LAUSR

ABSTRACT Although aluminum salts (Alum) have been extensively used in human vaccination for decades, its mechanism of action is controversial. In fish, the use of Alum as a vaccine adjuvant is scarce and there are no studies aimed at identifying its mechanism of action. In the present study we report that Alum is a powerful adjuvant in the gilthead seabream (Sparus aurata L., Sparidae) and the European seabass (Dicentrarchus labrax L. Moronidae). Thus, Alum increased the specific antibody titers to the model antigen keyhole limpet hemocyanin as the commonly used Freund's adjuvant did in both species. In addition, both adjuvants were able to increase the transcript levels of the gene encoding the major pro‐inflammatory mediator interleukin‐1&bgr; (Il1b). Strikingly, however, Alum failed to promote Il1b release by seabream leukocytes and even impaired Il1b induction, processing and release in macrophages. However, it increased NADPH oxidase‐dependent reactive oxygen species (ROS) production in gilthead seabream leukocytes and purified granulocytes. In addition, Alum promoted gilthead seabream leukocyte death independently of ROS production and caspases, suggesting that damage‐associated molecular patterns release from dying cells mediate Alum adjuvant activity. Our results pave the way for future studies aimed at investigating the relevance of danger signals generated by Alum in vivo on its adjuvant activity in order to increase our understanding of the mechanisms of action of Alum in fish vaccines and to help in the design of new adjuvants for aquaculture. HighlightsAlum increases the specific antibody titers to KLH in gilthead seabream and European seabass.Alum and Freund's adjuvants induce the expression of gene coding for pro‐inflammatory interleukin‐1&bgr;.Alum fails to promote interleukin‐1&bgr; release by seabream leukocytes.Alum increases NADPH oxidase‐dependent reactive oxygen species (ROS) production and cell death.Damage‐associated molecular patterns release from dying cells may mediate Alum adjuvant activity.