Tissue-resident alveolar macrophages reduce O₃-induced inflammation via MerTK mediated efferocytosis

Lung inflammation, caused by acute exposure to ozone (O₃) - one of the six criteria air pollutants - is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung and their number increases following O₃ exposure. However, the role of AMØs in promoting or limiting O₃-induced lung inflammation has not been clearly defined. Here, we used a mouse model of acute O₃ exposure, lineage tracing, genetic knockouts, and data from O₃-exposed human volunteers to define the role and ontogeny of AMØs during acute O₃ exposure. Lineage tracing experiments showed that 12, 24, and 72 h after exposure to O₃ (2 ppm) for 3h all AMØs were tissue-resident origin. Similarly, in humans exposed to FA and O₃ (200 ppb) for 135 minutes, we did not observe ~21h post-exposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O₃ exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØ demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK - a key receptor involved in efferocytosis - also resulted in impaired clearance of apoptotic neutrophils followed O₃ exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O₃-induced inflammation via MerTK-mediated efferocytosis.