title: Evidence for a novel Kit adhesion domain mediating human mast cell adhesion to structural airway cells.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire, LE12 5RD, UK. Kevin.gough@nottingham.ac.uk.
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Gough
authors.affiliation: ADAS UK, School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire, LE12 5RD, UK. Benmaddison@addas.co.uk.
authors.forename: Ben C
authors.initials: BC
authors.lastname: Maddison
authors.affiliation: Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, UK. Ap342@le.ac.uk.
authors.forename: Aarti
authors.initials: A
authors.lastname: Shikotra
authors.affiliation: Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, UK. Elena.moiseeva@sky.com.
authors.forename: Elena P
authors.initials: EP
authors.lastname: Moiseeva
authors.affiliation: Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, UK. weidongyang@yahoo.com.
authors.forename: Weidong
authors.initials: W
authors.lastname: Yang
authors.affiliation: ADAS UK, Biology Department, University of Leicester, University Road, Leicester, LE1 7RH, UK. shilajarvis@outlook.com.
authors.forename: Shila
authors.initials: S
authors.lastname: Jarvis
authors.affiliation: Department of Infection, Immunity and Inflammation, Institute for Lung Health, University of Leicester, Leicester, UK. pb46@le.ac.uk.|Department of Respiratory Medicine, Glenfield Hospital, Groby Rd, Leicester, LE3 9QP, UK. pb46@le.ac.uk.
authors.forename: Peter
authors.initials: P
authors.lastname: Bradding
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008407:Mast Cells / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D020545:Respiratory Mucosa / Q000378:metabolism*
chemical_list: D019009:Proto-Oncogene Proteins c-kit