Characterization of alpha 5-integrin-dependent mast cell adhesion following Fc epsilon RI aggregation.
BACKGROUND
Integrin receptors are engaged in the upregulation of mast cell adhesion to extracellular matrix components upon stimulation with cytokines and antigen. Fibronectin receptor containing the alpha 5-integrin subunit is critical for mast cell interaction with the extracellular matrix protein fibronectin (FN).


METHODS
The murine MCP5/L mast cell line was employed to investigate the process of Fc epsilon RI-mediated mast cell adhesion to FN. RT-PCR and cytofluorimetric analysis were used to assess the expression of alpha 5 integrin in MCP5/L mast cells. Radiolabelled mast cells were sensitized with monoclonal IgE and used in adhesion assays. Anti-alpha 5-integrin antibody (Ab), monovalent hapten and metabolic inhibitors were used to characterize antigen-mediated mast cell adhesion to FN.


RESULTS
Addition of antigen to IgE-sensitized cells resulted in transient upregulation of mast cell adhesion to FN with a maximum adhesion following 30 min of incubation. Mast cell adhesion was inhibited with anti-alpha 5-integrin monoclonal antibodies blocking FN receptor or with excess monovalent hapten preventing antigen-mediated IgE cross-linking. The presence of the protein kinase C (PKC) inhibitor staurosporine also inhibited mast cell adhesion in a dose-dependent fashion. The process of Fc epsilon RI-mediated upregulation of mast cell adhesion to FN was not associated with an increase in surface expression of mast cell FN receptors.


CONCLUSIONS
The major FN receptor on MCP5/L mast cell surface, an integrin containing the alpha 5 subunit mediates a transient change in mast cell adhesiveness following IgE cross-linking. Fc epsilon RI-derived signals engage PKC and upregulate mast cell adhesion in a process which might involve changes in integrin avidity rather than integrin expression.
Dastych
J
J
Department of Biogenic Amines, Polish Academy of Sciences, Lodz, Poland. jdastych@iimcb.gov.pl
Wyczółkowska
J
J
Metcalfe
D D
DD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D015153:Blotting, Western; D002448:Cell Adhesion; D002460:Cell Line; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005353:Fibronectins; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D007152:Immunologic Capping / Q000187:drug effects; D039482:Integrin alpha5; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D017469:Receptors, Fibronectin / Q000737:chemistry / Q000502:physiology*; D017455:Receptors, IgE / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D019311:Staurosporine / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D004791:Enzyme Inhibitors; D005353:Fibronectins; D039482:Integrin alpha5; D017469:Receptors, Fibronectin; D017455:Receptors, IgE; D011493:Protein Kinase C; D019311:Staurosporine