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Ultrapure LPS induces inflammatory and antibacterial responses attenuated in vitro by exogenous sera in Atlantic cod and Atlantic salmon

Seppola, M;Mikkelsen, H;Johansen, A;Steiro, K;Myrnes, B;Nilsen, IW;

Phagocyte recognition of lipopolysaccharide (LPS) is an early key event for triggering the host innate immune response necessary for clearance of invading bacteria. The ability of fishes to recognise LPS has been questioned as contradictory results have been presented. We show here that monocyte/macrophage cultures from Atlantic cod (Gadus morhua) and Atlantic salmon (Salmo salar) respond with an increased expression of inflammatory and antibacterial genes to both crude and ultrapure Escherichia coli LPS. Crude LPS produces higher induction than the ultrapure LPS type in both species in vitro as well as in vivo in cod injected with LPS. Crude LPS gave, in contrast to ultrapure LPS, an additional weak up-regulation of antiviral genes in salmon macrophages, most likely because of contaminants in the LPS preparation. Increased levels of chicken (c)-type lysozyme transcripts and enzyme activity were measured in salmon macrophages following ultrapure LPS stimulation demonstrating not only increased transcription but also translation. Simultaneous use and even pre-treatment with bovine sera suppressed the LPS-induced expression thereby reflecting the presence of transcription inhibitory components in sera. Together, these findings show that both cod and salmon recognise LPS per se and that the observed induction is highly dependent on the absence of sera.