Advertisement

Flow Cytometry Analysis of Surface PD-L1 Expression Induced by IFNγ and Romidepsin in Ovarian Cancer Cells

  • Sveta Padmanabhan
  • Yue Zou
  • Ivana VancurovaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2108)

Abstract

Expression of programmed death ligand-1 (PD-L1, CD274) on cancer cells is regulated by interferon-γ (IFNγ) signaling as well as by epigenetic mechanisms. By binding to PD-1 on cytotoxic T cells, PD-L1 inhibits T cell-mediated antitumor responses, resulting in immune escape. This chapter describes analysis of the surface PD-L1 expression in ovarian cancer (OC) cells using flow cytometry (FC). Our data demonstrate that the surface PD-L1 expression in OC cells is induced by IFNγ as well as by the class I histone deacetylase (HDAC) inhibition by romidepsin, suggesting that class I HDAC inhibition might provide a useful strategy to modulate the PD-L1 levels on OC cells.

Key words

HDAC HDAC inhibition Interferon-γ Immune escape Flow cytometry Ovarian cancer PD-L1 Romidepsin 

Notes

Acknowledgment

This work was supported by National Institutes of Health Grant CA202775 (to I.V.).

References

  1. 1.
    Freeman GJ, Long AJ, Iwai Y et al (2000) Engagement of the PD-1 immuno-inhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med 192:1027–1034CrossRefGoogle Scholar
  2. 2.
    Iwai Y, Ishida M, Tanaka Y et al (2002) Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade. Proc Natl Acad Sci U S A 99:12293–12297CrossRefGoogle Scholar
  3. 3.
    Loke P, Allison JP (2003) PD-L1 and PD-L2 are differentially regulated by Th1 and Th2 cells. Proc Natl Acad Sci U S A 100:5336–5341CrossRefGoogle Scholar
  4. 4.
    Azuma T, Yao S, Zhu G et al (2008) B7-H1 is a ubiquitous antiapoptotic receptor on cancer cells. Blood 111:3635–3643CrossRefGoogle Scholar
  5. 5.
    Chang CH, Qiu J, O’Sullivan D et al (2015) Metabolic competition in the tumor microenvironment is a driver of cancer progression. Cell 162:1229–1241CrossRefGoogle Scholar
  6. 6.
    Clark CA, Gupta HB, Sareddy G et al (2016) Tumor-intrinsic PD-L1 signals regulate cell growth, pathogenesis, and autophagy in ovarian cancer and melanoma. Cancer Res 76:6964–6974CrossRefGoogle Scholar
  7. 7.
    Clark CA, Gupta HB, Curiel TJ (2017) Tumor cell-intrinsic CD274/PD-L1: a novel metabolic balancing act with clinical potential. Autophagy 13:987–988CrossRefGoogle Scholar
  8. 8.
    Garcia-Diaz A, Shin DS, Moreno BH et al (2017) Interferon receptor signaling pathways regulating PD-L1 and PD-L2 expression. Cell Rep 19:1189–1201CrossRefGoogle Scholar
  9. 9.
    Ivashkiv LB (2018) IFNγ: signalling, epigenetics and roles in immunity, metabolism, disease and cancer immunotherapy. Nat Rev Immunol 18:545–558CrossRefGoogle Scholar
  10. 10.
    Woods DM, Sodré AL, Villagra A et al (2015) HDAC inhibition upregulates PD-1 ligands in melanoma and augments immunotherapy with PD-1 blockade. Cancer Immunol Res 3:1375–1385CrossRefGoogle Scholar
  11. 11.
    Cacan E (2017) Epigenetic-mediated immune suppression of positive co-stimulatory molecules in chemoresistant ovarian cancer cells. Cell Biol Int 41:328–339CrossRefGoogle Scholar
  12. 12.
    Briere D, Sudhakar N, Woods DM et al (2018) The class I/IV HDAC inhibitor mocetinostat increases tumor antigen presentation, decreases immune suppressive cell types and augments checkpoint inhibitor therapy. Cancer Immunol Immunother 67:381–392CrossRefGoogle Scholar
  13. 13.
    Iwasa M, Harada T, Oda A et al (2019) PD-L1 upregulation in myeloma cells by panobinostat in combination with interferon-γ. Oncotarget 10:1903–1917CrossRefGoogle Scholar
  14. 14.
    Llopiz D, Ruiz M, Villanueva L et al (2019) Enhanced anti-tumor efficacy of checkpoint inhibitors in combination with the histone deacetylase inhibitor Belinostat in a murine hepatocellular carcinoma model. Cancer Immunol Immunother 68:379–393CrossRefGoogle Scholar
  15. 15.
    Terranova-Barberio M, Thomas S, Ali N et al (2017) HDAC inhibition potentiates immunotherapy in triple negative breast cancer. Oncotarget 8:114156–114172PubMedPubMedCentralGoogle Scholar
  16. 16.
    Bae J, Hideshima T, Tai YT et al (2018) Histone deacetylase (HDAC) inhibitor ACY241 enhances anti-tumor activities of antigen-specific central memory cytotoxic T lymphocytes against multiple myeloma and solid tumors. Leukemia 32:1932–1947CrossRefGoogle Scholar
  17. 17.
    Knox T, Sahakian E, Banik D et al (2019) Selective HDAC6 inhibitors improve anti-PD-1 immune checkpoint blockade therapy by decreasing the anti-inflammatory phenotype of macrophages and down-regulation of immunosuppressive proteins in tumor cells. Sci Rep 9(1):6136CrossRefGoogle Scholar
  18. 18.
    Armbruster S, Coleman RL, Rauh-Hain JA (2018) Management and treatment of recurrent epithelial ovarian cancer. Hematol Oncol Clin North Am 32:965–982CrossRefGoogle Scholar
  19. 19.
    Chodon T, Lugade AA, Battaglia S, Odunsi K (2018) Emerging role and future directions of immunotherapy in advanced ovarian cancer. Hematol Oncol Clin North Am 32:1025–1039CrossRefGoogle Scholar
  20. 20.
    Vaughan S, Coward JI, Bast RC Jr et al (2011) Rethinking ovarian cancer: recommendations for improving outcomes. Nat Rev Cancer 11:719–725CrossRefGoogle Scholar
  21. 21.
    Abiko K, Matsumura N, Hamanishi J et al (2015) IFN-γ from lymphocytes induces PD-L1 expression and promotes progression of ovarian cancer. Br J Cancer 112:1501–1509CrossRefGoogle Scholar
  22. 22.
    Mandai M, Hamanishi J, Abiko K et al (2016) Dual faces of IFNγ in cancer progression: a role of PD-L1 induction in the determination of pro- and antitumor immunity. Clin Cancer Res 22:2329–2334CrossRefGoogle Scholar
  23. 23.
    Zou Y, Uddin MM, Padmanabhan S et al (2018) The proto-oncogene Bcl3 induces immune checkpoint PD-L1 expression, mediating proliferation of ovarian cancer cells. J Biol Chem 293:15483–15496CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Biological SciencesSt. John’s UniversityQueensUSA

Personalised recommendations