Toll and IMD Pathways Synergistically Activate an Innate Immune Response in <i>Drosophila melanogaster</i>

TLDR

In Drosophila melanogaster, antimicrobial peptide gene expression is controlled by the Toll and IMD signaling pathways, which are thought to act independently but can jointly activate some target genes, possibly through cross‑regulation. This study aims to demonstrate that Toll and IMD pathways interact synergistically via cross‑regulation. RNA interference and promoter analyses reveal that cooperation among distinct NF‑κB‑related transcription factors mediates this synergy. Co‑stimulation with Spätzle and gram‑negative peptidoglycan, or constitutive activation of both pathways, synergistically induces genes such as Drosomycin, Diptericin, and AttacinA, illustrating how distinct pattern‑recognition receptors translate into a broad‑spectrum innate immune response.

Abstract

The inducible expression of antimicrobial peptide genes in Drosophila melanogaster is regulated by the conserved Toll and peptidoglycan recognition protein LC/immune deficiency (PGRP-LC/IMD) signaling pathways. It has been proposed that the two pathways have independent functions and mediate the specificity of innate immune responses towards different microorganisms. Scattered evidence also suggests that some antimicrobial target genes can be activated by both Toll and IMD, albeit to different extents. This dual activation can be mediated by independent stimulation or by cross-regulation of the two pathways. We show in this report that the Toll and IMD pathways can interact synergistically, demonstrating that cross-regulation occurs. The presence of Spätzle (the Toll ligand) and gram-negative peptidoglycan (the PGRP-LC ligand) together caused synergistic activation of representative target genes of the two pathways, including Drosomycin, Diptericin, and AttacinA. Constitutive activation of Toll and PGRP-LC/IMD could mimic the synergistic stimulation. RNA interference assays and promoter analyses demonstrate that cooperation of different NF-κB-related transcription factors mediates the synergy. These results illustrate how specific ligand binding by separate upstream pattern recognition receptors can be translated into a broad-spectrum host response, a hallmark of innate immunity.