Approximately 15% of advanced head and neck squamous cell carcinomas (HNSCC) respond to anti-PD-(L)1 monotherapies. Tumor PD-L1 expression and human papillomavirus (HPV) status have been proposed as biomarkers to identify patients likely to benefit from these treatments. We aimed to understand the potential immune effects of HPV in HNSCC and to characterize additional potentially targetable immune-regulatory pathways in primary, treatment-naïve tumors. CD3, CD4, CD8, CD20, CD68, FoxP3, PD-1, PD-L2, LAG-3, IDO-1, and GITR cell densities were determined in 27 HNSCC specimens. IHC for PD-L1 assessed percentage of positive tumor cells and immune cells separately or as a combined positive score (CPS), and whether PD-L1 was expressed in an adaptive or constitutive pattern (i.e., PD-L1+ tumor cells juxtaposed to TILs or in the absence of TILs, respectively). HPV testing with p16 IHC was confirmed by HPV genotyping. When compared to HPV(−) tumors (n = 14), HPV+ tumors (n = 13) contained significantly higher densities of CD3+, CD4+, CD8+, CD20+, and PD-1+ cells (P < 0.02), and there was a trend towards increased density of FoxP3 + cells. PD-L1 expression patterns did not vary by tumor viral status, suggesting possible heterogeneous mechanisms driving constitutive vs adaptive PD-L1 expression patterns in HNSCC. IDO-1 expression was abundant (> 500 IDO-1+ cells/mm2 in 17/27 specimens) and was found on tumor cells as well as immune cells in 12/27 (44%) cases (range 5–80% tumor cells+). Notably, the studied markers varied on a per-patient basis and were not always related to the degree of T cell infiltration. These findings may inform therapeutic co-targeting strategies and raise consideration for a personalized treatment approach.
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The datasets generated during the current study are available from the corresponding author on reasonable request.
Mandal R, Senbabaoglu Y, Desrichard A, Havel JJ, Dalin MG, Riaz N et al (2016) The head and neck cancer immune landscape and its immunotherapeutic implications. JCI Insight 1:e89829
Chaturvedi AK, Engels EA, Pfeiffer RM, Hernandez BY, Xiao W, Kim E et al (2011) Human papillomavirus and rising oropharyngeal cancer incidence in the United States. J Clin Oncol 29:4294–4301
Ferris RL, Blumenschein G Jr, Fayette J, Guigay J, Colevas AD, Licitra L et al (2016) Nivolumab for recurrent squamous-cell carcinoma of the head and neck. N Engl J Med 375:1856–1867
Chow LQ, Haddad R, Gupta S, Mahipal A, Mehra R, Tahara M et al (2016) 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 34:3838–3845
Cohen EEW, Soulieres D, Le Tourneau C, Dinis J, Licitra L, Ahn MJ et al (2019) Pembrolizumab versus methotrexate, docetaxel, or cetuximab for recurrent or metastatic head-and-neck squamous cell carcinoma (KEYNOTE-040): a randomised, open-label, phase 3 study. Lancet 393:156–167
Burtness B, Harrington KJ, Greil R, Soulières D, Tahara M, de Castro, Jr G et al (2019) Pembrolizumab alone or with chemotherapy versus cetuximab with chemotherapy for recurrent or metastatic squamous cell carcinoma of the head and neck (KEYNOTE-048): a randomised, open-label, phase 3 study. Lancet 394:1915–1928
Seiwert TY, Zuo Z, Keck MK, Khattri A, Pedamallu CS, Stricker T et al (2015) Integrative and comparative genomic analysis of HPV-positive and HPV-negative head and neck squamous cell carcinomas. Clin Cancer Res 21:632–641
Saada-Bouzid E, Defaucheux C, Karabajakian A, Coloma VP, Servois V, Paoletti X et al (2017) Hyperprogression during anti-PD-1/PD-L1 therapy in patients with recurrent and/or metastatic head and neck squamous cell carcinoma. Ann Oncol 28:1605–1611
Zou W, Wolchok JD, Chen L (2016) PD-L1 (B7–H1) and PD-1 pathway blockade for cancer therapy: mechanisms, response biomarkers, and combinations. Sci Transl Med. 8:328rv324
Koyama S, Akbay EA, Li YY, Herter-Sprie GS, Buczkowski KA, Richards WG et al (2016) Adaptive resistance to therapeutic PD-1 blockade is associated with upregulation of alternative immune checkpoints. Nat Commun 7:10501
Yearley JH, Gibson C, Yu N, Moon C, Murphy E, Juco J et al (2017) PD-L2 expression in human tumors: relevance to anti-PD-1 therapy in cancer. Clin Cancer Res 23:3158–3167
Appah EO, Ballard BR, Izban MG, Jolin C, Lammers PE, Parrish DD Jr, Marshall DR (2018) A rapidly growing human papillomavirus-positive oral tongue squamous cell carcinoma in a 21-year old female: a case report. Oncol Lett 15:7702–7706
Henneman R, Van Monsjou HS, Verhagen CVM, Van Velthuysen MF, Haar NTT, Osse EM et al (2015) Incidence changes of human papillomavirus in oropharyngeal squamous cell carcinoma and effects on survival in the Netherlands Cancer Institute, 1980–2009. Anticancer Res 35:4015–4022
Sunshine JC, Nguyen P, Kaunitz G, Cottrell TR, Berry S, Esandrio J et al (2017) PD-L1 expression in melanoma: a quantitative immunohistochemical antibody comparison. Clin Cancer Res. 23:4938–4944
Lipson EJ, Lilo MT, Ogurtsova A, Esandrio J, Xu H, Brothers P et al (2017) Basal cell carcinoma: PD-L1/PD-1 checkpoint expression and tumor regression after PD-1 blockade. J Immunother Cancer 5:23
Yanik EL, Kaunitz GJ, Cottrell TR, Succaria F, McMiller TL, Ascierto ML et al (2017) Association of HIV status with local immune response to anal squamous cell carcinoma: implications for immunotherapy. JAMA Oncol. 3:974–978
Duffield AS, Ascierto ML, Anders RA, Taube JM, Meeker AK, Chen S et al (2017) Th17 immune microenvironment in Epstein-Barr virus-negative Hodgkin lymphoma: implications for immunotherapy. Blood Adv 1:1324–1334
Rimm DL, Han G, Taube JM, Yi ES, Bridge JA, Flieder DB et al (2017) A prospective, multi-institutional, pathologist-based assessment of 4 immunohistochemistry assays for PD-L1 expression in non-small cell lung cancer. JAMA Oncol 3(8):1051–1058
Hirsch FR, McElhinny A, Stanforth D, Ranger-Moore J, Jansson M, Kulangara K et al (2017) PD-L1 Immunohistochemistry assays for lung cancer: results from phase 1 of the blueprint PD-L1 IHC assay comparison project. J Thorac Oncol 12(2):208–222
Gaule P, Smithy JW, Toki M, Rehman J, Patell-Socha F, Cougot D et al (2017) A quantitative comparison of antibodies to programmed cell death 1 ligand 1. JAMA Oncol 3(2):256–259
Westra WH (2015) The pathology of HPV-related head and neck cancer: implications for the diagnostic pathologist. Semin Diagn Pathol 32:42–53
Taube JM, Anders RA, Young GD, Xu H, Sharma R, McMiller TL et al (2012) Colocalization of inflammatory response with B7–h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape. Sci Transl Med. 4:127ra137
Lyford-Pike S, Peng S, Young GD, Taube JM, Westra WH, Akpeng B et al (2013) Evidence for a role of the PD-1:PD-L1 pathway in immune resistance of HPV-associated head and neck squamous cell carcinoma. Cancer Res 73:1733–1741
Ang KK, Harris J, Wheeler R, Weber R, Rosenthal DI, Nguyen-Tân PF et al (2010) Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med 363:24–35
Taube JM, Galon J, Sholl LM, Rodig SJ, Cottrell TR, Giraldo NA et al (2018) Implications of the tumor immune microenvironment for staging and therapeutics. Mod Pathol 31:214–234
Giraldo NA, Nguyen P, Engle EL, Kaunitz GJ, Cottrell TR, Berry S et al (2018) Multidimensional, quantitative assessment of PD-1/PD-L1 expression in patients with Merkel cell carcinoma and association with response to pembrolizumab. J Immunother Cancer 6:99
Pardoll DM (2012) The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12:252–264
Steuer CE, Griffith CC, Nannapaneni S, Patel MR, Liu Y, Magliocca KR et al (2018) A correlative analysis of PD-L1, PD-1, PD-L2, EGFR, HER2, and HER3 expression in oropharyngeal squamous cell carcinoma. Mol Cancer Ther 17:710–716
Munn DH (2011) Indoleamine 2,3-dioxygenase, Tregs and cancer. Curr Med Chem 18:2240–2246
Schaer DA, Budhu S, Liu C, Bryson C, Malandro N, Cohen A et al (2013) GITR pathway activation abrogates tumor immune suppression through loss of regulatory T cell lineage stability. Cancer Immunol Res 1:320–331
Taube JM, Akturk G, Angelo M, Engle EL, Gnjatic S, Greenbaum S et al (2020) The Society for Immunotherapy in Cancer statement on best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) staining and validation. J Immunother Cancer 8:e000155
Luke JJ, Azad NS, Edwards R, Huang SMA, Comprelli A, Monga M et al (2018) Phase 1, open-label, adaptive biomarker trial that informs the evolution of combination immuno-oncology (IO) therapies (ADVISE), a precision IO approach to personalized medicine. J Clin Oncol. 36(15 suppl):TPS3101
The authors would like to acknowledge Drs. Robin Edwards and Darren Locke (Bristol-Myers Squibb) for helpful discussions and provision of the PD-L2 antibody.
This work was supported by the Bristol-Myers Squibb (PK, JH, SLT, JMT); National Cancer Institute R01 CA142779 (SLT, JMT); NIH T32 CA193145 (JES); and the Johns Hopkins Bloomberg-Kimmel Institute for Cancer Immunotherapy.
Conflict of interest
J. M. Taube reports consulting/advisory board for BMS, Merck, AstraZeneca, and Compugen; research funding through Bristol Myers Squibb; and reagents and machine loan from Akoya Biosciences. S. L. Topalian reports stock and other ownership interests in Aduro Biotech, DNAtrix, Dracen Pharmaceuticals, Dragonfly Therapeutics, Ervaxx, Five Prime Therapeutics, Potenza Therapeutics, RAPT, Tizona Therapeutics, Trieza Therapeutics, and WindMIL; a consulting or advisory role in Amgen, DNAtrix, Dragonfly Therapeutics, Dynavax, Ervaxx, Five Prime Therapeutics, Immunocore, Immunomic Therapeutics, Janssen Pharmaceuticals, MedImmune/AstraZeneca, Merck, RAPT, and WindMIL; research grants from Bristol Myers Squibb and Compugen; patents, royalties, and/or other intellectual property through her institution with Aduro Biotech, Arbor Pharmaceuticals, Bristol Myers Squibb, Immunomic Therapeutics, NexImmune, and WindMIL; and travel, accommodations, and expenses from Bristol-Myers Squibb and Five Prime Therapeutics. P. Kvistborg is a consultant for Neon Therapeutics and Personalis and a recipient of grant/research support from Bristol-Myers Squibb and Merck. J. Haanen: NKI received financial compensation for advisory role of J. Haanen with AZ, Amgen, Bayer, BMS, Celsius Therapeutics, MSD, Merck Serono, Pfizer, Roche/Genentech, Neon Therapeutics, lmmunocore, Seattle Genetics, Novartis, GSK. Also, NKI received research grants through J. Haanen from BMS, MSD, Novartis, and Neon Therapeutics. J. Stein reports consulting (uncompensated) for AstraZeneca. No potential conflicts of interest were disclosed by the other authors.
The local Ethics Committees of the Netherlands Cancer Institute (NKI) and Johns Hopkins University (JHU) approved this study. The study is retrospective in nature and all procedures performed were part of routine care.
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Succaria, F., Kvistborg, P., Stein, J.E. et al. Characterization of the tumor immune microenvironment in human papillomavirus-positive and -negative head and neck squamous cell carcinomas. Cancer Immunol Immunother 70, 1227–1237 (2021). https://doi.org/10.1007/s00262-020-02747-w