Multiplex analysis of inflammatory cytokines from primary human macrophages using a FLUOstar Omega
During the autoimmune disease Rheumathoid Arthritis immune competent cells such as macrophages and T-cells accumulate in joints. There they secrete the cytokines IL6, IL10 and TNFα. These activate the transcription factor NFκB which promotes inflammation.
Using the FLUOstar® Omega multi-mode plate reader cytokine secretion and NFκB-induction were measured in macrophages upon various stimuli. The cytokines IL6 and TNFα were quantified in the culture supernatant by absorbance-based ELISAs. A luciferase reporter assay was used to detect transcriptional activation of NFκB. Lipopolysaccharide stimulated macrophages showed a higher luminescence, indicative of NFκB activation, than their non-stimulated controls.
The FLUOstar® Omega analyses multiple cytokines on the same plate. We show that human primary macrophages can be cultured in 384-well plates without any changes in morphology or cytokine expression. Reducing the number of cells for each assay without altering the sensitivity now opens possibilities to do screening assays on primary cells to look for new targets in autoimmune diseases.