New LEGENDplex™ Multi-Analyte Flow Assay Panels for Simultaneous Quantification of 13 Th Cytokines in Human and Mouse Samples

T helper cells (Th cells) play important roles in regulating the responses of the immune system. They secrete cytokines to stimulate various effector cells, such as cytotoxic T cells, B cells and macrophages. Accurate measurement of Th cytokine expression is critical to identify the corresponding Th cell responses and to develop in depth understanding of the immune responses. We have developed two multiplexed assay panels, i.e., the human and mouse Th panels, using fluorescence–encoded beads that are suitable for use on various flow cytometers. Both panels allow simultaneous quantification of 13 human or mouse cytokines, including interleukins (IL-2, 4, 5, 6, 9, 10, 13, 17A, 17F, 21, 22), IFN-γ and TNF-α, which are collectively secreted by Th1, Th2, Th9, Th17, Th22 or T follicular cells. Each antibody pair was carefully selected for assay specificity, sensitivity, accuracy and reproducibility. The assays provide higher detection sensitivity and broader dynamic ranges than traditional ELISA methods. The panels have been validated by detecting expected changes in biological samples. Further advantages include small sample volume, flexible assay configurations, and time- and cost-effectiveness. The Th cytokine panels can be used for serum, plasma, cell culture supernatant and other sample types, offering useful tools for biomedical research and drug discovery.