Abstract 1624: Immune cell killing assay for measuring the effects of immunomodulating agents in lung cancer cells in vitro

Abstract Direct effects of cancer therapeutics on immune cell activity can be challenging to determine on system level in vivo. Therefore, it is important to develop and discover novel ways to study the effects of immune cells on cancer cells using also in vitro assays. Our aim was to validate an in vitro immune cell killing assay where activated peripheral blood mononuclear cells (PBMCs) target lung cancer cells and induce apoptosis. A549 human lung carcinoma cells (ATCC) were used as target cells in the co-culture and were plated into 96-well plates. 24 hours later, human PBMCs (Lonza) were added to the culture as effector cells and activated with anti-CD3 and IL2. Apoptosis was detected using IncuCyte Annexin V reagent, which was added to the cells at the same time with immune cell activators. Cell proliferation was monitored with a live cell imager IncuCyte S3 (Sartorius) for three to four days before terminating the cultures. Cell viability was determined by CellTiter Glo (Promega) at the end of the study. Treatment of cells with anti-CD3 was able to activate immune cells alone but the effects were observed with lower test concentrations in combination with IL-2. Therefore, IL-2 potentiates the effect of anti-CD3 on immune cell activation and decreases cancer cell viability in co-culture. The immune cell killing assay based on live cell imaging can be used for studying the effects of immunomodulating agents on cancer cell viability and to evaluate the activity of immune cells in the presence of therapeutics. Citation Format: Jenni H. Mäki-Jouppila, Jukka P. Rissanen, Katja M. Fagerlund. Immune cell killing assay for measuring the effects of immunomodulating agents in lung cancer cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1624.