Pathogen recognition and mechanisms in Atlantic cod (Gadus morhua) head kidney cells: Bacteria (LPS) and virus (poly I:C) signals through different pathways and affect distinct genes

Abstract

Understanding pathogen recognition and mechanisms in Atlantic cod are of significant importance for both basic research on wild populations and health management in aquaculture. A microarray approach was utilized to search for effects of viral (polyinosinic acid:polycytidylic acid), bacterial (lipopolysaccharide) and polyclonal activator (phytohaemoagglutinin) stress in Atlantic cod head kidney cells. LPS cell activation increased mRNA expression of interleukin 8; interleukin-1β; cyclooxygenase 2; leukocyte derived chemotaxin 2; carboxyl-esterase 2 and environmental biomarker cytochrome P450 1A. Mitogen activated protein kinase p38 and cathepsin F were down regulated by LPS. The antiviral responses induced by double stranded RNA clearly increased transcription of Toll like receptor 3 and interferon stimulating gene 15. The phytohaemoagglutinin response seemed to be more non-specific. Special for the phytohaemoagglutinin induction was the increase in major histocompatibility complex class I. CC chemokine type 2 mRNA expression was increased by phytohaemoagglutinin, lipopolysaccharide and polyinosinic acid:polycytidylic acid, while mitogen activated protein kinase p38 and leukocyte derived chemotaxin 2 were down regulated by phytohaemoagglutinin. Oxidative stress related genes like catalase and glutaredoxin and the anti-apoptotic gene Bcl-2 showed no transcriptional changes compared to control in any of the treatments. Eicosanoids like prostaglandin 2, leukotriene B4 and B5 were constitutively produced by cod head kidney cells in vitro. The most remarkable feature of eicosanoid secretion is the higher production of leukotrienes against prostaglandins, indicating that the lipooxygenase pathway is preferred over the cyclooxygenase pathway. Although there were no significant differences in eicosanoid secretion between the groups, polyinosinic acid:polycytidylic acid showed a clear tendency to increase the levels of leukotriene B4 and B5. This study reveals distinct signatures of bacteria and virus transcriptional responses in cod head kidney cells. In addition, the novel finding that cytochrome P450 1A was upregulated during the antibacterial response indicates a connection between immunity and aryl hydrocarbon receptor activation in Atlantic cod.