The innate immune gene Relish and Caudal jointly contribute to the gut immune homeostasis by regulating antimicrobial peptides in Galleria mellonella.

Gut microbiota modulates various physiologic processes in insects, such as nutrition, metabolic homeostasis, and pathogen exclusion. Maintaining a normal microbiome is an essential element of the gut homeostasis, requiring an extensive network of regulatory immune responses. The molecular mechanisms driving these various effects and the events leading to the establishment of a normal microbiota in insects are still largely unknown. In this study, the NF-kB (IMD and Toll) signaling pathways in the gut of Galleria mellonella and their roles in the regulation of its gut microbes were assessed. For this, the transcript levels of the IMD pathway (Imd and Relish) and the Toll pathway (Spätzle and Dif/Dorsal) genes were analyzed and the results showed that all the genes were expressed in the gut of G. mellonella. Silencing of Relish resulted in reduced expression levels of the IMD pathway genes and antimicrobial peptides (AMPs) followed by overpopulation of gut bacteria. Antibiotics-treated larvae showed lower expression levels of the IMD and Toll pathway genes followed by lower AMPs expression levels. The expression level of caudal decreased in the antibiotics-treated larvae compared with the controls. Together, these data suggest that the IMD and Toll pathways are active in the gut of G. mellonella. The IMD pathway gene, relish functions in the regulation of gut microbes in this insect model.