Abstract 2759: Immune cell killing assay for analyzing the effects of immunomodulating agentsin vitro
2020
Immuno-oncology has emerged as one of the most promising new approaches in cancer drug development. Greater cancer cell specificity of treatments can be achieved when the immune cells of a patient are used against tumors. Therefore, it is important to study the effects of immune cells on cancer cells using different models in vitro and in vivo. Our aim was to validate an in vitro T cell killing assay where activated peripheral blood mononuclear cells (PBMCs) target different cancer cells and increase their apoptosis. MCF-7 human breast carcinoma cells (ATCC) were used as target cells and were seeded on 96-well plates. 24 hours later, human PBMCs (Lonza) were added to the culture as effector cells. Simultaneously, the PBMCs were activated with 100 ng/ml anti-CD3 and 10 ng/ml IL2. Apoptosis was detected using IncuCyte Annexin V reagent, which was introduced to the cells at the same time with activators. Cell growth, proliferation and apoptosis were monitored with a live cell imager IncuCyte S3 (Sartorius) for three days. Cell viability was determined by CellTiter Glo (Promega) at the end of the study. The target-to-effector ratio of 1:5 and 1:10 both showed an increase in apoptosis of MCF-7 cells in the wells with activated PBMCs. This was detected with a confluence parameter (percentage) of IncuCyte S3. The T cell killing assay can be utilized for studying cancer therapeutics based on immune-oncology (e.g. pembrolizumab and atezolizumab) and to evaluate the potential of immune cells in killing cancer cells in the presence of drugs affecting the target-effector interaction. Citation Format: Jenni H. Maki-Jouppila, Jussi M. Halleen, Katja M. Fagerlund. Immune cell killing assay for analyzing the effects of immunomodulating agents in vitro [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2759.
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