Interleukin-36β inhibits CD4+CD25+ regulatory T cells by activating endoplasmic reticulum-phagy in septic mice

CD4+CD25+ regulatory T cells (Tregs) contribute to the pathogenesis of sepsis-induced immunosuppression. We have identified interleukin (IL)-36β as a critical cytokine regulating CD4+CD25+ Treg activity. This study aimed to further investigate the underlying mechanism of IL-36β-triggered responses in murine CD4+CD25+ Tregs in presence of lipolysaccharide (LPS) and in a mouse model of sepsis induced by cecal and puncture (CLP). Following LPS exposure, ER-phagy activity increased, peaked at 12 h, and then markedly declined. Furthermore, we observed that IL-36β could activate ER-phagy of CD4+CD25+ Tregs under LPS challenge. Mechanistic investigations revealed the critical involvement of the endoplasmic reticulum (ER) stress-related protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK)-activating transcription factor 4 (ATF4) signaling axis in IL-36β-induced ER-phagy. Moreover, IL-36β knockout (IL-36β-/-) strongly dampened ER-phagy and PERK-ATF4 signaling under LPS stimulation compared to the wild-type group. IL-36β-elicited effects on CD4+CD25+ Tregs were significantly abrogated by FAM134B (the ER-phagy-specific receptor) knockout or salubrinal (a specific inhibitor of the PERK-ATF4 pathway). In addition, IL-36β was potent in diminishing serum levels of creatinine (Cr), aspartate transaminase (AST), and alanine transaminase (ALT) and attenuated histopathologic alterations in the liver, kidneys, and lungs of CLP mice. Importantly, the absence of IL-36β notably aggravated the survival rate of septic mice, indicating a beneficial role in septic prognosis. IL-36β can down-regulate the immune activity of CD4+CD25+ Tregs via ER-phagy induction. Our study might provide novel targets for therapeutic strategies to prevent the development of sepsis.