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Characteristic differences in the abundance of tumor-infiltrating lymphocytes and intratumoral developing T cells in thymoma, with special reference to PD-1 expression

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Abstract

Purpose

Though programmed cell death-1 (PD-1) inhibitors mainly target tumor-infiltrating lymphocytes (TILs) expressing PD-1, developing T cells in thymus also express PD-1 in their process of maturation. To predict the therapeutic effect of PD-1 inhibitors for thymoma, it is necessary to clarify the proportions of TILs and intratumoral developing T cells.

Methods

The expressions of CD4, CD8, and PD-1 on T cells were analyzed by flow cytometry in 31 thymomas. The amount of T cell receptor excision circles (TRECs), which can be detected in newly formed naïve T cells in the thymus, was evaluated using sorted lymphocytes from thymomas by quantitative PCR. The expressions of granzyme B (GZMB) and lymphocyte activation gene-3 (LAG-3) in PD-1 + CD8 T cells were analyzed by image cytometry using multiplex immunohistochemistry.

Results

The PD-1 + rate in both CD4 and CD8 T cells was significantly higher in type AB/B1/B2 than in type A/B3 thymomas. The amounts of TRECs in CD4 and CD8 T cells were significantly higher in type AB/B1/B2 than in type A/B3 thymomas and comparable to normal thymus. PD-1 expression at each stage of T cell development of type AB/B1/B2 thymomas was comparable to that of normal thymus. Both the percentages and cell densities of PD-1 + CD8 T cells expressing GZMB or LAG-3, which are known to contain tumor-reactive T cells, were significantly lower in type AB/B1/B2 thymomas.

Conclusion

Most PD-1 + T cells in type AB/B1/B2 thymomas are intratumoral developing T cells and are not TILs.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ACTB:

Actin beta

CD:

Cluster of differentiation

DP:

Double-positive

GZMA:

Granzyme A

GZMB:

Granzyme B

I/O:

Immune-oncology

irAE:

Immune-related adverse event

LAG-3:

Lymphocyte activation gene-3

mIHC:

Multiplex immunohistochemistry

PBMC:

Peripheral blood mononuclear cell

PD-1:

Programmed death receptor-1

PD-L1:

Programmed death ligand 1

ROI:

Region of interest

SP:

Single-positive

TCR:

T cell receptor

TIL:

Tumor-infiltrating lymphocyte

TREC:

T cell receptor excision circle

WHO:

World Health Organization

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Acknowledgments

The authors would like to thank D. Kato (Japan Red Cross Kyoto Daini Hospital), Y. Ueshima (Japan Red Cross Kyoto Daiichi Hospital), and T. Ii (Ayabe City Hospital) for their important contribution of providing thymoma specimens in the present study.

Funding

This work was supported by a Grant-in-Aid for Scientific Research, Japan Society for Promotion of Science (16K10705), and a Grant for Lung Cancer Research, The Japan Lung Cancer Society. The funding agencies had no role in the present study.

Author information

Authors and Affiliations

  1. Division of Thoracic Surgery, Department of Surgery, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kamigyo-ku, Kyoto, 602-8566, Japan

    Tatsuo Furuya, Shunta Ishihara, Chiaki Nakazono, Satoru Okada, Masanori Shimomura & Masayoshi Inoue

  2. Lab of Immunology, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan

    Tatsuo Furuya, Kyoko Masuda & Hiroshi Kawamoto

  3. Department of Pathology and Applied Neurobiology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    Hiroshi Ogi, So Tando, Takeshi Yaoi & Kyoko Itoh

  4. SCREEN Holdings Co., Ltd, Kyoto, Japan

    Hiroshi Ogi & Saya Shibata

  5. Department of Pediatric Cardiovascular Surgery, Children’s Medical Center, Kyoto Prefectural University of Medicine, Kyoto, Japan

    Yoshinobu Maeda & Masaaki Yamagishi

Authors
  1. Tatsuo Furuya
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  2. Shunta Ishihara
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Contributions

TF and MI contributed to the study conception and design. Material preparation and data collection were performed by TF, SI, CN, YM, and MY. Data analysis was performed by TF, SI, HO, SS, ST, and TY. The first draft of the manuscript was written by TF. Review and editing were performed by KM, SO, MS, HK, KI, and MI. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Masayoshi Inoue.

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Competing interests

Kyoko Itoh has received grants for commissioned and joint research from SCREEN Holdings Co., Ltd., Kyoto, Japan. Hiroshi Ogi and Saya Shibata are employed by SCREEN Holdings Co., Ltd. The remaining authors declare no potential competing interests.

Ethics approval

This study was approved by the Institutional Review Board of the Kyoto Prefectural University of Medicine (ERB-C-931) and conducted in accordance with the Declaration of Helsinki.

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Written, informed consent was obtained from all patients involved in the present study.

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Furuya, T., Ishihara, S., Ogi, H. et al. Characteristic differences in the abundance of tumor-infiltrating lymphocytes and intratumoral developing T cells in thymoma, with special reference to PD-1 expression. Cancer Immunol Immunother 72, 2585–2596 (2023). https://doi.org/10.1007/s00262-023-03431-5

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