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Dynamic alterations of circulating T lymphocytes and the clinical response in patients with head and neck squamous cell carcinoma treated with nivolumab

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Abstract

Cancer immunotherapy using immune checkpoint inhibitors (ICIs) has been recognized as a novel therapeutic option for head and neck squamous cell carcinoma (HNSCC). However, only approximately 20–30% of patients with recurrent/metastatic (R/M) HNSCC benefit. Moreover, the mechanisms underlying the response to ICIs remain unclear. We investigated the proportion, activation status, and expression level of immune checkpoint molecules in circulating T cell subsets in R/M HNSCC patients treated with nivolumab using flow cytometry and mass cytometry, and then determined whether treatment response was associated with these values. We also assessed the changes in the frequency of tumor-associated antigens, MAGE-A4 and p53, -specific T cells prior to and after nivolumab treatment using the IFN-γ ELISPOT assay. The proportion of activated CD4+ and CD8+ TEMRA cells significantly increased in the disease-controlled patients but not in disease-progressed patients. As expected, the expression of PD-1 in T cells markedly decreased regardless of the therapeutic response. Meanwhile, T cell immunoglobulin mucin-3 expression on CD8+ T cells was significantly higher in patients with disease progression than in disease-controlled patients after treatment. The frequency of the tumor-associated antigens, MAGE-A4- and p53-specific T cells, was not correlated with clinical responses; however, in the disease-controlled patients, the frequency of MAGE-A4-specific T cells was significantly augmented. We concluded that in R/M HNSCC patients treated with nivolumab, circulating T cells show dynamic alterations depending on treatment efficacy. An analysis of the immunokinetics of circulating T cells could thus provide new insights into rational therapeutic strategies in cancer immunotherapy for HNSCC.

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Availability of data and material

The datasets used and/or analyzed in this study are available from the corresponding author upon reasonable request.

Abbreviations

CTLA-4:

Cytotoxic T-lymphocyte-associated protein 4

CyTOF:

Cytometry by time-of-flight

DAB:

3,3’-Diaminobenzidine

ELISPOT:

Enzyme-linked immunosorbent spot

FCS:

Fetal calf serum

Foxp3:

Forkhead box P3

HNSCC:

Head and neck squamous cell carcinoma

IFN:

Interferon

IHC:

Immunohistochemistry

ICI:

Immune checkpoint inhibitor

IL:

Interleukin

Lag-3:

Lymphocyte activation gene 3

MAGE-A4:

Melanoma-associated antigen-A4

MDS:

Multi-dimensional scaling

PBMC:

Peripheral blood mononuclear cell

PD-1:

Programmed cell death-1

PD-L1:

Programmed cell death-1 ligand 1

PHA:

Phytohemagglutinin

R/M:

Recurrent/metastatic

SFC:

Spot-forming-cells

TAA:

Tumor-associated antigen

TCM:

Central memory T cells

TEM:

Effector memory T cells

TEMRA:

Terminally differentiated effector memory T cells

Tim-3:

T cell immunoglobulin mucin-3

TN:

Naïve T cells

UMAP:

Uniform manifold approximation and projection

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Acknowledgements

We would like to thank Editage (www.editage.com) for English language editing.

Funding

This work was supported in part by a Grant-in-Aid for Scientific Research (Grant No. 20K18243 to Hideyuki T., No. 20K09747 to M.S., No. 19K18794 to S.I., No 19K18758 to I.M., and No. 20H03834 to K.C.) from the Ministry of Education, Culture, Sports, Science and Technology; a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Research Project for Sericultural Revolution), Japan; and Financial Support for the Promotion of University-Industry Collaborative Research Based on Regulatory Sciences at Gunma University.

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Author notes

  1. Hiroe Tada and Hideyuki Takahashi contributed equally to this study.

Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, 3-39-22, Showa-machi, Maebashi, Gunma, 3718511, Japan

    Hiroe Tada, Hideyuki Takahashi, Yurino Nagata, Miho Uchida, Masato Shino, Shota Ida, Ikko Mito, Toshiyuki Matsuyama & Kazuaki Chikamatsu

  2. Division of Molecular Science, Faculty of Science and Technology, Gunma University, Kiryu, Gunma, 376-8515, Japan

    Kanae Yamada & Shigeki Takeda

  3. Department of Pathology, Gunma University Graduate School of Medicine, Maebashi, Gunma, 371-8511, Japan

    Kei Masuda & Tetsunari Oyama

  4. Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, 305-8634, Japan

    Ken-Ichiro Tatematsu & Hideki Sezutsu

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  1. Hiroe Tada
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Contributions

Hideyuki T. and KC were involved in conception and design. Hiroe T., Hideyuki T., KY, KM, YN, MU, MS, SI, IM, TM, KT and HS were involved in sample preparation and data acquisition. Hiroe T., Hideyuki T., TO, ST and KC were involved in analysis and interpretation of data. Hiroe T. and KC were involved in writing of the manuscript. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Kazuaki Chikamatsu.

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The authors declare that they have no conflict interest.

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This study was approved by the Ethical Committee of Gunma University Hospital (HS2017-152).

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Written informed consent was obtained from each patient.

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Tada, H., Takahashi, H., Yamada, K. et al. Dynamic alterations of circulating T lymphocytes and the clinical response in patients with head and neck squamous cell carcinoma treated with nivolumab. Cancer Immunol Immunother 71, 851–863 (2022). https://doi.org/10.1007/s00262-021-03042-y

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