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Inhibition of T-cell responses by intratumoral hepatic stellate cells contribute to migration and invasion of hepatocellular carcinoma

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Abstract

The stroma of hepatocellular carcinoma (HCC) is markedly infiltrated with activated hepatic stellate cells (HSCs), and associated invasion and metastasis of HCC. However, little is known of the role of HSCs in immune responses in HCC. The Buffalo rat HCC model was established. Quiescent HSCs (qHSCs) and intratumoral HSCs (tHSCs) were isolated. Surface molecules of tHSC were detected by flow cytometry, and gene expression was analyzed by fluorescence quantitative RT-PCR. T cell proliferation was monitored by [3H]-thymidine (3H-TdR) incorporation into DNA, and cytotoxic activity was assessed by measuring the release of 51Cr. The level of cytokine expression by T cells was measured by enzyme-linked immunosorbent assay. T cell apoptosis was detected by double-stained terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and anti-CD3 antibodies. The migration and invasion of HCC was observed by transwell experiments. tHSCs express low levels of major histocompatibility complex (MHC) class I, MHC class II, and costimulatory molecules, and produce varying levels of cytokines. Addition of the tHSCs suppressed thymidine uptake by T cells that were stimulated by alloantigens or by anti-CD3-mediated T-cell receptor ligation. The tHSC-induced T-cell hyporesponsiveness was associated with enhanced T-cell apoptosis, and contributed to the migration and invasion of hepatoma cell. tHSCs was associated with markedly enhanced expression of B7-H1. Blockade of B7-H1/PD-1 ligation significantly reduced HSC immunomodulatory activity, and hepatoma cell migration and invasion. tHSCs can induce T cell apoptosis, suggesting an important role for B7-H1. The interactions between tHSCs and T cells may contribute to hepatic immune tolerance and invasion and migration of HCC.

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Abbreviations

HSCs:

Hepatic stellate cells

tHSCs:

Intratumoral hepatic stellate cells

ECM:

Extracellular matrix

α-SMA:

α-Smooth muscle actin

qHSCs:

Quiescent HSCs

HCC:

Hepatocellular carcinoma

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling

[3H]-TdR:

[3H]-Thymidine

IFN:

Interferon

DC:

Dendritic cell

CTLs:

Specific cytotoxic T lymphocytes

APCs:

Antigen presenting cells

GM-CSF:

Granulocyte-macrophage colony-stimulating factor

IL:

Interleukin

References

  1. Gressner AM, Weiskirchen R, Breitkopf K et al (2002) Roles of TGF-beta in hepatic fibrosis. Front Biosci 7:d793–d807

    Article  PubMed  CAS  Google Scholar 

  2. Svegliati-Baroni G, De Minicis S, Marzioni M (2008) Hepatic fibrogenesis in response to chronic liver injury: novel insights on the role of cell-to-cell interaction and transition. Liver Int 28:1052–1064

    Article  PubMed  CAS  Google Scholar 

  3. Marra F, DeFranco R, Grappone C et al (1999) Expression of monocyte chemotactic protein-1 precedes monocyte recruitment in a rat model of acute liver injury, and is modulated by vitamin E. J Investig Med 47:66–75

    PubMed  CAS  Google Scholar 

  4. Lu L, Qian S, Hershberger PA et al (1997) Fas ligand (CD95L) and B7 expression on dendritic cells provide counter-regulatory signals for T cell survival and proliferation. J Immunol 158:5676–5684

    PubMed  CAS  Google Scholar 

  5. Lu L, Woo J, Rao AS et al (1994) Propagation of dendritic cell progenitors from normal mouse liver using granulocyte/macrophage colony-stimulating factor and their maturational development in the presence of type-1 collagen. J Exp Med 179:1823–1834

    Article  PubMed  CAS  Google Scholar 

  6. Friedman SL (1993) Seminars in medicine of the Beth Israel Hospital, Boston. The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies. N Engl J Med 328:1828–1835

    Article  PubMed  CAS  Google Scholar 

  7. Kobayashi S, Seki S, Kawada N et al (2003) Apoptosis of T cells in the hepatic fibrotic tissue of the rat: a possible inducing role of hepatic myofibroblast-like cells. Cell Tissue Res 311:353–364

    PubMed  Google Scholar 

  8. Yu MC, Chen CH, Liang X et al (2004) Inhibition of T-cell responses by hepatic stellate cells via B7-H1-mediated T-cell apoptosis in mice. Hepatology 40:1312–1321

    Article  PubMed  CAS  Google Scholar 

  9. Chen CH, Kuo LM, Chang Y et al (2006) In vivo immune modulatory activity of hepatic stellate cells in mice. Hepatology 44:1171–1181

    Article  PubMed  CAS  Google Scholar 

  10. Cabrera R, Ararat M, Cao M et al (2010) Hepatocellular carcinoma immunopathogenesis: clinical evidence for global T cell defects and an immunomodulatory role for soluble CD25 (sCD25). Dig Dis Sci 55:484–495

    Article  PubMed  CAS  Google Scholar 

  11. Kuang DM, Zhao Q, Peng C et al (2009) Activated monocytes in peritumoral stroma of hepatocellular carcinoma foster immune privilege and disease progression through PD-L1. J Exp Med 206:1327–1337

    Article  PubMed  CAS  Google Scholar 

  12. Terada T, Makimoto K, Terayama N et al (1996) Alpha-smooth muscle actin-positive stromal cells in cholangiocarcinomas, hepatocellular carcinomas and metastatic liver carcinomas. J Hepatol 24:706–712

    Article  PubMed  CAS  Google Scholar 

  13. Aishima S, Nishihara Y, Iguchi T et al (2008) Lymphatic spread is related to VEGF-C expression and D2-40-positive myofibroblasts in intrahepatic cholangiocarcinoma. Mod Pathol 21:256–264

    Article  PubMed  CAS  Google Scholar 

  14. Desmoulière A, Guyot C, Gabbiani G (2004) The stroma reaction myofibroblast: a key player in the control of tumor cell behavior. Int J Dev Biol 48:509–517

    Article  PubMed  Google Scholar 

  15. Yang R, Rescorla FJ, Reilly CR et al (1992) A reproducible rat liver cancer model for experimental therapy: introducing a technique of intrahepatic tumor implantation. J Surg Res 52:193–198

    Article  PubMed  CAS  Google Scholar 

  16. Kristensen DB, Kawada N, Imamura K et al (2000) Proteome analysis of rat hepatic stellate cells. Hepatology 32:268–277

    Article  PubMed  CAS  Google Scholar 

  17. Liu C, Gaca MD, Swenson ES et al (2003) Smads 2 and 3 are differentially activated by transforming growth factor-beta (TGF-beta) in quiescent and activated hepatic stellate cells: constitutive nuclear localization of Smads in activated cells is TGF-beta-independent. J Biol Chem 278:11721–11728

    Article  PubMed  CAS  Google Scholar 

  18. Lu L, McCaslin D, Starzl TE et al (1995) Bone marrow-derived dendritic cell progenitors (NLDC 145+, MHC class II+, B7-1dim, B7-2-) induce alloantigen-specific hyporesponsiveness in murine T lymphocytes. Transplantation 60:1539–1545

    Article  PubMed  CAS  Google Scholar 

  19. Matzinger P (1991) The JAM test: a simple assay for DNA fragmentation and cell death. J Immunol Methods 145:185–192

    Article  PubMed  CAS  Google Scholar 

  20. Hatfield P, Merrick AE, West E et al (2008) Optimization of dendritic cell loading with tumor cell lysates for cancer immunotherapy. J Immunother 31:620–632

    Article  PubMed  Google Scholar 

  21. Curiel TJ, Wei S, Dong H et al (2003) Blockade of B7-H1 improves myeloid dendritic cell-mediated antitumor immunity. Nat Med 9:562–567

    Article  PubMed  CAS  Google Scholar 

  22. Maher JJ (2001) Interactions between hepatic stellate cells and the immune system. Semin Liver Dis 21:417–426

    Article  PubMed  CAS  Google Scholar 

  23. Uemura T, Gandhi CR (2001) Inhibition of DNA synthesis in cultured hepatocytes by endotoxin-conditioned medium of activated stellate cells is transforming growth factor-beta and nitric oxide-independent. Br J Pharmacol 133:1125–1133

    Article  PubMed  CAS  Google Scholar 

  24. Kalinski P (2001) Dendritic cell-related immunoregulation: signals and mediators. In: Lotze MT, Thomson AW (eds) Dendritic cells, 2nd edn. Academic Press, London, pp 62–66

    Google Scholar 

  25. Wang SC, Ohata M, Schrum L et al (1998) Expression of interleukin-10 by in vitro and in vivo activated hepatic stellate cells. J Biol Chem 273:302–308

    Article  PubMed  CAS  Google Scholar 

  26. Thompson KC, Trowern A, Fowell A et al (1998) Primary rat and mouse hepatic stellate cells express the macrophage inhibitor cytokine interleukin-10 during the course of activation in vitro. Hepatology 28:1518–1524

    Article  PubMed  CAS  Google Scholar 

  27. Savage CO, Hughes CC, Pepinsky RB et al (1991) Endothelial cell lymphocyte function-associated antigen-3 and an unidentified ligand act in concert to provide costimulation to human peripheral blood CD4+ T cells. Cell Immunol 137:150–163

    Article  PubMed  CAS  Google Scholar 

  28. Dong H, Zhu G, Tamada K, Chen L (1999) B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion. Nat Med 5:1365–1369

    Article  PubMed  CAS  Google Scholar 

  29. Freeman GJ, Long AJ, Iwai Y et al (2000) Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med 192:1027–1034

    Article  PubMed  CAS  Google Scholar 

  30. Carter L, Fouser LA, Jussif J et al (2002) PD-1:PD-L inhibitory pathway affects both CD4(+) and CD8(+) T cells and is overcome by IL-2. Eur J Immunol 32:634–643

    Article  PubMed  CAS  Google Scholar 

  31. Agata Y, Kawasaki A, Nishimura H et al (1996) Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int Immunol 8:765–772

    Article  PubMed  CAS  Google Scholar 

  32. Larmonier N, Cathelin D, Larmonier C et al (2007) The inhibition of TNF-alpha anti-tumoral properties by blocking antibodies promotes tumor growth in a rat model. Exp Cell Res 313:2345–2355

    Article  PubMed  CAS  Google Scholar 

  33. Viñas O, Bataller R, Sancho-Bru P et al (2003) Human hepatic stellate cells show features of antigen-presenting cells and stimulate lymphocyte proliferation. Hepatology 38:919–929

    PubMed  Google Scholar 

  34. Winau F, Hegasy G, Weiskirchen R et al (2007) Ito cells are liver-resident antigen-presenting cells for activating T cell responses. Immunity 26:117–129

    Article  PubMed  CAS  Google Scholar 

  35. Faouzi S, Lepreux S, Bedin C et al (1999) Activation of cultured rat hepatic stellate cells by tumoral hepatocytes. Lab Invest 79:485–493

    PubMed  CAS  Google Scholar 

  36. Kobayashi S, Seki S, Kawada N et al (2003) Apoptosis of T cells in the hepatic fibrotic tissue of the rat: a possible inducing role of hepatic myofibroblast-like cells. Cell Tissue Res 311:353–364

    PubMed  Google Scholar 

  37. Dong H, Strome SE, Salomao DR et al (2002) Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med 8:793–800

    PubMed  CAS  Google Scholar 

  38. Dong H, Gefeng Z, Tamada K et al (2004) B7-H1 determines accumulation and deletion of intrahepatic CD8+ T lymphocytes. Immunity 20:327–336

    Article  PubMed  CAS  Google Scholar 

  39. Kanai T, Totsuka T, Uraushihara K et al (2003) Blockade of B7-H1 suppresses the development of chronic intestinal inflammation. J Immunol 171:4156–4163

    PubMed  CAS  Google Scholar 

  40. Binker MG, Binker-Cosen AA, Gaisano HY et al (2011) TGF-β1 increases invasiveness of SW1990 cells through Rac1/ROS/NF-κB/IL-6/MMP-2. Biochem Biophys Res Commun 405:140–145

    Article  PubMed  CAS  Google Scholar 

  41. Kim BC, Kim HT, Mamura M et al (2002) Tumor necrosis factor induces apoptosis in hepatoma cells by increasing Ca(2+) release from the endoplasmic reticulum and suppressing Bcl-2 expression. J Biol Chem 277:31381–31389

    Article  PubMed  CAS  Google Scholar 

  42. Kinter AL, Godbout EJ, McNally JP, Sereti I et al (2008) The common gamma-chain cytokines IL-2, IL-7, IL-15, and IL-21 induce the expression of programmed death-1 and its ligands. J Immunol 181:6738–6746

    PubMed  CAS  Google Scholar 

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Acknowledgment

This work was supported by the National Key Basic Research Project, No.2004CB518708A, China.

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Authors and Affiliations

  1. Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Rongxin Chen, Sheng-Long Ye, Ruixia Sun, Jun Chen & Yan Zhao

  2. Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai, 200032, China

    Rongxin Chen, Sheng-Long Ye, Ruixia Sun, Jun Chen & Yan Zhao

  3. Department of Oncology, Huaian Second People’s Hospital, Xuzhou Medical College, Huaian, 223002, China

    Yunhong Xia

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  1. Yunhong Xia
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Correspondence to Sheng-Long Ye.

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Xia, Y., Chen, R., Ye, SL. et al. Inhibition of T-cell responses by intratumoral hepatic stellate cells contribute to migration and invasion of hepatocellular carcinoma. Clin Exp Metastasis 28, 661–674 (2011). https://doi.org/10.1007/s10585-011-9399-3

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  • DOI: https://doi.org/10.1007/s10585-011-9399-3

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