Tight regulation of glucose concentration in the air surface liquid (ASL) is important for protection against infection. We have previously shown a mechanism for glucose regulation that involves glucose uptake into airway epithelial cells through facilitated glucose transporters, GLUTs (Kalsi et al, 2008). Airway inflammation could disrupt glucose homeostasis in airway epithelial cells via this mechanism. The aim of this study was to investigate the effect of inflammation induced by pro-inflammatory cytokines on GLUT function and expression in these cells. Human H441 and primary human bronchial epithelial cell (HBEC) cultures were used. Cells were grown at air liquid interface for 7-14 days. Inflammation was induced with a cocktail of cytokines [Tumor Necrosis Factor -α (100 ng/ml), Interferon -γ (100 ng/ml), Lipopolysaccharide (10 μg/ml)] for 0, 24, 48 and 72 hours. The monolayer resistance was measured using Ussing chamber and calculated using V=IR. The effect of cytokine treatment on glucose uptake was assessed using radiolabelled glucose (14C-D-glucose, 1μCi) applied to the apical or basolateral surface. GLUT-specific transport was assessed using the pharmacological inhibitor, Phloretin. Western blotting analysis and immunocytochemistry were used to determine GLUT protein expression. Cytokine induced inflammation significantly reduced H441 resistance to 56 ± 4.11%, 32 ± 1.03%, 17 ± 2.09% at 24, 48 & 72 h, respectively of control (P≤0.05, n=3). Preliminary data shows that cytokine treatment caused a similar reduction in resistance of HBECs. The amount of glucose uptake was lower across the apical membrane than the basolateral (P≤0.05, n=3). The phloretin sensitive component of apical glucose uptake in H441 compared to its relative control was 2.04 ± 2.7, 2.96 ± 1.5, 7.15 ± 1.7, 3.67 ± 2.5 nmol glucose/ mg protein with exposure to cytokine treatment at 0, 24, 48 & 72 h, respectively and significance was observed at 48 h (P≤0.05, n=3). However, the basolateral phloretin sensitive component in H441 was not changed (8.06 ± 12.4, 10.89 ± 5.6, 5.65 ± 7.5, 6.51 ± 4.1 nmol glucose/mg protein) with the same duration of cytokine treatment (n=3). GLUT2 and GLUT10 were observed in airways cells by fluorescence immunocytochemistry, the latter was highly expressed in these cells. H441 (total protein) expressed GLUT2, GLUT10 and very low or no expression of GLUT1. The abundance of GLUT2 was not affected by cytokine treatment (n=3). In conclusion, inflammation induced by a cocktail of cytokines had a profound affect on airway epithelial resistance and apical glucose uptake at 48 h. As there were no changes in GLUT2 abundance we speculate that changes in GLUT10 expression may play a role in glucose uptake during induced inflammation.