Dissecting Immune-Epithelial Interactions in Airway Infection at Single-Cell Resolution Using a Compartmentalised Microfluidic Device

The immune-epithelial interactions that initiate and progress airway diseases have been hard to study using current in vitro models. While Transwell and lung-on-chip systems can mimic airway compartmentalization and allow immune-epithelial contact, they fall short in both spatial resolution and analytical capabilities to track dynamic immune responses during infection.

The Single Cell resolved Airway-Immune Recruitment (scAIR) platform addresses these limitations. It features a modular central chamber with a Transwell insert containing primary airway epithelial cells (AECs) differentiated under air-liquid interface conditions. Immune cells are housed in two flanking compartments connected via a precision microchannel array. This setup enables real-time single-cell imaging of immune cell migration while maintaining epithelial physiology.

By integrating with a machine learning analysis (MLA) pipeline, the scAIR device automates tracking and quantification of individual immune cell movement. This includes measuring speed, direction, and behavioral heterogeneity. Using this platform, researchers can model respiratory syncytial virus (RSV) infection, generating a type 1 inflammatory airway epithelium that promotes neutrophil recruitment.

When TNF-alpha is neutralized with adalimumab, distinct migratory behaviors become apparent, which would be obscured by population-averaged measurements. This integrated platform quantifies airway immune responses during infection and evaluates therapeutic modulation. It allows for mechanistic studies, drug evaluation, and precision modeling of airway inflammation.