Damage signal induced by Bacillus thuringiensis infection triggers immune responses via a DAMP molecule in lepidopteran insect, Spodoptera exigua.

Insect immunity defends the infection of an insect pathogenic bacterium, Bacillus thuringiensis (Bt). However, it was not clear on the recognition of Bt infection by the insect immune system. This study tested a physiological function of dorsal switch protein 1 (DSP1) in the Bt infection. DSP1 is classified into HMGB1-like damage-associated molecular pattern (DAMP) in insects. Upon Bt infection in a lepidopteran Spodoptera exigua, DSP1 was released from the nuclei of the midgut epithelium and activated immune responses. For this DSP1 release, a functional binding between Bt and its receptors on the midgut epithelium was required because any RNA interference (RNAi) treatments of Bt receptor (cadherin or ABCC) prevented the DSP1 release and became susceptible to the bacterial infection. The DSP1 release was required for the gene induction of Repat33, which is a member of response to pathogen gene family and its gene product mediated cellular and humoral immune responses against pathogen infection in S. exigua. The released DSP1 activated phospholipase A (PLA) to produce eicosanoids, which induced the Repat33 expression because a hemocoelic injection of a recombinant DSP1 induced the Repat33 expression without Bt infection. However, any inhibition of PLA activity impaired the DAMP signaling between DSP1 and Repat33. DSP1 also up-regulated two other immune mediators, nitric oxide (NO) and a cytokine called plasmatocyte-spreading peptide (PSP). Either NO or PSP activated PLA to up-regulate Repat33 expression. These results suggest that Bt infection of the insect midgut generates a DAMP signal via DSP1 release, which turns on NO or the cytokine-PLA-Repat33 immune signaling pathway.