Detection of glial interleukin-6 release: Development of a novel interleukin-6 Elisa
The development of an interleukin-6 (IL-6) enzyme-linked immunosorbent (ELISA) assay resulted in a detection range of 1 ng/m1 to 10 ng/mL IL-6. The limit of detection (LOD) for IL-6 was 0.25 ng/mL. Troubleshooting techniques were attempted including matrix effects, non-specific binding, and coating buffer pH differences. In comparison to the ELISA, the LOD for IL-6 using the 7TD1 bioassay was 2--5 pg/mL. Therefore the 7TD1 bioassay was implemented for the experiments described in this study. IL-6 is a cytokine known to stimulate B-cell differentiation and to activate T-cells. IL-6 is elevated in inflammatory and neurodegenerative diseases. Interleukin-1beta (IL-1beta) stimulates IL-6 release from glial cells. Catecholamines and IL-1beta synergistically release IL-6 from glial cells. In the rat C6 glioma cell line, the role of indoleamines was examined alone and in combination with IL-1beta resulting in no synergistic effects on IL-6 release. Isoproterenol synergistically acted with IL-1beta on stimulating IL-6 release. IL-1 receptor antagonist decreased the IL-1beta stimulation of IL-6. These results demonstrate that the catecholamines and IL-1beta play a role in the regulation of IL-6 expression.
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