Molecular Characterization of an IL-1β Gene from Ayu, Plecoglossus Altivelis
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IL-1β plays a crucial role as a prototypical proinflammatory cytokine in immune responses and has been shown to affect macrophage functions. However, the effects of putative IL-1β homologs on fish macrophages are still less known. Here, we cloned the full-length cDNA sequence of IL-1β (aIL-1β) gene from ayu, Plecoglossus altivelis. Phylogenetic analysis indicated that aIL-1β was closest to that of Atlantic salmon (Salmo salar). Real-time quantitative PCR (RT-qPCR) revealed that aIL-1β transcript was mainly expressed in spleen, head kidney and gill, and dramatically increased in various tissues after Listonella anguillarum infection. Subsequently, aIL-1β was prokaryotic expressed and purified to prepare anti-aIL-1β antibody. After L. anguillarum challenge, the aIL-1β mRNA and protein levels were significantly up-regulated in ayu monocytes/macrophages. Moreover, aIL-1β neutralization did not change phagocytic capability, but reduced bacterial killing capability in ayu head kidney-derived monocytes/macrophages. Therefore, aIL-1β may play an important role in immune response of ayu, especially, contributing to bacterial killing of monocytes/macrophages.
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