University Hospital Herlev 1 article published in JoVE Immunology and Infection Real-time Monitoring of Mitochondrial Respiration in Cytokine-differentiated Human Primary T Cells Kasper Mølgaard*1, Anne Rahbech*1, Özcan Met1, Inge Marie Svane1, Per thor Straten1,3, Claus Desler2, Marlies J. W. Peeters1 1National Center for Cancer Immune Therapy, Department of Oncology, University Hospital Herlev, 2Department of Cellular and Molecular Medicine, Center for Healthy Aging, University of Copenhagen, 3Department of Immunology and Microbiology, Inflammation and Cancer Group, University of Copenhagen Metabolic adaptation is fundamental for T cells as it dictates differentiation, persistence, and cytotoxicity. Here, an optimized protocol for monitoring mitochondrial respiration in ex vivo cytokine-differentiated human primary T cells is presented.