Combating Pathogenic Immune Evasion: Sialidase-Activated Thermally Delayed Fluorescence for Probing and Modulating Host-Pathogen Interactions

Innate immunity represents the primary defense against invasive pathogens with phagocytosis playing a central role in host defense and mediating immune and inflammatory responses. However, pathogens such as Clostridium perfringens have developed strategies to overcome phagocytic clearance. Developing molecular tools to identify and target key factors in pathogenic immune evasion can deepen our understanding of host-pathogen interactions and aid in exploring novel therapeutic strategies. As a key enzyme in the sialylation process of C. perfringens, the virulence factor sialidase is a potential target for investigating pathogenic immune evasion. Herein, a "turn-on" thermally activated delayed fluorescent probe SA-HBT-PXZ is developed as a highly selective and sensitive sialidase sensor, enabling time-resolved fluorescence imaging of C. perfringens in live bacterial cells, tissue sections, and even infected mice. Furthermore, SA-HBT-PXZ is successfully employed to screen sialidase inhibitors based on prompt and delayed fluorescence emissions. The identified lead compounds effectively inhibit the activity of sialidases from C. perfringens, leading to an increased level of differentiation of macrophages into the M1 subtype. This, in turn, enhances the phagocytosis of C. perfringens and ultimately suppresses the immune escape of the bacteria. Our study provides a potential target and lead compounds for novel therapeutic strategies against C. perfringens infections.