Layers of immunity: Deconstructing the Drosophila effector response

The host innate immune response relies on the cooperation of multiple defense modules. In insects and other arthropods, which have only innate immune mechanisms, four main immune-specific modules contribute to defense against microbial invaders: the Toll pathway, the Imd pathway, the melanization response, and phagocytosis by plasmatocytes. Our present understanding of their relative importance remains fragmented, as their contribution to host defense has never been simultaneously assessed across a large panel of pathogens. Here, we have taken advantage of newly-described immune mutants in a controlled genetic background to systematically delete these four immune modules individually, in pairs, or all four simultaneously. Surprisingly, flies simultaneously deficient in all four immune modules are viable, homozygous fertile, and display no overt morphological defects, suggesting these immune mechanisms are not strictly required for organismal development. With this new set of lines, we assessed the individual and collective contribution of each module to host defense against five viruses, three fungi, eight Gram-positive bacteria, and eight Gram-negative bacteria. Our findings show that these four modules largely function independently or additively in host defense, although synergistic effects can occur for select pairs of modules. Our study confirmed the importance of the Imd pathway against Gram-negative bacteria and the Toll pathway against Gram-positive bacteria and fungi, largely via the induction of effectors such as antimicrobial peptides (AMPs) and Bomanins (Bom), but also reveals an important role of melanization against viruses, and a contribution of phagocytosis against various germs. Additionally, by examining microbial load kinetics in different mutants, we provide insight into how these modules contribute to tolerance or resistance against specific microbes. Our study provides insights into the architecture of the Drosophila immune system, revealing differential requirements of immune modules according to each pathogen. The set of immune deficient lines provided here offers tools to better assess the role of these immune modules in host defense.