Zebrafish larvae as a powerful model to dissect protective innate immunity in response to Legionella pneumophila infection

The zebrafish has become a powerful model organism to study host-pathogen interactions. Here, we developed a zebrafish model of Legionella pneumophila infection to dissect innate immune responses. We show that L. pneumophila cause zebrafish larvae death in a dose dependent manner, and that macrophages are the first line of defence, with neutrophils cooperating to clear the infection. When either macrophages or neutrophils are depleted, these "immunocompromised" larvae become lethally sensitive to L. pneumophila similar to what is known for humans that develop pneumonia. Also as observed in human infections, the adaptor signalling molecule Myd88 is not required to control disease in the larvae. Furthermore, proinflammatory cytokine genes il1{beta} and tnf were upregulated during infection, recapitulating key immune responses seen in human infection. Strikingly, we uncovered a previously undescribed infection phenotype in zebrafish larvae, whereby bloodborne, wild type L. pneumophila invade and grow in the larval yolk region, a phenotype not observed with a type IV secretion system deficient mutant that cannot translocate effectors into its host cell. Thus, zebrafish larva represents an innovative L. pneumophila infection model that that on one hand mimics important aspects of the human immune response to L. pneumophila infection and that on the other hand will allow to elucidate the mechanisms by which type IV secretion effectors allow L. pneumophila to cross membranes and to obtain nutrients from nutrient rich environments.