Involvement of galectin-9 from koi carp (Cyprinus carpio) in the immune response against Aeromonas veronii infection

Galectins are beta-galactoside sugar binding proteins which function as important pattern recognition receptors (PRRs) in innate immunity. Here, we identified a galectin-9 gene from koi carp (Cyprinus carpio), named kGal-9. The ORF of kGal-9 is 963 bp in length, which encodes a polypeptide of 320 amino acids without either signal peptide. The predicted molecular weight is 36.25 kDa, and the isoelectric point is 8.3. Multiple sequence alignment showed that the putative kGal-9 contains two carbohydrate recognition domains (CRD), which are conserved in Galectin-9s. The phylogenetic tree showed that kGal-9 clustered to Galectin-9s from other teleosts, and shared the highest identity of 87.5% with Qihe crucian (Carassius auratus). kGal-9 mRNA was abundant in head kidney, gills, and gut, but low in liver and muscle. Further, the expression level of kGal-9 in the head kidney and liver increased significantly after Aeromonas veronii (abbreviated A.v) infection. Unexpectedly, kGal-9 showed a remarkable downregulation in the spleen at various time points post A.v infection. Intramuscular injection of pckGal-9 not merely reduced the bacterial load of spleen tissue, but also improved the survival rate of koi carp post A.v challenge. Besides, administration of pckGal-9 stimulated the expression of several immuno-related genes including proinflammatory cytokines (IL-1 beta, IL-6), anti-inflammatory cytokine (IL-10), complement components (C4, C9), with fluctuation in spleen and head kidney. Taken together, the obtained results suggest that kGal-9 occupies an important role in innate immunity and defense against bacterial infection in koi carp.