Molecular characterization, expression and functional analysis of goldfish (Carassius aurutus L.) interferon gamma

We report on the characterization and functional analysis of IFNgamma of the goldfish. Quantitative analysis indicated the highest expression of goldfish IFNgamma in the spleen, with lower expressions in brain, gill, kidney, heart, intestine and muscle. An increase in IFNgamma expression was observed in kidney leukocytes following stimulation with PHA, PolyI:C and during mixed leukocyte reaction. IFNgamma was expressed in different goldfish immune cell populations and this expression was upregulated in several of these populations following treatment of cells with recombinant goldfish TNFalpha-2. A recombinant form of goldfish IFNgamma (rgIFNgamma) was produced and functionally analyzed. The rgIFNgamma primed goldfish macrophages and neutrophils for enhanced respiratory burst responses and increased the phagocytic and nitric oxide responses of macrophages. Furthermore, treatment of goldfish macrophages with rgIFNgamma induced increased expression of several proinflammatory genes including TNFalpha-1, TNFalpha-2, IL-beta-1, IL-beta-2, IL-12-p35, IL-12-p40, RSAD-2 (=viparin), CXCL-8 (=IL-8), CCL-1, iNOS A and B and IFNgamma, and decreased the expression of TLR-3, while the expression of TGFbeta remained unchanged. Our results indicate that IFNgamma is a central proinflammatory cytokine of teleosts.