Cytokines are a family of secreted polypeptides that play a pivotal role in regulating immune responses, inflammation, development and tissue repairs. Since cytokines function in a complex regulatory network where one cytokine influences production of many other cytokines and vice versa, it has been found that profiling an array of cytokines in a given pathological condition is more biologically meaningful. Cytokine measurement is also important in determination of drug immunotoxicity during drug development. Drug induced toxicity to the immune system encompasses a variety of adverse effects, including suppression or enhancement of the immune response. Suppression of immune response can lead to decreased host resistance to infectious agents or tumor cells. Enhancing the immune response can exaggerate hypersensitivity or autoimmune disease.
The FlowCytomix Mouse Th1/Th2 10-Plex* cytokine assay kit is a multiplex fluorescent bead-based immunoassay for quantitative detection of Th1 and Th2 cytokines:GM-CSF, IFN-g, IL-1a, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17, TNF-a. Th1 cells produce IFN-g, TNF-a, IL-2, while Th2 cells produce IL-4, IL-5, IL-6 and IL-10. Th1 cytokines activate macrophages, NK cells, secretion of IgG subtype and cell mediated immunity. Th2 cytokines depress macrophage and cell mediated immunity. In this report, we describe a fully automated FlowCytomix bead-based assay procedure by employing the Biomek® FXP Laboratory Automation Workstation. Analyses of the assayed beads were conducted on either the Cell Lab Quanta™ SC MPL or the FC 500 MPL Flow Cytometry System. The results demonstrated that similar assay sensitivity and reproducibility with FlowCytomix Mouse Th1/Th2 10-Plex kit were generated on both FC 500 MPL and Quanta SC MPL. Automation of this multiplexed flow cytometry-based assay dramatically increases the sample throughput and simplifies the sample preparation process. |
