Identification and initial functional characterization of lysosomal integral membrane protein type 2 (LIMP-2) in...

The immune system protects organism from external pathogens, this progress starts with the pathogen recognition by pattern recognition receptors (PRRs). As a group of PRRs, the class B scavenger receptors showed important roles in phagocytosis. Among three class B scavenger receptors, lysosomal integral membrane protein type 2 (LIMP-2) was reported to present in the limiting membranes of lysosomes and late endosomes, but its immune roles in teleost species are still limited in handful species. Here, we characterized LIMP-2 gene in turbot, and its expression patterns in mucosal barriers following different bacterial infection, as well as ligand binding activities to different microbial ligands and agglutination assay with different bacteria. In our results, one SmLIMP2 gene was identified with a 1,593 bp open reading frame (ORF). The multiple species comparison and phylogenetic analysis showed the closest relationship to Paralichthys olivaceus, the genomic structure analysis and syntenic analysis revealed the conservation of LIMP-2 during evolution. In tissue distribution analysis, SmLIMP-2 was expressed in all the examined turbot tissues, with the highest expression level in brain, and the lowest expression level in liver. In addition, SmLIMP-2 was significantly up-regulated in all the mucosal tissues (skin, gill and intestine) following Gram-negative bacteria Vibrio anguillarum infection, and was only up-regulated in gill following Gram-positive bacteria Streptococcus iniae challenge. Finally, the rSmLIMP-2 showed strong binding ability to all the examined microbial ligands, and strong agglutination with Escherichia coli, Staphylococcus aureus and V. anguillarum. Taken together, our results suggested SmLIMP-2 played important roles in fish immune response to bacterial infection. However, further functional studies should be carried out to better characterize its detailed roles in teleost immunity.