Effects of integrin clustering on human lung mast cells and basophils.
The interaction of cells with the extracellular matrix can alter cell responses and is regulated by integrins on the cell surface. We used monoclonal antibodies to the VLA-4 integrins CD29 and CD49d followed by an F(ab')2 fragment of rabbit anti-mouse immunoglobulin G1 to crosslink integrins on the surface of human lung mast cells and basophils. Crosslinking either CD29 or CD49d caused a significant histamine release (HR) from the basophils of most asthmatic donors (10 of 14 for CD49d and 7 of 10 for CD29) (HR = 21 +/- 5%, n = 10, P < 0.005 for CD29 and HR = 19 +/- 4%, n = 14, P < 0.01 for CD49d) yet failed to initiate HR from the basophils of non-atopic and atopic donors (HR was 1 +/- 0.5% for CD29 and 1 +/- 0.5% for CD49d, n = 10, P = NS). Crosslinking either CD29 or CD49d also failed to initiate histamine release from human lung mast cells (HR was 1 +/- 1% for CD29 and 2 +/- 1% for CD49d). The basophils of asthmatic donors responded to 100 and 30 micrograms/ml tissue fibronectin (HR = 12 +/- 2% and 10 +/- 3% for 100 and 30 micrograms/ml fibronectin, respectively, n = 18, P < 0.05), whereas basophils of nonasthmatic patients again failed to degranulate (HR was 0 +/- 0.4% and 1 +/- 0.6%, respectively, n = 11, P = NS). In contrast to the basophil, crosslinking of either CD29 or CD49d failed to initiate histamine release in human lung mast cells (HR = 1 +/- 1% for CD29 and 2 +/- 1%, n = 15). Human lung mast cells were also unresponsive to tissue fibronectin (100 and 30 micrograms/ml) (HR = 1 +/- 1%, n = 5). The tyrosine kinase inhibitor, genistein, significantly reduced CD29- and CD49d-induced HR (inhibition = 83 +/- 7% for CD29 and 77 +/- 6% for CD49d, n > or = 5, P < 0.05). A second tyrosine kinase inhibitor, piceatannol, also significantly reduced both CD29- and CD49d-induced HR (inhibition was 62 +/- 19% for CD29 and 56 +/- 14% for CD49d, n = 7, P < or = 0.05). Integrin crosslinking also affected the response to a second, immunoglobulin E (IgE)-dependent stimulus. Both CD29 and CD49d clustering significantly inhibited anti-IgE-induced histamine release from the human basophil. Inhibition was 30 +/- 5%, n = 18, P < or = 0.001 for CD29 versus 40 +/- 6% for CD49d. In summary, we have shown that crosslinking the beta 1 integrins using either monoclonal antibodies or tissue fibronectin can initiate mediator release from the basophils of asthmatic patients by a mechanism which appears to be tyrosine kinase-mediated. In addition, clustering of integrins modulates the response to a second IgE-dependent signal.
Dept. of Physiology and Pharmacology, University of Southampton, Bassett Crescent East, United Kingdom.
S E
SE
Lavens
K
K
Goldring
L H
LH
Thomas
J A
JA
Warner
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000737:chemistry / Q000276:immunology / Q000502:physiology; D001491:Basophils / Q000502:physiology*; D015550:Cell Degranulation / Q000187:drug effects; D003432:Cross-Linking Reagents; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005353:Fibronectins / Q000494:pharmacology; D019833:Genistein; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology; D039441:Integrin alpha4; D019012:Integrin beta1 / Q000737:chemistry / Q000276:immunology / Q000502:physiology; D016023:Integrins / Q000737:chemistry* / Q000502:physiology*; D007529:Isoflavones / Q000494:pharmacology; D008168:Lung / Q000166:cytology*; D008407:Mast Cells / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D013267:Stilbenes / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D003432:Cross-Linking Reagents; D004791:Enzyme Inhibitors; D005353:Fibronectins; D019012:Integrin beta1; D016023:Integrins; D007529:Isoflavones; D013267:Stilbenes; D039441:Integrin alpha4; D007073:Immunoglobulin E; C041525:3,3',4,5'-tetrahydroxystilbene; D019833:Genistein; D011505:Protein-Tyrosine Kinases