The proinflammatory cytokines TNF-α and IL-6 in lumpfish (Cyclopterus lumpus L.) -identification, molecular characterization, phylogeny and gene expression analyses

The proinflammatory cytokines TNF-alpha and IL-6 are important mediators of inflammatory reactions and orchestrators of the immune system in vertebrate. In this study, we have identified TNF-alpha and IL-6 in lumpfish, molecular characterized them at mRNA and gene level, performed homology modelling and measured their gene expression in different tissues and upon in vitro stimulation. A comprehensive phylogenetic analysis of TNF-alpha teleost sequences give novel insight into the TNF -alpha biology. Interestingly, we identified two isoforms of luIL-6. In normal tissue and leukocyte, the level of luTNF-alpha transcripts was higher than luIL-6. The expression pattern were parallel, except for brain, eye and gonad, and they displayed a similar induction pattern upon exposure to PAMPs, being most highly upregulated by flagellin. This is the first in-depth characterization of TNF and IL-6 in lumpfish. In recent years, lumpfish has become an important species for the aquaculture industry and establishment of qPCR-assays of luTNF-alpha and luIL-6 provide a valuable tool to measure effect of immune modulation, such as vaccination, microbiological disease and physiological trials. Lumpfish is also interesting for comparative studies as it represent a phylogenetic group that is poorly described immunologically.