Changes in immune gene expression and resistance to bacterial infection in lobster ( Homarus gammarus) post-larval stage VI following acute or chronic exposure to immune stimulating compounds

Abstract

Real-time PCR was used to measure changes in transcript abundance of genes encoding important immune proteins, namely prophenoloxidase ( proPO gene), β-1,3-glucan binding protein ( βGBP gene) and a 12.2 kDa antimicrobial peptide ( amp gene) in post-larval stage VI (PLVI) juveniles of the European lobster, Homarus gammarus. Gene expression was studied in both healthy PLVI and following single or repeat exposure to a range of compounds claimed to induce immune reactivity. A single acute (3-h) exposure to any of the tested stimulants did not produce a significant increase in expression of either the proPO or βGBP genes, measured 6 h after stimulation. However, there were a small sub-group of positive responders, identified mainly from βGBP expression, within the experimental groups stimulated with either a β-1,3-glucan or an alginate. There was also no significant increase in the expression of any of the three genes tested 24 h after repeated weekly (3-h) exposures to a either the β-1,3-glucan or the alginate over the longer (36-day) period. The results do show that amp is expressed at an extremely high level compared to proPO or βGBP in healthy animals and a significant correlation was found between the expression of proPO and both βGBP and amp, irrespective of whether or not the larvae were stimulated. None of the immune stimulated compounds improved survival of PLVI challenged with the opportunistic pathogen, Listonella anguillarum, or the lobster pathogen, Aerococcus viridans var. homari. Thus, we found no evidence to support recent claims that immunity and disease resistance can be primed or promoted within a given population of crustaceans or that these animals exhibit functional immune memory to some soluble immune elicitors.