Abstract 4855: Metabolic and functional dynamics of T cell activation: bioluminescent assays for immune monitoring and modulation

Abstract Advancing our understanding of T cell activation and its regulatory pathways is critical for developing effective cancer immunotherapies. This study characterizes T cell health and functional dynamics under diverse activation conditions, highlighting key molecular mechanisms that influence immune responses within the tumor microenvironment. We investigate mTOR and AMPK signaling pathways, as well as the metabolic balance between glycolysis and oxidative phosphorylation, which are integral to T cell function in immunotherapeutic contexts. Using selective inhibitors, we identify modulators that regulate cytokine secretion and T cell metabolic states, revealing media composition as a significant variable affecting immune function. Our methodology leverages a suite of bioluminescent Lumit and Metabolite-Glo assays for sensitive quantification of cytokines and metabolites, enabling precise, non-invasive monitoring while preserving cell integrity. This approach is scalable and adaptable for analyzing diverse activation conditions and high-throughput screening applications. Additionally, our findings correlate these metabolic and functional profiles with cell surface markers assessed by traditional methods, providing a comprehensive framework for immune monitoring. These insights underscore the importance of metabolic and signaling interplay in T cell activation, offering a robust platform for studying immune dynamics and uncovering novel immunomodulatory targets. Our work supports the development of precision strategies to modulate T cell activity and optimize immune responses in cancer therapy. Citation Format: Kim Haupt, Kayla Sylvester, Tony Lauer, Natasha Karassina, Jolanta Vidugiriene. Metabolic and functional dynamics of T cell activation: bioluminescent assays for immune monitoring and modulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4855.