Evidence for a novel Kit adhesion domain mediating human mast cell adhesion to structural airway cells

Respiratory Research

Peter Bradding⁵  Shila Jarvis¹  Weidong Yang²  Elena P. Moiseeva²  Aarti Shikotra²  Ben C. Maddison³  Kevin C. Gough⁴ 

[1] ADAS UK, Biology Department, University of Leicester, University Road, Leicester LE1 7RH, UK;Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, UK;ADAS UK, School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire LE12 5RD, UK;School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire LE12 5RD, UK;Department of Respiratory Medicine, Glenfield Hospital, Groby Rd, Leicester LE3 9QP, UK

关键词: Kit (CD117);    ScFv;    Phage display;    Airway smooth muscle;    Airway epithelium;    Mast cell;   

Others  :  1227515
DOI  :  10.1186/s12931-015-0245-z

 received in 2014-10-30, accepted in 2015-07-01,  发布年份 2015

【 摘 要 】

Background

Human lung mast cells (HLMCs) infiltrate the airway epithelium and airway smooth muscle (ASM) in asthmatic airways. The mechanism of HLMC adhesion to both cell types is only partly defined, and adhesion is not inhibited by function-blocking anti-Kit and anti-stem cell factor (SCF) antibodies. Our aim was to identify adhesion molecules expressed by human mast cells that mediate adhesion to human ASM cells (HASMCs) and human airway epithelial cells.

Methods

We used phage-display to isolate single chain Fv (scFv) antibodies with adhesion-blocking properties from rabbits immunised with HLMC and HMC-1 membrane proteins.

Results

Post-immune rabbit serum labelled HLMCs in flow cytometry and inhibited their adhesion to human BEAS-2B epithelial cells. Mast cell-specific scFvs were identified which labelled mast cells but not Jurkat cells by flow cytometry. Of these, one scFv (A1) consistently inhibited mast cell adhesion to HASMCs and BEAS-2B epithelial cells by about 30 %. A1 immunoprecipitated Kit (CD117) from HMC-1 lysates and bound to a human Kit-expressing mouse mast cell line, but did not interfere with SCF-dependent Kit signalling.

Conclusion

Kit contributes to human mast cell adhesion to human airway epithelial cells and HASMCs, but may utilise a previously unidentified adhesion domain that lies outside the SCF binding site. Targeting this adhesion pathway might offer a novel approach for the inhibition of mast cell interactions with structural airway cells, without detrimental effects on Kit signalling in other tissues.

【 授权许可】

   
2015 Gough et al.

【 图 表 】

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