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Prognostic impact of programmed cell death 1 ligand 1 expression in human leukocyte antigen class I-positive hepatocellular carcinoma after curative hepatectomy

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
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Abstract

Background

Hepatocellular carcinoma (HCC) is one of the most common solid tumors worldwide. Surgery is potentially curative, but high recurrence rates worsen patient prognosis. The interaction between the proteins programmed cell death 1 (PD-1) and programmed cell death 1 ligand 1 (PD-L1) is an important immune checkpoint. The significance of PD-L1 expression and human leukocyte antigen class I (HLA class I), recognized by CD8 T cells, in the prognosis of patients with HCC remains to be determined.

Methods

We assessed the levels of PD-L1 and HLA class I expression on HCC samples from 80 patients who had undergone hepatectomy at our institution, and evaluated the correlations between PD-L1 and HLA class I expression and patient prognosis.

Results

High HLA class I expression was correlated with significantly better recurrence-free survival (RFS), but not overall survival (OS). Multivariate analysis showed that high HLA class I expression was an independent predictor of improved RFS. Low expression of PD-L1 on HCC tended to predict better OS, but the difference was not statistically significant. PD-L1 expression on HCC correlated with the number of CD163-positive macrophages and HLA class I expression with CD3-positive cell infiltration. Univariable and multivariable analyses showed that combined PD-L1 low/HLA class I high expression on HCCs was prognostic for improved OS and RFS.

Conclusions

PD-L1 status may be a good predictor of prognosis in HCC patients with high HLA class I expression. Novel therapies targeting the PD-L1/PD-1 pathway may improve the prognosis of patients with HCC.

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Abbreviations

HCC:

Hepatocellular carcinoma

PD-1:

Programmed cell death 1

PD-L1:

Programmed cell death 1 ligand 1

MHC:

Major histocompatibility complex

HLA:

Human leukocyte antigen

OS:

Overall survival

RFS:

Recurrence-free survival

pSTAT3:

Phosphorylated signal transducer and activator of transcription 3

TAM:

Tumor-associated macrophages

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Acknowledgments

This work was supported by a Grant-in-Aid from the Japan Society for the Promotion of Science (to Shirabe Ken; #24390320).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Yuichiroh Umemoto, Shinji Okano, Yoshihiro Matsumoto, Hidekazu Nakagawara, Rumi Matono, Shohei Yoshiya, Yo-ichi Yamashita, Tomoharu Yoshizumi, Toru Ikegami, Yuji Soejima, Ken Shirabe & Yoshihiko Maehara

  2. Department of Innovative Applied Oncology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Shinji Okano

  3. Division of Diagnostic Pathology, Kyushu University Hospital, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Shinji Okano

  4. Department of Immunology, Shimane University Faculty of Medicine, Izumo, Shimane, 693-8501, Japan

    Mamoru Harada

  5. Department of Anatomic Pathology, Pathological Sciences, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Shinichi Aishima & Yoshinao Oda

Authors
  1. Yuichiroh Umemoto
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  2. Shinji Okano
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  3. Yoshihiro Matsumoto
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  4. Hidekazu Nakagawara
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  5. Rumi Matono
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  6. Shohei Yoshiya
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  7. Yo-ichi Yamashita
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  8. Tomoharu Yoshizumi
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  9. Toru Ikegami
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  10. Yuji Soejima
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  11. Mamoru Harada
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  12. Shinichi Aishima
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  13. Yoshinao Oda
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  14. Ken Shirabe
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  15. Yoshihiko Maehara
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Corresponding author

Correspondence to Shinji Okano.

Additional information

Y. Umemoto and S. Okano contributed equally to this work.

Electronic supplementary material

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535_2014_933_MOESM1_ESM.pptx

Supplementary material 1 Fig. S1 (A, B) Sections were cut 4 μm thick, deparaffinized in xylene, and dehydrated in an ethanol series. For antigen retrieval, the specimens were pretreated in an autoclave in 0.01 M citrate buffer (pH 6.0). After blocking nonspecific binding of antibodies, the specimens were incubated with anti-HLA class 1 (EMR8-5, 1:500; MBL, Tokyo, Japan) followed by Alexa 647-conjugated anti-mouse IgG1 mAb, and with anti-CD3 (polyclonal rabbit antibody, Dako, Glostrup, Denmark) followed by Alexa 555-conjugated anti-rabbit antibody, each for 60 min at room temperature. The samples were subsequently, and counterstained with DAPI/anti-fade mounting medium (ProLong® Gold antifade reagent with DAPI, Life Technologies, Carlsbad, CA). Fluorescence was detected by confocal microscopy A1 (Nikon Instruments Inc., Tokyo, Japan). HLA class I was highly expressed on the hepatoma cell membranes of patient no. 1159, with this tumor showing prominent infiltration of CD3-positive T cells (A, original magnification × 200). HLA class I was hardly detected on the hepatoma cell membranes, but not on the endothelial cells, of patient no. 1164, with this tumor showing minimal infiltration of CD3-positive T cells (B, upper photograph, original magnification ×100). HLA class I was detected on the hepatoma cell membranes of another the site in patient L1164, with this area showing infiltration of CD3-positive T (B, lower photograph, original magnification ×100). (C) Tumor tissues were embedded in TissueTek cutting medium (Sakura Finetek, Tokyo, Japan) using Cryomold (Sakura Finetek), and 6-μm frozen tissue sections were fixed with cold acetone, washed with PBS, and incubated with 5 % skim milk in PBS for 1 h to blocking nonspecific binding. The sections were immunostained with phycoerythrin (PE)-conjugated anti-PD-L1 mAb (Biolegend, San Diego, CA), FITCconjugated HLA class I (BD Biosciences, San Diego, CA), and allophycocyanin (Apc)-conjugated CD163 mAb (Biolegend), and counterstained with DAPI/anti-fade mounting medium (ProLong® Gold antifade reagent with DAPI, Life Technologies). The sections were examined by microscopy (Biozero BZ-8000, KEYENCE Japan, Osaka, Japan), and composite images were created using the attached software. Hepatoma cells expressing PD-L1 (encircled area) were present at HCC sites infiltrated by CD163-positive tumor associated macrophages (arrow head). (PPTX 5910 kb)

535_2014_933_MOESM2_ESM.pptx

Supplementary material 2 Fig. S2 Sections showing HCC specimens derived from patients No.1875 (A, B, E, F; PD-L high/HLA class I high) and 1959 (C, D, G, H; PD-L1 low/HLA class I high) stained with H&E (A-D) and antibody to CD3 (E–H). Degenerative changes, necrosis, and necrosis of the HCC were prominent (arrow, C, D) and associated with massive infiltration of CD3-positive cells (G, H) in patient 1959, but were minimal (A, B) and associated with infiltration of a few CD3-positive cells only in the peripheral tissue of the tumor in patient 1875 (E, F). Despite similar characteristics of the tumor factors, patient 1959 showed no evidence of tumor recurrence, whereas patient 1959 showed HCC recurrence 180 days after curative hepatectomy. (PPTX 9249 kb)

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Umemoto, Y., Okano, S., Matsumoto, Y. et al. Prognostic impact of programmed cell death 1 ligand 1 expression in human leukocyte antigen class I-positive hepatocellular carcinoma after curative hepatectomy. J Gastroenterol 50, 65–75 (2015). https://doi.org/10.1007/s00535-014-0933-3

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  • DOI: https://doi.org/10.1007/s00535-014-0933-3

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