Fibroblast-myofibroblast transition is differentially regulated by bronchial epithelial cells from asthmatic children

Respiratory Research

Jason S Debley¹  Sarah Herrington-Shaner²  Tin-Yu Lien²  Tessa Kolstad²  Stephen R Reeves¹ 

[1] Department of Pediatrics, University of Washington School of Medicine, Seattle, USA;Center for Immunity and Immunotherapies, Jack R. MacDonald Building, Seattle Children’s Research Institute, 1900 9th Ave, Seattle 98109, WA, USA

关键词: TGFβ2;    α-smooth muscle actin;    Myofibroblasts;    Fibroblasts;    Cell culture;    Bronchial epithelial cells;    Asthma;    Airway remodeling;    Air-liquid interface culture;   

Others  :  1133351
DOI  :  10.1186/s12931-015-0185-7

 received in 2014-07-24, accepted in 2015-01-29,  发布年份 2015

【 摘 要 】

Background

Airway remodeling is a proposed mechanism that underlies the persistent loss of lung function associated with childhood asthma. Previous studies have demonstrated that human lung fibroblasts (HLFs) co-cultured with primary human bronchial epithelial cells (BECs) from asthmatic children exhibit greater expression of extracellular matrix (ECM) components compared to co-culture with BECs derived from healthy children. Myofibroblasts represent a population of differentiated fibroblasts that have greater synthetic activity. We hypothesized co-culture with asthmatic BECs would lead to greater fibroblast to myofibroblast transition (FMT) compared to co-culture with healthy BECs.

Methods

BECs were obtained from well-characterized asthmatic and healthy children and were proliferated and differentiated at an air-liquid interface (ALI). BEC-ALI cultures were co-cultured with HLFs for 96 hours. RT-PCR was performed in HLFs for alpha smooth muscle actin (α-SMA) and flow cytometry was used to assay for α-SMA antibody labeling of HLFs. RT-PCR was also preformed for the expression of tropomyosin-I as an additional marker of myofibroblast phenotype. In separate experiments, we investigated the role of TGFβ₂ in BEC-HLF co-cultures using monoclonal antibody inhibition.

Results

Expression of α-SMA by HLFs alone was greater than by HLFs co-cultured with healthy BECs, but not different than α-SMA expression by HLFs co-cultured with asthmatic BECs. Flow cytometry also revealed significantly less α-SMA expression by healthy co-co-cultures compared to asthmatic co-cultures or HLF alone. Monoclonal antibody inhibition of TGFβ₂ led to similar expression of α-SMA between healthy and asthmatic BEC-HLF co-cultures. Expression of topomyosin-I was also significantly increased in HLF co-cultured with asthmatic BECs compared to healthy BEC-HLF co-cultures or HLF cultured alone.

Conclusion

These findings suggest dysregulation of FMT in HLF co-cultured with asthmatic as compared to healthy BECs. Our results suggest TGFβ₂ may be involved in the differential regulation of FMT by asthmatic BECs. These findings further illustrate the importance of BEC-HLF cross-talk in asthmatic airway remodeling.

【 授权许可】

   
2015 Reeves et al.; licensee BioMed Central.

【 图 表 】

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