Expression of Interleukin-1β protein in vitro,exvivo and in vivo salmonid models

Abstract

Interleukin-1β (IL-1β) is one of the first cytokines expressed during immune responses, and its levels are affected by many factors, including stress. To date, it has only been possible to measure IL-1β transcript (mRNA) expression quantitatively in fish using qPCR. This is because previous studies that measured IL-1β protein concentrations in these taxa used western blotting, which only provides qualitative data. To advance our knowledge of fish IL-1β biology, and because post-translational processing plays a critical role in the activation of this molecule, we developed a quantitative enzyme-linked immunosorbent assay (ELISA) to accurately measure the concentration of IL-1β protein in several cell cultures and in vivo in salmonids. We compared changes in IL-1β protein levels to the expression of its mRNA. The developed ELISA was quite sensitive and has a detection limit of 12.5 pg/mL. The tools developed, and information generated through this research, will allow for a more accurate and complete understanding of IL-1β's role in the immune response of salmonids.The assay described here has the potential to significantly advance our ability to assess fish health and immune status.

Keywords: Cell culture; Diagnostic assay; Immunostimulation; Interleukin-1β; Natural infection; Quantitative ELISA.