Functional characterization of two defensins, HlDFS1 and HlDFS2, from the hard tick Haemaphysalis longicornis.

Background: Ticks are second to mosquitoes as vectors of human arthropod-borne diseases. Ticks rely heavily on antimicrobial peptides (AMPs) to defend against microbes and defensins are major components of innate immunity in ticks.
Results: Two novel defensin genes, named HlDFS1 and HlDFS2, were identified from a cDNA library of the hard tick Haemaphysalis longicornis collected in southeast China. The peptides encoded by both genes shares typical features of type-2 arthropod defensin superfamily. The expressions of both genes increased in ticks during blood-feeding. The synthetic minimum functional peptides HlDFS1 and HlDFS2 showed broad spectrum antimicrobial activity against various Gram-positive and Gram-negative bacteria. Moreover, HlDFS1 and HlDFS2 exhibit bactericidal activity to some drug resistant bacteria. HlDFS1, but not HlDFS2, showed inhibitory activity against fungus Candida albicans. HlDFS1 and HlDFS2 had no significant hemolysis effect on human erythrocytes at low concentrations and did not impair mammalian cell survival. Finally, HlDFS1 and HlDFS2 significantly protected mice against lethal infection by Staphylococcus aureus and Micrococcus luteus.
Conclusions: HlDFS1 and HlDFS2 are two novel functional defensins from the hard tick Haemaphysalis longicornis. They showed bactericidal activity against various Gram-positive and Gram-negative bacteria and significantly protect mice against lethal bacterial infection. Thus, HlDFS1 and HlDFS2 can be introduced to the medical field as new drug candidates with antibacterial activity.