Listeria monocytogenes(Lm) is a facultative, intracellular Gram-positive pathogenic bacterium that causes sepsis, a condition characterized by persistent excessive inflammation and organ dysfunction. However, the pathogenesis ofLm-induced sepsis is unknown. In this research, we discovered that TRIM32 is required for innate immune regulation duringLminfection.Trim32deficiency remarkably reduced bacteremia and proinflammatory cytokine secretion in mice with severeLminfection, preventing sepsis.Trim32−/−mice had a lower bacterial burden afterLminfection and survived significantly longer than wild-type (WT) mice, as well as lower serum levels of inflammatory cytokines TNF-α, IL-6, IL-18, IL-12p70, IFN-β, and IFN-γ at 1 day post infection (dpi) compared to WT mice. On the other hand, the chemokines CXCL1, CCL2, CCL7, and CCL5 were enhanced at 3 dpi inTrim32−/−mice than WT mice, reflecting increased recruitment of neutrophils and macrophages. Furthermore,Trim32−/−mice had higher levels of macrophage-associated iNOS to killLm. Collectively, our findings suggest that TRIM32 reduces innate immune cells recruitment andLmkilling capabilities via iNOS production.Graphical AbstractSupplementary InformationThe online version contains supplementary material available at 10.1186/s13099-023-00558-9.