Abstract
Hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) is usually considered an inflammation-related cancer associated with chronic inflammation triggered by exposure to HBV and tumor antigens. T-cell exhaustion is implicated in immunosuppression of chronic infections and tumors. Although immunotherapies that enhance immune responses by targeting programmed cell death-1(PD-1)/PD-L1 are being applied to malignancies, these treatments have shown limited response rates, suggesting that additional inhibitory receptors are also involved in T-cell exhaustion and tumor outcome. Here, we analyzed peripheral blood samples and found that coexpression of PD-1 and T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT) was significantly upregulated on CD4+ and CD8+ T cells from patients with HBV-HCC compared with those from patients with chronic HBV or HBV-liver cirrhosis. Additionally, PD-1+ TIGIT+ CD8+ T-cell populations were elevated in patients with advanced stage and progressed HBV-HCC. Importantly, PD-1+ TIGIT+ CD8+ T-cell populations were negatively correlated with overall survival rate and progression-free survival rates. Moreover, we showed that PD-1+ TIGIT+ CD8+ T cells exhibit features of exhausted T cells, as manifested by excessive activation, high expression of other inhibitory receptors, high susceptibility to apoptosis, decreased capacity for cytokine secretion, and patterns of transcription factor expression consistent with exhaustion. In conclusion, PD-1+ TIGIT+ CD8+ T-cell populations are associated with accelerated disease progression and poor outcomes in HBV-HCC, which might not only have important clinical implications for prognosis but also provide a rationale for new targets in immunotherapy.





Similar content being viewed by others
Abbreviations
- AFP:
-
α-Fetoprotein
- ALB:
-
Albumin
- ALT:
-
Alanine aminotransferase
- AUROC:
-
Area under the receiver-operating characteristic
- BCLC:
-
Barcelona clinic liver cancer
- BTLA:
-
B- and T-lymphocyte attenuator
- CHB:
-
Chronic hepatitis B virus infection
- CTLA-4:
-
Cytotoxic T-lymphocyte antigen 4
- Eomes:
-
Eomesodermin
- FITC:
-
Fluorescein isothiocyanate
- HBeAg:
-
Hepatitis B e antigen
- HBV-HCC:
-
Hepatitis B virus-associated hepatocellular carcinoma
- HCV:
-
Hepatitis C virus
- HIV:
-
Human immunodeficiency virus
- HR:
-
Hazard ratio
- IFN-γ:
-
Interferon gamma
- LAG-3:
-
Lymphocyte-activation gene 3
- LC:
-
Liver cirrhosis
- MELD:
-
Model for end-stage liver disease
- NLR:
-
Neutrophil–lymphocyte ratio
- NK:
-
Natural killer cells
- OS:
-
Overall survival
- PBMC:
-
Peripheral blood mononuclear cell
- PD-1:
-
Programmed cell death-1
- PD-L1:
-
Programmed death-ligand 1
- PFS:
-
Progression-free survival
- TBIL:
-
Total bilirubin
- TCM:
-
Central memory T cells
- TEM:
-
Effector memory T cells
- TEMRA:
-
Terminally differentiated effector T cells
- TIGIT:
-
T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain
- TILs:
-
Tumor-infiltrating lymphocytes
- TIM-3:
-
T-cell immunoglobulin domain and mucin domain 3
- TNF-α:
-
Tumor necrosis factor
References
Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA 65(2):87–108
Parkin DM (2006) The global health burden of infection-associated cancers in the year 2002. Int J Cancer 118(12):3030–3044
de Martel C, Maucort-Boulch D, Plummer M, Franceschi S (2015) World-wide relative contribution of hepatitis B and C viruses in hepatocellular carcinoma. Hepatology 62(4):1190–1200
Papatheodoridis GV, Chan HL, Hansen BE, Janssen HL, Lampertico P (2015) Risk of hepatocellular carcinoma in chronic hepatitis B: assessment and modification with current antiviral therapy. J Hepatol 62(4):956–967
Wang FS, Fan JG, Zhang Z, Gao B, Wang HY (2014) The global burden of liver disease: the major impact of China. Hepatology 60(6):2099–2108
De Palma M, Biziato D, Petrova TV (2017) Microenvironmental regulation of tumour angiogenesis. Nat Rev Cancer 17(8):457–474
Kitamura T, Qian BZ, Pollard JW (2015) Immune cell promotion of metastasis. Nat Rev Immunol 15(2):73–86
Munn DH, Bronte V (2016) Immune suppressive mechanisms in the tumor microenvironment. Curr Opin Immunol 39:1–6
Wherry EJ (2011) T cell exhaustion. Nat Immunol 12(6):492–499
Pauken KE, Wherry EJ (2015) Overcoming T cell exhaustion in infection and cancer. Trends Immunol 36(4):265–276
Wherry EJ, Kurachi M (2015) Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol 15(8):486–499
Reck M, Rodriguez-Abreu D, Robinson AG, Hui R, Csoszi T, Fulop A et al (2016) Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer. N Engl J Med 375(19):1823–1833
Robert C, Ribas A, Wolchok JD, Hodi FS, Hamid O, Kefford R et al (2014) Anti-programmed-death-receptor-1 treatment with pembrolizumab in ipilimumab-refractory advanced melanoma: a randomised dose-comparison cohort of a phase 1 trial. Lancet 384(9948):1109–1117
Weber JS, D’Angelo SP, Minor D, Hodi FS, Gutzmer R, Neyns B et al (2015) Nivolumab versus chemotherapy in patients with advanced melanoma who progressed after anti-CTLA-4 treatment (CheckMate 037): a randomised, controlled, open-label, phase 3 trial. Lancet Oncol 16(4):375–384
Motzer RJ, Escudier B, McDermott DF, George S, Hammers HJ, Srinivas S et al (2015) Nivolumab versus everolimus in advanced renal-cell carcinoma. N Engl J Med 373(19):1803–1813
El-Khoueiry AB, Sangro B, Yau T et al (2017) Nivolumab in patients with advanced hepatocellular carcinoma (CheckMate 040): an open-label, non-comparative, phase 1/2 dose escalation and expansion trial[J]. The Lancet 389(10088):2492–2502
Inozume T, Yaguchi T, Furuta J, Harada K, Kawakami Y, Shimada S (2016) Melanoma cells control antimelanoma CTL responses via interaction between TIGIT and CD155 in the effector phase. J Invest Dermatol 136(1):255–263
Joller N, Lozano E, Burkett PR, Patel B, Xiao S, Zhu C et al (2014) Treg cells expressing the coinhibitory molecule TIGIT selectively inhibit proinflammatory Th1 and Th17 cell responses. Immunity 40(4):569–581
Kurtulus S, Sakuishi K, Ngiow SF, Joller N, Tan DJ, Teng MW et al (2015) TIGIT predominantly regulates the immune response via regulatory T cells. J Clin Investig 125(11):4053–4062
Chan CJ, Martinet L, Gilfillan S, Souza-Fonseca-Guimaraes F, Chow MT, Town L et al (2014) The receptors CD96 and CD226 oppose each other in the regulation of natural killer cell functions. Nat Immunol 15(5):431–438
Chauvin JM, Pagliano O, Fourcade J, Sun Z, Wang H, Sander C et al (2015) TIGIT and PD-1 impair tumor antigen-specific CD8(+) T cells in melanoma patients. J Clin Investig 125(5):2046–2058
Johnston RJ, Comps-Agrar L, Hackney J, Yu X, Huseni M, Yang Y et al (2014) The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer Cell 26(6):923–937
Liu X, Li M, Wang X et al (2019) Effects of adjuvant traditional Chinese medicine therapy on long-term survival in patients with hepatocellular carcinoma. Phytomedicine 62:152930
Song Y, Wang B, Song R et al (2018) T-cell immunoglobulin and ITIM domain contributes to CD 8 + T-cell immunosenescence. Aging Cell 17(2):e12716
Motomura T, Shirabe K, Mano Y, Muto J, Toshima T, Umemoto Y et al (2013) Neutrophil-lymphocyte ratio reflects hepatocellular carcinoma recurrence after liver transplantation via inflammatory microenvironment. J Hepatol 58(1):58–64
Mano Y, Shirabe K, Yamashita Y, Harimoto N, Tsujita E, Takeishi K et al (2013) Preoperative neutrophil-to-lymphocyte ratio is a predictor of survival after hepatectomy for hepatocellular carcinoma: a retrospective analysis. Ann Surg 258(2):301–305
Templeton AJ, McNamara MG, Seruga B, Vera-Badillo FE, Aneja P, Ocana A et al (2014) Prognostic role of neutrophil-to-lymphocyte ratio in solid tumors: a systematic review and meta-analysis. J Natl Cancer Inst 106(6):24
Cho H, Hur HW, Kim SW, Kim SH, Kim JH, Kim YT et al (2009) Pre-treatment neutrophil to lymphocyte ratio is elevated in epithelial ovarian cancer and predicts survival after treatment. Cancer Immunol Immunother CII 58(1):15–23
Jankovic V, Messaoudi I, Nikolich-Zugich J (2003) Phenotypic and functional T-cell aging in rhesus macaques (Macaca mulatta): differential behavior of CD4 and CD8 subsets. Blood 102(9):3244–3251
Blackburn SD, Shin H, Haining WN, Zou T, Workman CJ, Polley A et al (2009) Coregulation of CD8 + T cell exhaustion by multiple inhibitory receptors during chronic viral infection. Nat Immunol 10(1):29–37
Barber DL, Wherry EJ, Masopust D, Zhu B, Allison JP, Sharpe AH et al (2006) Restoring function in exhausted CD8 T cells during chronic viral infection. Nature 439(7077):682–687
Buggert M, Tauriainen J, Yamamoto T, Frederiksen J, Ivarsson MA, Michaelsson J et al (2014) T-bet and Eomes are differentially linked to the exhausted phenotype of CD8 + T cells in HIV infection. PLoS Pathog 10(7):e1004251
Doering TA, Crawford A, Angelosanto JM, Paley MA, Ziegler CG, Wherry EJ (2012) Network analysis reveals centrally connected genes and pathways involved in CD8 + T cell exhaustion versus memory. Immunity 37(6):1130–1144
Shi F, Shi M, Zeng Z, Qi RZ, Liu ZW, Zhang JY et al (2011) PD-1 and PD-L1 upregulation promotes CD8(+) T-cell apoptosis and postoperative recurrence in hepatocellular carcinoma patients. Int J Cancer 128(4):887–896
Zeng Z, Shi F, Zhou L, Zhang MN, Chen Y, Chang XJ et al (2011) Upregulation of circulating PD-L1/PD-1 is associated with poor post-cryoablation prognosis in patients with HBV-related hepatocellular carcinoma. PLoS One 6(9):e23621
Gao Q, Wang XY, Qiu SJ, Yamato I, Sho M, Nakajima Y et al (2009) Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma. Clin Cancer Res 15(3):971–979
Zhao Q, Huang ZL, He M, Gao Z, Kuang DM (2016) BTLA identifies dysfunctional PD-1-expressing CD4(+) T cells in human hepatocellular carcinoma. Oncoimmunology 5(12):e1254855
Stanietsky N, Rovis TL, Glasner A, Seidel E, Tsukerman P, Yamin R et al (2013) Mouse TIGIT inhibits NK-cell cytotoxicity upon interaction with PVR. Eur J Immunol 43(8):2138–2150
Liu S, Zhang H, Li M, Hu D, Li C, Ge B et al (2013) Recruitment of Grb2 and SHIP1 by the ITT-like motif of TIGIT suppresses granule polarization and cytotoxicity of NK cells. Cell Death Differ 20(3):456–464
Li M, Xia P, Du Y, Liu S, Huang G, Chen J et al (2014) T-cell immunoglobulin and ITIM domain (TIGIT) receptor/poliovirus receptor (PVR) ligand engagement suppresses interferon-gamma production of natural killer cells via beta-arrestin 2-mediated negative signaling. J Biol Chem 289(25):17647–17657
Yu X, Harden K, Gonzalez LC, Francesco M, Chiang E, Irving B et al (2009) The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat Immunol 10(1):48–57
Liu X, He L, Han J, Wang L, Li M, Jiang Y et al (2017) Association of neutrophil-lymphocyte ratio and T lymphocytes with the pathogenesis and progression of HBV-associated primary liver cancer. PLoS One 12(2):e0170605
Wherry EJ, Blattman JN, Murali-Krishna K, van der Most R, Ahmed R (2003) Viral persistence alters CD8 T-cell immunodominance and tissue distribution and results in distinct stages of functional impairment. J Virol 77(8):4911–4927
Huang AC, Postow MA, Orlowski RJ, Mick R, Bengsch B, Manne S et al (2017) T-cell invigoration to tumour burden ratio associated with anti-PD-1 response. Nature 545(7652):60–65
Ahn E, Youngblood B, Lee J, Lee J, Sarkar S, Ahmed R (2016) Demethylation of the PD-1 promoter is imprinted during the effector phase of CD8 T cell exhaustion. J Virol 90(19):8934–8946
Inozume T, Yaguchi T, Furuta J, Harada K, Kawakami Y, Shimada S (2016) Melanoma cells control antimelanoma CTL responses via interaction between TIGIT and CD155 in the effector phase. J Invest Dermatol 136(1):255–263
Chew V, Lai L, Pan L, Lim CJ, Li J, Ong R et al (2017) Delineation of an immunosuppressive gradient in hepatocellular carcinoma using high-dimensional proteomic and transcriptomic analyses. Proc Natl Acad Sci USA 114(29):E5900–E5909
Acknowledgements
We would like to thank Lihua Yu and Dongdong Zhou for their assistance with the follow-up of survival information in HBV-HCC patients.
Funding
This work was supported by Application of Clinical Features of Capital City of Science and Technology Commission (No. Z171100001017082); the Fund for Beijing Science & Technology Development of TCM (No. JJ2016-14); the Fund of Special research of TCM in Capital City (17ZY02), the National Key Sci-Tech Special Project of China (No. 2018ZX10302207).
Author information
Authors and Affiliations
Contributions
ZY and YK designed the study; XL and YK performed experiments and wrote the manuscript; ML, XW, and ZD provided patient material and performed experiments; YJ and XW were responsible for the interpretation of data and revision of the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interests.
Ethics approval and consent standards
The study was approved by the ethics committee of Beijing Ditan Hospital, Capital Medical University (2016-1-28).
Informed consent
Each patient and healthy donors signed an informed consent. They all agreed to use their specimens and clinical information for this research.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Liu, X., Li, M., Wang, X. et al. PD-1+ TIGIT+ CD8+ T cells are associated with pathogenesis and progression of patients with hepatitis B virus-related hepatocellular carcinoma. Cancer Immunol Immunother 68, 2041–2054 (2019). https://doi.org/10.1007/s00262-019-02426-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00262-019-02426-5