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Characterization of the tumor immune microenvironment in human papillomavirus-positive and -negative head and neck squamous cell carcinomas


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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Data availability

The datasets generated during the current study are available from the corresponding author on reasonable request.


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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.

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Correspondence to Janis M. Taube.

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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.

Ethical approval

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).

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