Two c-type lectins from Venerupis philippinarum: Possible roles in immune recognition and opsonization

In the study, two c-type lectins were identified and characterized from the manila clam Venerupis philippinarum (designed as VpClec-1 and VpClec-2, respectively). Multiple alignments and phylogenetic analysis strongly suggested that they were new members of the c-type lectin superfamily. In normal tissue of clams, both VpClec-1 and VpClec-2 transcripts were highly expressed in the tissue of hepatopancreas. After Vibrio anguillarum challenge, the temporal expression of both VpClec-1 and VpClec-2 transcripts was up-regulated in the hemocytes of manila clams. The recombinant protein VpClec-1 (rVpClec-1) showed obvious binding activities to lipopolysaccharide (LPS), peptidoglycan (PGN), glucan and zymosan in vitro, while the recombinant protein VpClec-2 (rVpClec-2) could only bind LPS, glucan and zymosan. Coinciding with the PAMPs binding assay, both rVpClec-1 and rVpClec-2 displayed broad agglutination and antibacterial activities towards Vibrio harveyi, Vibrio splendidus, Vibrio anguillarum, Enterobacter cloacae and Aeromonas hydrophila. Moreover, the phagocytosis and encapsulation ability of hemocytes could be significantly enhanced by rVpClec-1 and rVpClec-2. Notably, the rVpClec-1 but not rVpClec-2 elicited a chemotactic response from hemocytes. All the results showed that VpClec-1 and VpClec-2 functioned as pattern recognition receptors (PRRs) with distinct recognition spectrum, and involved in the innate immune responses of manila clams.