • Teeranuch Chureemas
  • Teerada Siripoon
  • Lalida Arsa
  • Narumol Trachu
  • Chuleeporn Jiarpinitnun
  • Poompis Pattaranutaporn
  • Ekaphop Sirachainan
  • Noppadol Larbcharoensub
Original Paper



No predictive biomarker of immune checkpoint inhibitors in head and neck squamous cell carcinoma (HNSCC) has been well established. The impact of programmed death-ligand 1 (PD-L1) expression, CD8+ tumor-infiltrating lymphocytes (TILs), and p16 status in HNSCC is unclear and may vary according to ethnicity.


HNSCC patients treated between 2007 and 2013 were reviewed. Archival tissues were retrieved for PD-L1, CD8+ TILs, and p16 analyses. PD-L1 expression was evaluated by using the validated SP142 assay on the VENTANA platform. CD8+ TILs were defined by using semiquantitative scoring.


A total of 203 patients were analyzed. PD-L1 expression was observed in 80% of patients and was significantly associated with older age (P < 0.001). A high CD8+ TIL score (≥ 6) was significantly associated with never-smoking (P = 0.020), oral cavity cancer (P < 0.001), and stage M0 at presentation (P = 0.025). The p16 status was positive in 12% of patients. Patients with a high TIL score had a significantly longer OS (P = 0.032). Patients with PD-L1 expression of 1–49% and ≥ 50% were associated with a significantly shorter OS compared with those with PD-L1 < 1% (P = 0.027 and P = 0.011, respectively). Multivariate analysis showed that PD-L1 ≥ 50% was significantly associated with a poor OS. (HR 2.98 [95% CI 1.2–7.39]; P = 0.019.)


A high prevalence of PD-L1 expression was observed in HNSCC using the validated SP142 assay. PD-L1 expression was associated with older age, while highly PD-L1 expression (≥ 50%) was an independent prognostic factor for poor OS in anti-PD1/PD-L1 untreated HNSCC patients.


Programmed death ligand-1 CD8+ tumor infiltrating lymphocytes P16 Human papillomavirus Head and neck squamous cell carcinoma 



This work was supported by a grant from the Ramathibodi Cancer Center Grant. N. Ngamphaiboon received funding from the Research University Network (RUN), and the Thailand Research Fund (TRF), and the Research Development Grant from the Ramathibodi Hospital. The authors thank Ms. Dollapas Punpanich for statistical analysis.

Compliance with ethical standards

Conflict of interest

All authors declare no conflict of interest.

Informed consent

The Ramathibodi Ethic Committee approved a waiver of consent for this study as a retrospective chart review. Archrival tissues used in this study were considered as a leftover specimen. The research involves no more than minimal risk to the subject and is not adversely affect the rights and welfare of the subjects. All patient identifications were protected according to the GCP guideline and not published in the manuscript. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Supplementary material

12032_2018_1241_MOESM1_ESM.docx (67 kb)
Supplementary material 1 (DOCX 67 KB)


  1. 1.
    Ling DC, Bakkenist CJ, Ferris RL, Clump DA. Role of immunotherapy in head and neck cancer. Semin Radiat Oncol. 2018;28(1):12–6. Scholar
  2. 2.
    Solomon B, Young RJ, Rischin D. Head and neck squamous cell carcinoma: genomics and emerging biomarkers for immunomodulatory cancer treatments. Semin Cancer Biol. 2018. Scholar
  3. 3.
    Qi X, Jia B, Zhao X, Yu D. Advances in T-cell checkpoint immunotherapy for head and neck squamous cell carcinoma. OncoTargets Therapy. 2017;10:5745–54. Scholar
  4. 4.
    Seiwert TY, Burtness B, Mehra R, Weiss J, Berger R, Eder JP, et al. Safety and clinical activity of pembrolizumab for treatment of recurrent or metastatic squamous cell carcinoma of the head and neck (KEYNOTE-012): an open-label, multicentre, phase 1b trial. Lancet Oncol. 2016;17(7):956–65. Scholar
  5. 5.
    Chow LQM, Haddad R, Gupta S, Mahipal A, Mehra R, Tahara M, et al. Antitumor activity of pembrolizumab in biomarker-unselected patients with recurrent and/or metastatic head and neck squamous cell carcinoma: results from the phase Ib KEYNOTE-012 expansion cohort. J Clin Oncol. 2016;34(32):3838–45. Scholar
  6. 6.
    Ferris RL, Blumenschein G Jr, Fayette J, Guigay J, Colevas AD, Licitra L, et al. Nivolumab for recurrent squamous-cell carcinoma of the head and neck. N Engl J Med. 2016;375(19):1856–67. Scholar
  7. 7.
    Bauml J, Seiwert TY, Pfister DG, Worden F, Liu SV, Gilbert J, et al. Pembrolizumab for platinum- and cetuximab-refractory head and neck cancer: results from a single-arm, phase II study. J Clin Oncol. 2017;35(14):1542–9. Scholar
  8. 8.
    Ferris R, Blumenschein G Jr, Fayette J, Guigay J, Colevas AD, Licitra L, et al. Nivolumab vs investigator’s choice in recurrent or metastatic squamous cell carcinoma of the head and neck: 2-year long-term survival update of CheckMate 141 with analyses by tumor PD-L1 expression. Oral Oncol. 2018;81:45–51.CrossRefGoogle Scholar
  9. 9.
    Larkin J, Hodi FS, Wolchok JD. Combined nivolumab and ipilimumab or monotherapy in untreated melanoma. N Engl J Med. 2015;373(13):1270–1. Scholar
  10. 10.
    Robert C, Long GV, Brady B, Dutriaux C, Maio M, Mortier L, et al. Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med. 2015;372(4):320–30. Scholar
  11. 11.
    Herbst RS, Soria JC, Kowanetz M, Fine GD, Hamid O, Gordon MS, et al. Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients. Nature. 2014;515(7528):563–7. Scholar
  12. 12.
    Heineman TE, Widman A, Kuan EC, St John M. The genetic landscape of programmed death ligand-1 (PD-L1) alterations in head and neck cancer. Laryngoscope Investig Otolaryngol. 2017;2(3):99–103. Scholar
  13. 13.
    Wollenberg B. Cancer Immunology and HPV. Recent results in cancer research Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer. 2017;206:243–8. Scholar
  14. 14.
    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. Scholar
  15. 15.
    Balermpas P, Rodel F, Rodel C, Krause M, Linge A, Lohaus F, et al. 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. 2016;138(1):171–81. Scholar
  16. 16.
    Hansen AR, Siu LL. PD-L1 testing in cancer: challenges in companion diagnostic development. JAMA Oncol. 2016;2(1):15–6. Scholar
  17. 17.
    Bahleda R, Braiteh FS, Balmanoukian AS, Braña I, Hodi FS, Garbo L, et al. Long-term safety and clinical outcomes of atezolizumab in head and neck cancer: phase ia trial results. Ann Oncol. 2017. Scholar
  18. 18.
    De Meulenaere A, Vermassen T, Aspeslagh S, Deron P, Duprez F, Laukens D, et al. Tumor PD-L1 status and CD8(+) tumor-infiltrating T cells: markers of improved prognosis in oropharyngeal cancer. Oncotarget. 2017;8(46):80443–52. Scholar
  19. 19.
    Chow LQM, Mehra R, Haddad RI, Mahipal A, Weiss J, Berger R, et al. Biomarkers and response to pembrolizumab (pembro) in recurrent/metastatic head and neck squamous cell carcinoma (R/M HNSCC). JCO. 2016;34(15):suppl.6010.CrossRefGoogle Scholar
  20. 20.
    Soulieres D, Cohen EE, Le Tourneau C, Dinis J, Licitra L, Ahn MJ, et al. Updated survival results of the KEYNOTE-040 study of pembrolizumab vs standard-of-care chemotherapy for recurrent or metastatic head and neck squamous cell carcinoma. In: AACR; Chicago, IL; 2018.Google Scholar
  21. 21.
    Segal NH, Ou SHI, Balmanoukian AS, Massarelli E, Brahmer JR, Weiss J, et al. Updated safety and efficacy of durvalumab (MEDI4736), an anti-PD-L 1 antibody, in patients from a squamous cell carcinoma of the head and neck (SCCHN) expansion cohort. Ann Oncol. 2016. Scholar
  22. 22.
    Pai S, Cohen EE, Lin D, Fountzilas G, Kim ES, Mehlhorn H, et al. RetroSpective cohort stUdy of PD-L1 expression in REcurrent and/or MEtastatic squamous cell carcinoma of the head and neck (SUPREME-HN). Ann Oncol. 2017;28(suppl_5):6040.CrossRefGoogle Scholar
  23. 23.
    Ono T, Azuma K, Kawahara A, Sasada T, Hattori S, Sato F, et al. Association between PD-L1 expression combined with tumor-infiltrating lymphocytes and the prognosis of patients with advanced hypopharyngeal squamous cell carcinoma. Oncotarget. 2017;8(54):92699–714. Scholar
  24. 24.
    Oguejiofor K, Galletta-Williams H, Dovedi SJ, Roberts DL, Stern PL, West CM. Distinct patterns of infiltrating CD8+ T cells in HPV+ and CD68 macrophages in HPV− oropharyngeal squamous cell carcinomas are associated with better clinical outcome but PD-L1 expression is not prognostic. Oncotarget. 2017;8(9):14416–27. Scholar
  25. 25.
    Muller T, Braun M, Dietrich D, Aktekin S, Hoft S, Kristiansen G, et al. PD-L1: a novel prognostic biomarker in head and neck squamous cell carcinoma. Oncotarget. 2017;8(32):52889–900. Scholar
  26. 26.
    Hirsch FR, McElhinny A, Stanforth D, Ranger-Moore J, Jansson M, Kulangara K, et al. PD-L1 immunohistochemistry assays for lung cancer: results from phase 1 of the blueprint PD-L1 IHC assay comparison project. J Thorac Oncol. 2017;12(2):208–22. Scholar
  27. 27.
    Rimm DL, Han G, Taube JM, Yi ES, Bridge JA, Flieder DB, et al. A prospective multi-institutional, pathologist-based assessment of 4 immunohistochemistry assays for PD-L1 expression in non-small cell lung cancer. JAMA Oncol. 2017;3(8):1051–8. Scholar
  28. 28.
    Badoual C, Hans S, Merillon N, Van Ryswick C, Ravel P, Benhamouda N, et al. PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. Cancer Res. 2013;73(1):128–38. Scholar
  29. 29.
    Stokes M, Wang R, Wildsmith S, Secrier M, Angell HK, Barker C, et al. Relationship between PD-L1 expression and survival in head and neck squamous cell carcinoma (HNSCC) patients (pts). Ann. Oncol. 2017;28(suppl_5):v372–v394.CrossRefGoogle Scholar
  30. 30.
    Hendry S, Salgado R, Gevaert T, Russell PA, John T, Thapa B, et al. Assessing tumor-infiltrating lymphocytes in solid tumors: a practical review for pathologists and proposal for a standardized method from the international immuno-oncology biomarkers working group: part 2: TILs in melanoma, gastrointestinal tract carcinomas, non-small cell lung carcinoma and mesothelioma, endometrial and ovarian carcinomas, squamous cell carcinoma of the head and neck, genitourinary carcinomas, and primary brain tumors. Adv Anat Pathol. 2017;24(6):311–35. Scholar
  31. 31.
    Sritippho T, Pongsiriwet S, Lertprasertsuke N, Buddhachat K, Sastraruji T, Iamaroon A. p16-a possible surrogate marker for high-risk human papillomaviruses in oral cancer? Asian Pac J Cancer Prev. 2016;17(8):4049–57.PubMedGoogle Scholar
  32. 32.
    Phusingha P, Ekalaksananan T, Vatanasapt P, Loyha K, Promthet S, Kongyingyoes B, et al. Human papillomavirus (HPV) infection in a case-control study of oral squamous cell carcinoma and its increasing trend in northeastern Thailand. J Med Virol. 2017;89(6):1096–101. Scholar
  33. 33.
    Dogan V, Rieckmann T, Munscher A, Busch CJ. Current studies of immunotherapy in head and neck cancer. Clin Otolaryngol. 2018;43(1):13–21. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Nuttapong Ngamphaiboon
    • 1
    Email author
  • Teeranuch Chureemas
    • 1
  • Teerada Siripoon
    • 1
  • Lalida Arsa
    • 2
  • Narumol Trachu
    • 3
  • Chuleeporn Jiarpinitnun
    • 4
  • Poompis Pattaranutaporn
    • 4
  • Ekaphop Sirachainan
    • 1
  • Noppadol Larbcharoensub
    • 2
  1. 1.Division of Medical Oncology, Department of Medicine, Faculty of Medicine Ramathibodi HospitalMahidol UniversityBangkokThailand
  2. 2.Department of Pathology, Faculty of Medicine Ramathibodi HospitalMahidol UniversityBangkokThailand
  3. 3.Ramathibodi Research Center, Faculty of Medicine Ramathibodi HospitalMahidol UniversityBangkokThailand
  4. 4.Division of Radiation Oncology, Department of Radiology, Faculty of Medicine Ramathibodi HospitalMahidol UniversityBangkokThailand

Personalised recommendations