Gentle Isolation of Eosinophils To Study Lung Inflammation

Gentle Isolation of Eosinophils To Study Lung Inflammation Immune responses to pathogenic threats are tightly regulated, both locally at the tissue level and systemically through the coordinated recruitment of immune cells. Eosinophils are central effectors in allergic airway inflammation, responding to temporal and spatial cues to mount appropriate responses when recruited to the lung. Despite advances from single-cell RNA sequencing, high-parameter flow cytometry, and functional assays, eosinophil plasticity remains poorly understood due to their low transcriptional activity, limited mRNA content, and vulnerability to loss during conventional isolation. Using eosinophil-lineage tracing mice, we show that eosinophil phenotype is strongly shaped by the local lung environment. Recruited eosinophils respond to pro-inflammatory signals like TNF but are restrained by intrinsic negative feedback to prevent excessive inflammation. Importantly, the MACSQuant Tyto Lux Cell Sorter enabled isolation of untouched, highly viable eosinophils directly from inflamed airways, which was previously unachievable with conventional high-pressure sorters. Its gentle, closed-system microfluidics and rapid throughput allowed adoptive transfer and systematic functional interrogation of eosinophil-intrinsic feedback signalling and cytokine production. Eosinophils are central effectors in allergic airway inflammation, responding to temporal and spatial cues to mount appropriate responses when recruited to the lung. Understanding their behavior is critical, yet their fragility and low transcriptional activity have made them challenging to study. Traditional methods often compromise viability, limiting insights into how these cells respond to the lung microenvironment. This webinar will reveal new research uncovering intrinsic regulatory mechanisms of eosinophils and highlight an innovative approach that preserves their function during isolation. By attending, you will gain valuable knowledge on how these advances open the door to deeper exploration of complex immune responses.