Evaluating functional anti-tumor T cell responses in autologous 2D co-cultures using flow cytometry

Immune surveillance by T cells is a key determinant of cancer progression, and T cell-based immunotherapies have shown great promise as a treatment modality for cancer patients. As such, physiologically relevant methods to evaluate interactions between T cells and tumor cells are of particular interest. In vitro systems that enable the culture patient-derived tumor cells in the presence of autologous tumor-infiltrating lymphocytes (TIL) serve as an invaluable tool to understand the basic biological role of T cells in cancer and how their functioning might be modulated to gain therapeutic benefit. Hence, this chapter describes a flow cytometry-based approach to assess TIL activation by exposure to autologous tumor cells in culture. In particular, the chapter will focus on ways to assess the capacity of cytotoxic lymphocytes (CTL) to degranulate and secrete pro-inflammatory cytokines in such culture systems.