Assessment of Antitumor T-Cell Responses by Flow Cytometry After Coculture of Tumor Cells with Autologous Tumor-Infiltrating Lymphocytes

  • Jeroen MeliefEmail author
  • Stina Wickström
  • Rolf Kiessling
  • Yago Pico de Coaña
Part of the Methods in Molecular Biology book series (MIMB, volume 1913)


As a consequence of the ever-increasing use of immunotherapy for the treatment of cancer, interest in the direct interaction between T cells and tumors has surged tremendously. In vitro coculture of tumor cells with autologous tumor-infiltrating lymphocytes (TIL) is a highly physiological and clinically relevant model to study functional T-cell responses that result from the array of activatory and inhibitory signals that naturally occur on the patient’s own tumor cells. This chapter describes a detailed protocol to set up a tumor-TIL coculture and assess ensuing functional T-cell responses, in order to establish the strength of tumor recognition by T cells and identify key determinants that govern this.

Key words

T cells Tumor-infiltrating lymphocytes (TIL) Tumor cells Immunotherapy Tumor immunology 


  1. 1.
    Yang Y (2015) Cancer immunotherapy: harnessing the immune system to battle cancer. J Clin Invest 125(9):3335–3337. Scholar
  2. 2.
    Pardoll DM (2012) The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12(4):252–264. Scholar
  3. 3.
    Prajapati K, Perez C, Rojas LBP, Burke B, Guevara-Patino JA (2018) Functions of NKG2D in CD8(+) T cells: an opportunity for immunotherapy. Cell Mol Immunol. Scholar
  4. 4.
    Blankenstein T, Coulie PG, Gilboa E, Jaffee EM (2012) The determinants of tumour immunogenicity. Nat Rev Cancer 12(4):307–313. Scholar
  5. 5.
    Betts MR, Brenchley JM, Price DA, De Rosa SC, Douek DC, Roederer M et al (2003) Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J Immunol Methods 281(1–2):65–78CrossRefGoogle Scholar
  6. 6.
    Schroder K, Hertzog PJ, Ravasi T, Hume DA (2004) Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol 75(2):163–189. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jeroen Melief
    • 1
    Email author
  • Stina Wickström
    • 1
  • Rolf Kiessling
    • 1
  • Yago Pico de Coaña
    • 1
  1. 1.Department of Oncology-PathologyKarolinska InstituteStockholmSweden

Personalised recommendations