Single-cell proteome profiling reveals distinct immunological patterns in the lungs of ARDS patients

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Rationale: Acute Respiratory Distress Syndrome (ARDS) is a life-threatening condition characterized by lung inflammation and immune dysregulation. Understanding the host immune response is key to identifying ARDS subphenotypes and developing targeted therapies. Methods: Bronchoalveolar lavage (BAL) fluid was repeatedly collected from intubated patients in ICU. BAL leukocytes were analyzed using high-dimensional cytometry by time-of-flight (CyTOF) with a 50-marker panel to assess the maturity, activation and targeted therapeutic potential of immune cells. Results: A total of 128 BAL fluid samples from 91 intubated patients were analyzed. Of these patients, 75(82.4%) had ARDS, 64(70.3%) had pneumonia, and 68(74.7%) survived 28 days post-intubation. Immune cell subsets did not correlate with disease etiology or survival, with immature neutrophils (CD10-) predominant across all disease stages. CD64 expression on neutrophils and PD-1 expression on T cells were higher in pneumonia patients (p<0.001). Macrophage activation markers (pJNK,iNOS,CD163) were negatively associated with mortality, while neutrophil activation markers (CD64,CD11b,CD45RO) decreased in non-survivors. Conclusion: Despite the heterogenous etiology of ARDS, patients share similar immune cell populations in the lungs. However, the activation states of these immune cells vary depending on the presence of pneumonia and are associated with mortality. erj;66/suppl_69/OA1301/F1 F1 F1

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