asthma and airway inflammation
Respiratory assays
Asthma/Airway Inflammation
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Asthma is a chronic inflammatory disorder arising from a poorly understood heterogenic gene-environment interaction. Characteristic features include variable airway obstruction and bronchial hyperresponsiveness. It is frequently known to coexist with atopic diseases, particularly allergic rhinitis. At Cellomatics we can offer a wide range of cellular models and readouts to address the main features of asthma and airway inflammation suitable for both early phase target identification and compound screening.Â
Expression levels of contractile-related genes
Contractile gene expression is altered in HBSMCs following epithelial/immune co-culture.
HBSMCs were cultured alone or co-cultured with BEAS-2B (apical Poly I:C) and/or PBMCs (basolateral LPS) for 48 hours; ACTA2, CNN1, and MYLK were quantified by qPCR (n=3). (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
Cytokine expression from basal chamber
Basal cytokines in BEAS-2B/HBSMC/PBMC co-culture.
HBSMCs ± BEAS-2B (Poly I:C apical) ± PBMCs (LPS basolateral) cultured for 48 hours; basal supernatant Luminex, n=3. (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
Cytokine expression from apical chamber
Apical cytokines in BEAS-2B/HBSMC/PBMC co-culture.
HBSMCs ± BEAS-2B (Poly I:C apical) ± PBMCs (LPS basolateral) cultured for 48 hours; apical supernatant Luminex, n=3. (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
Cytokine expression with TNF-alpha stimulation
Effect of TNF-α and Hydrocortisone on the release of inflammatory mediators by Primary Human Lung Fibroblasts.
Cells were treated with either TNF-α or TNF-α plus Hydrocortisone for 48 hours. The levels of α-SMA, IFN-γ, IL-1β, IL-6, IL-8, HMGB-1, MMP-1, and PAI-1 released in the supernatants was measured by Luminex assay. One-way ANOVA was performed to determine statistical significance (n=3±SEM; *p<0.05;**p<0.01; ***p <0.001).
Cytokine expression with LPS stimulation
LPS increases IL-6 and IL-8 release in primary human lung fibroblasts.
Primary human lung fibroblasts were treated with LPS for 24 hours in the presence or absence of hydrocortisone; IL-6 and IL-8 measured by ELISA (n=5) (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
Airway epithelial-driven allergic airway inflammation
Dexamethasone attenuates IL-1β-driven inflammatory cytokine release in BEAS-2B cells.
BEAS-2B cells were stimulated with IL-1β and treated with Dexamethasone for 48 hours; inflammatory markers in culture supernatants were quantified by Luminex assay (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001; ND=not detected)
Cytokine release with Poly I:C stimulation
PDE4 inhibition reduces Poly I:C–induced IL-6 and IL-8 release in primary human airway smooth muscle cells.
Primary human airway smooth muscle cells were pre-treated with a PDE4 inhibitor and stimulated with Poly I:C for 24 h; IL-6 and IL-8 in supernatants were measured by ELISA (n=3). (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
Cytokine release with Poly I:C stimulation
Poly I:C drives cytokine release from HASMCs and is reduced by TLR3 antagonism.
Human Airway Smooth Muscle Cells (HASMCs) were pretreated with a TLR3 antagonist then stimulated with Poly I:C; supernatants were analysed by Luminex Assay for a panel of cytokines including Eotaxin (n=3). (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05,**p<0.01, ***p<0.001)
Collagen Gel contraction
2,3-BDM reduces collagen gel contraction in airway smooth muscle cells.
Collagen gels with primary human airway smooth muscle cells were pre-treated with 2,3-BDM, stimulated with carbachol for 4 hours, and contraction was quantified as % change vs time 0 (n=3) (Stats: one-way ANOVA; n as stated; mean ± SEM; *p<0.05, **p<0.01, ***p<0.001)
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