Outer membrane vesicles (OMVs) derived from Vibrio species serve as important delivery vehicles for effector molecules and play critical roles in host–pathogen interactions. However, the systemic immune response patterns elicited by Vibrio-derived OMVs in the Pacific white shrimp (Litopenaeus vannamei) remain poorly characterized. In this study, we investigated the immune response features of L. vannamei following stimulation with OMVs derived from two Vibrio parahaemolyticus strains responsible for acute hepatopancreas necrosis disease (AHPND) and translucent postlarvae disease (TPD), respectively. In vivo imaging and hemocyte uptake assays revealed that OMVs preferentially accumulated in the hepatopancreas and intestine, two major immune-related tissues, accompanied by pronounced hemocyte-mediated active internalization. Following in vivo injection, shrimp exhibited marked immune activation, as evidenced by alterations in total hemocyte count, redistribution of hemocyte subpopulations, and elevated reactive oxygen species (ROS) levels, indicating a systemically activated immune state. At the transcriptional level, the hepatopancreas and intestine displayed distinct tissue-specific immune response patterns to the two OMVs, which were accompanied by coordinated changes in antioxidant enzyme activities. Histopathological examination further demonstrated that both VpAHPND OMVs and VpTPD OMVs induced varying degrees of structural damage in the hepatopancreas and intestinal tissues. Comparative proteomic analysis of the hepatopancreas provided additional insights into the potential molecular mechanisms underlying shrimp-OMVs interactions. OMVs stimulation triggered coordinated intracellular xenobiotic clearance pathways involving cytoskeleton remodeling-mediated endocytosis-lysosome trafficking, autophagic degradation, and ubiquitin-dependent proteasomal processes, progressed toward programmed cell fate determination characterized by necroptosis, and ultimately drove systemic defensive responses associated with immune recognition and redox homeostasis. Collectively, this study systematically delineates the tissue targeting, cellular responses, and molecular signatures of L. vannamei following stimulation with Vibrio-derived OMVs. These findings suggest that Vibrio OMVs act as critical immunomodulatory signals capable of inducing coordinated systemic and tissue-specific immune responses, providing a foundation for future investigations into their roles in Vibrio-associated diseases and immune regulation in crustaceans.