Particulate matter (PM) with a median diameter <2.5 μm, is associated with respiratory and cardiovascular diseases. We previously reported the biological effects of PM in vivo, and although neutrophils play an important role in initiating inflammation, few reports have focused on the relationship between PM inhalation and immune responses. Here, we investigated the effect of PM particle size on neutrophils, including their endocytosis activity and reactive oxygen species (ROS) production. Flow cytometry analysis indicated that 1 μm particles are readily endocytosed by neutrophils and that endocytosis is reduced at 4 °C. Inhibitors of the pleckstrin homology domain of dynamin repressed this process; however, GTPase and clathrin inhibitors did not affect endocytosis. Endocytosis by neutrophils in Toll-like receptor 4 (TLR4)- and MyD88-knockout mice was reduced compared with that in wild-type mice, indicating that TLR4 and MyD88 are important for the process. Neutrophil-mediated endocytosis caused oxidative stress, and N-acetylcysteine enhanced endocytosis. Expression levels of the oxidative stress markers, heme oxygenase-1 and p62 protein, were increased in an endocytosis-dependent manner. Phagocytosed neutrophils produced IL-6 and TNFα, whose production was decreased by dynamin inhibitors. We observed that infiltrated CD11b-positive cells in bronchoalveolar lavage fluid endocytose PMs. Overall, these results indicate that endocytosis and ROS production via TLR4 are important for the initiation of immune responses by neutrophils.