Validation of a simplified in vitro Transwell

Area of Science:

• * Pulmonary immunology and host-pathogen interactions.
• * In vitro modeling of respiratory infections.

Background:

• * Aspergillus fumigatus is an opportunistic fungal pathogen that can cause severe lung infections.
• * Understanding host-pathogen interactions at the alveolar level is critical for developing effective treatments.

Purpose of the Study:

• * To develop and validate a simplified in vitro Transwell® model of the human alveolar surface.
• * To investigate the antifungal activity of monocyte-derived dendritic cells (moDCs) against A. fumigatus.
• * To analyze cytokine dynamics and epithelial barrier integrity during fungal invasion.

Main Methods:

• * A Transwell® system with A549 cells modeled the alveolar epithelium.
• * A. fumigatus and moDCs were introduced to simulate lung infection dynamics.
• * Galactomannan assays, LDH measurements, dextran blue permeability, and cytokine profiling were performed.

Main Results:

• * The model accurately simulated fungal invasion and validated by galactomannan assays.
• * moDCs significantly reduced fungal burden, indicating potent antifungal activity.
• * The epithelial barrier remained intact, and cytokine release patterns varied by chamber and molecular weight.

Conclusions:

• * The developed Transwell® model provides a reproducible platform for studying fungal lung infections.
• * moDCs play a crucial role in controlling A. fumigatus growth.
• * The model facilitates dynamic measurement of fungal penetration and host immune responses.