2,6-dichloro-1,4-benzoquinone (DCBQ), as an emerging water disinfection byproducts (DBPs), has posed potential risks via the digestion system. However, little is known about the toxicity of DCBQ on the gut microbiome, which plays a critical role on human health. This study has comprehensively investigated the impact of DCBQ on the intestinal microbiome, metabolic functions, and immunity after the mice orally exposure to DCBQ at the concentration of 31.25, 62.5 and 125 mg/kg body weight for 28 days. Our results indicated that DCBQ exposure has perturbed the balance between T helper (Th) 1 mediated pro-inflammatory response and Th2 mediated anti-inflammatory response in mice, especially inducing the activation of immune system toward a Th2 response. DCBQ group has induced gut microbiota dysbiosis, and at phylum level, Proteobacteria was relatively less abundant compared with that in the control group. Furthermore, DCBQ exposure has dramatically perturbed metabolites profiles which were involved in 28 metabolic pathways, such as amino acids biosynthesis and metabolism, lipid metabolism. In particular, the altered gut microbiota showed strong correlations with both the altered metabolites and the altered immunological variables after DCBQ exposure. This study provides evidence on the adverse effects and mechanisms of water disinfection byproduct DCBQ through the interaction of immune-microbiome-metabolome, highlighting the importance to assess DBPs-associated risks.