Immune priming in the insect gut: a dynamic response revealed by ultrastructural and transcriptomic changes

Abstract

Research on forms of memory in innate immune systems has recently gained momentum with the study of trained immunity in vertebrates and immune priming in invertebrates. Immune priming provides protection against previously encountered pathogens. However, causes and mechanisms of immune priming are still not well understood in most organisms. In this work, we combine RNA sequencing with transmission electron microscopy to investigate the dynamic processes during priming in the gut of a well-established model for oral immune priming, consisting of the host Tribolium castaneum and its entomopathogen Bacillus thuringiensis tenebrionis (Btt). We show that priming with specific, pathogen-derived virulence-relevant factors causes damage in the gut of T. castaneum larvae, which leads to an early physiological stress response as well as the upregulation of a specific set of immune genes. This response diminishes over time yet enables the gut to upregulate genes known to interfere with Btt virulence when T. castaneum larvae are later exposed to infectious Btt spores. The identification of these processes increases our understanding of immune priming as a dynamic process where cellular responses in concert with specific gene regulation prepare the gut tissue and thereby enable more efficient protection against infection. Such work can further help us understand the origin and mechanism of innate immune memory. Author summaryInvertebrate animals do not possess a classical adaptive immune system. And yet, many of these animals show forms of immune memory, collectively called immune priming. In this work, we investigate the causes of oral immune priming in the red flour beetle Tribolium castaneum in response to its bacterial entomopathogen Bacillus thuringiensis tenebrionis. Primary exposure to Btt spent culture media supernatants enables the larvae of T. castaneum to better withstand a subsequent infection with infectious Btt spores. We show that exposing T. castaneum larvae to those supernatants leads to damage in the gut resulting in a strong stress and immune response early after priming and a targeted up-regulation of beneficial genes upon the secondary re-encounter with Btt. These results suggest fundamental stress responses can be involved in innate immune memory phenomena, which has implications in translational research across species.