Cellular and Molecular Biology

Intratumor heterogeneity of PD-L1 expression in head and neck squamous cell carcinoma

Article metrics


Intratumor heterogeneity may contribute to the ambiguous clinical results on PD-L1 status as a predictor for immunotherapy response in patients with HNSCC. This decreases the utility of PD-L1 expression from single tumour biopsies as a predictive biomarker. In this prospective study, intratumor heterogeneity of PD-L1 expression in HNSCC was investigated with both Tumour Proportion Score (TPS) and Combined Positive Score (CPS). Thirty-three whole surgical specimens from 28 patients with HNSCC were included. PD-L1 expression in six random core biopsies from each surgical specimen was used to assess the concordance between multiple biopsies and the negative predictive value of a single negative core biopsy. With 1% cut off, 36% of the specimens were concordant with TPS and 52% with CPS. With a 50% cut-off value the concordance was 70% with TPS and 55% with CPS. Defining a tumour as positive if just a single-one of the biopsies was positive, the negative predictive value (NPV) of a single negative core biopsy was 38.9 and 0% (1% cut off), and 79.9% and 62.8% (50% cut off) for TPS and CPS, respectively. In conclusion, PD-L1 positivity varies markedly within the tumour, both with TPS and CPS, challenging the utility of this biomarker.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1
Fig. 2


  1. 1.

    Network TCGA. Comprehensive genomic characterization of head and neck squamous cell carcinomas. Nature 517, 576–582 (2015).

  2. 2.

    Pignon, J.-P., le Maître, A., Maillard, E. & Bourhis, J. Meta-analysis of chemotherapy in head and neck cancer (MACH-NC): an update on 93 randomised trials and 17,346 patients. Radiother Oncol. 92, 4–14 (2009).

  3. 3.

    Vermorken, J. B., Mesia, R., Rivera, F., Remenar, E., Kawecki, A., Rottey, S. et al. Platinum-based chemotherapy plus cetuximab in head and neck cancer. N. Engl. J. Med. 359, 1116–1127 (2008).

  4. 4.

    Szturz, P. & Vermorken, J. B. Immunotherapy in head and neck cancer: aiming at EXTREME precision. BMC Med. 15, 110 (2017).

  5. 5.

    Ferris, R. L., Blumenschein, G., Fayette, J., Guigay, J., Colevas, A. D., 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. 81, 45–51 (2018).

  6. 6.

    Ferris, R. L., Blumenschein, G., Fayette, J., Guigay, J., Colevas, A. D., Licitra, L. et al. Nivolumab for recurrent squamous-cell carcinoma of the head and neck. N. Engl. J. Med. 375, 1856–1867 (2016).

  7. 7.

    Bauml, J., Seiwert, T. Y., Pfister, D. G., Worden, F., Liu, S. V., 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. 35, 1542–1549 (2017).

  8. 8.

    Cohen E. E., Harrington K. J., Le Tourneau C., Dinis J., Licitra L., Ahn M.-J. et al. LBA45_PRPembrolizumab (pembro) vs standard of care (SOC) for recurrent or metastatic head and neck squamous cell carcinoma (R/M HNSCC): Phase 3 KEYNOTE-040 trial. Ann. Oncol. 2017; 28. https://doi.org/10.1093/annonc/mdx440.040.

  9. 9.

    Harrington K. J., Ferris R. L., Blumenschein G., Colevas D., Fayette J., Licitra L. et al. Nivolumab versus standard, single-agent therapy of investigator’s choice in recurrent or metastatic squamous cell carcinoma of the head and neck (CheckMate 141): health-related quality-of-life results from a randomised, phase 3 trial. Lancet Oncol. 18, 1104–1115 (2017).

  10. 10.

    Mehra, R., Seiwert, T. Y., Gupta, S., Weiss, J., Gluck, I., Eder, J. P. et al. Efficacy and safety of pembrolizumab in recurrent/metastatic head and neck squamous cell carcinoma: pooled analyses after long-term follow-up in KEYNOTE-012. Br. J. Cancer 119, 153–159 (2018).

  11. 11.

    Chow, L. Q. M., 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. 34, 3838–3845 (2016).

  12. 12.

    Antonia S. J., Villegas A., Daniel D., Vicente D., Murakami S., Hui R. et al. Overall Survival with Durvalumab after Chemoradiotherapy in Stage III NSCLC. N. Engl. J. Med. 379, 2342–2350 (2018).

  13. 13.

    Hirsch, F. R., 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. 12, 208–222 (2017).

  14. 14.

    Rimm, D. L., Han, G., Taube, J. M., Yi, E. S., Bridge, J. A., Flieder, D. B. 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. 3, 1051 (2017).

  15. 15.

    Wang C., Hahn E., Slodkowska E., Eskander A., Enepekides D., Higgins K. et al. Reproducibility of PD-L1 immunohistochemistry interpretation across various types of genitourinary and head/neck carcinomas, antibody clones, and tissue types. Hum Pathol 82, 131–139 (2018). https://doi.org/10.1016/j.humpath.2018.07.024.

  16. 16.

    Rehman, J. A., Han, G., Carvajal-Hausdorf, D. E., Wasserman, B. E., Pelekanou, V., Mani, N. L. et al. Quantitative and pathologist-read comparison of the heterogeneity of programmed death-ligand 1 (PD-L1) expression in non-small cell lung cancer. Mod Pathol 30, 340–349 (2017).

  17. 17.

    McLaughlin, J., Han, G., Schalper, K. A., Carvajal-Hausdorf, D., Pelekanou, V., Rehman, J. et al. Quantitative assessment of the heterogeneity of PD-L1 expression in non-small-cell lung cancer. JAMA Oncol. 2, 46–54 (2016).

  18. 18.

    Bhaijee, F. & Anders, R. A. PD-L1 expression as a predictive biomarker. JAMA Oncol. 2, 54 (2016).

Download references


J.H.R. has received a grant from Rigshospitalets research committee and from The Danish Cancer Society. S.M.B. is supported in part by grant no. P30 CA 134274-04 from the NCI and from the KFJ Foundation.

Author information

J.H.R., I.R.V., S.M.B. and J.F. contributed to the design and conduct. J.H.R., G.L., H.H.J., B.M.F. and I.W., contributed to the acquisition of the data. K.H., L.S., C.A.K., C.v.B. and I.W. contributed to the analysis, and J.H.R., I.V.R. and S.M.B. performed statistical support. All authors contributed substantial to the interpretation of data and were involved in drafting and revising the manuscript. All authors approved the final version of the manuscript.

Correspondence to Jacob H. Rasmussen.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval

The study was approved by the Regional Committee on Health Research Ethics (The Capital Region of Denmark), approval number H-16049387, and study was performed in accordance with the Declaration of Helsinki.

Data availability

The dataset used and analysed during the current study is available from the corresponding author on reasonable request.


This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Rasmussen, J.H., Lelkaitis, G., Håkansson, K. et al. Intratumor heterogeneity of PD-L1 expression in head and neck squamous cell carcinoma. Br J Cancer 120, 1003–1006 (2019) doi:10.1038/s41416-019-0449-y

Download citation

Further reading