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Multispectral quantitative immunohistochemical analysis of tumor-infiltrating lymphocytes in relation to programmed death-ligand 1 expression in triple-negative breast cancer

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Abstract

Background

Programmed death-ligand 1 (PD-L1) expression on immune cells (ICs) is a predictive marker for PD-L1 checkpoint blockade in patients with triple-negative breast cancer (TNBC). However, the level of PD-L1 expression and the percentage of cells that are PD-L1+ are continuous variables not dichotomous variables for tumor-infiltrating lymphocytes (TILs) and other cells.

Methods

Multiplexed immunohistochemistry was applied to 31 archived surgical specimens from untreated TNBC patients. TIL levels were visually scored, and CD8+ T cells and PD-L1+ ICs were quantified using an automated multispectral imaging system. PD-L1 expression was assessed within a multiplexed context (CD8 combined spectral composite).

Results

The mean value of stromal TILs (i.e., the percentage of the stromal area with a dese mononuclear infiltrate) was 20%. The frequency of patients with PD-L1-positive tumor cells (TC) and ICs was 38.7% and 32.2%, respectively, with a significant association between them. TIL levels were correlated with CD8+ T cell infiltration in the stroma (Spearman r = 0.795, p < 0.0001). PD-L1 expression on IC was significantly associated with TIL levels (Spearman r = 0.790, p < 0.001) and infiltration of CD8+ T cells (Spearman r = 0.683, p < 0.0001).

Conclusions

The level of PD-L1 on IC was correlated with the level of PD-L1 on TC as well as TIL levels and infiltration of CD8+ T cells. These results suggest that high PD-L1 on IC may reflect T cell-inflamed tumors with the amount of TILs present, including the CD8+ T cells required for anti-tumor responses.

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Acknowledgements

We thank Mamoru Nomura at Perkin Elmer Japan for the excellent technical assistance.

Funding

This study was supported by a grant from the Japanese Breast Cancer Society.

Author information

Authors and Affiliations

  1. Breast Surgery, Kansai Medical University Hospital, Hirakata, Japan

    Tomoharu Sugie

  2. Department of Pathology (Medical Research Center), Institute of Medical Science, Tokyo Medical University, Tokyo, Japan

    Eiichi Sato

  3. Department of Breast and Endocrine Surgical Oncology, Tohoku University Graduate School of Medicine, Sendai, Japan

    Minoru Miyashita

  4. Department of Pathology and Clinical Medicine, Kurume University Medical Center, Kurume, Japan

    Rin Yamaguchi

  5. Department of Diagnostic Pathology, Nippon Medical School Hospital, Tokyo, Japan

    Takashi Sakatani

  6. Department of Pathology, Mie University Hospital, Tsu, Japan

    Yuji Kozuka

  7. Department of Diagnostic Pathology, Shiga University of Medical Science, Otsu, Japan

    Suzuko Moritani

  8. Breast Surgery, Graduate School of Medicine Kyoto University, Kyoto, Japan

    Eiji Suzuki

  9. Cancer Immunology Data Multi-level Integration Unit, Medical Science Innovation, Hub Program, RIKEN, Tokyo, Japan

    Kazuhiro Kakimi

  10. Department of Diagnostic Pathology, Kumamoto University Hospital, Kumamoto, Japan

    Yoshiki Mikami

  11. Department of Pathology, Kawasaki Medical School, Kurashiki, Japan

    Takuya Moriya

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  1. Tomoharu Sugie
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Correspondence to Tomoharu Sugie.

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All procedures performed in this study were in accordance with the ethical standards of the Kawasaki Medical School Ethical Committee.

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Sugie, T., Sato, E., Miyashita, M. et al. Multispectral quantitative immunohistochemical analysis of tumor-infiltrating lymphocytes in relation to programmed death-ligand 1 expression in triple-negative breast cancer. Breast Cancer 27, 519–526 (2020). https://doi.org/10.1007/s12282-020-01110-2

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