Ex vivo air–liquid interface respiratory mucosa platform to study type 2–driven Asthma

: Asthma is a heterogeneous disorder in which a subset of patients exhibits a type 2 (Th2- or T2-high) endotype driven by eosinophilic, IL-4/IL-13-mediated inflammation. Traditional animal and 2D cell-based models incompletely reproduce human airway immune responses, particularly the Th2 phenotype. To address this gap, we developed a three-dimensional (3D) ex vivo model of human nasal respiratory mucosa incorporating type-2-biased immune stimulation. Primary nasal mucosal biopsies were expanded under air-liquid interface (ALI) conditions and either exposed to IL-4/IL-13 or co-cultured with autologous polarized CD4+ Th2 lymphocytes and dendritic cells to generate a Th2-dominated microenvironment. Tissue morphology and barrier function were monitored longitudinally by phase-contrast microscopy and transepithelial electrical resistance (TEER). Induction of a type-2 inflammatory state was confirmed by increased secretion of periostin, STAT-6, IL-4 and IL-13 in apical and basal compartments, together with modest TEER reduction and evidence of epithelial remodeling, whereas IL-8 and chitinase family proteins did not increase, thereby excluding a COPD-like or type-2-low phenotype. Rather than fully recapitulating clinical asthma, this methodological model reproduces key immunopathologic features of T2-high airway inflammation in a patient-derived 3D context. It provides a stable, human-relevant platform for mechanistic studies and for future preclinical screening of targeted anti-inflammatory therapies.