Vesicle associated membrane protein (VAMP)-7 and VAMP-8, but not VAMP-2 or VAMP-3, are required for activation-induced degranulation of mature human mast cells

• Published in: European journal of immunology 2008-03, Vol.38 (3), p.855-863
• Main Authors: Sander, Leif E, Frank, Simon P.C, Bolat, Seza, Blank, Ulrich, Galli, Thierry, Bigalke, Hans, Bischoff, Stephan C, Lorentz, Axel
• Format: Article
• Language: English
• Subjects: Allergology; Antibodies; Antibodies - pharmacology; Blotting, Western; Cell Degranulation; Cell Degranulation - drug effects; Cell Degranulation - physiology; Cell Membrane; Cell Membrane - metabolism; Cellular immunology; Cytoplasm; Cytoplasm - metabolism; Fluorescent Antibody Technique; Gene Expression; Gene Expression - drug effects; Histamine Release; Histamine Release - drug effects; Histamine Release - physiology; Humans; Immunology; Life Sciences; Mast Cells; Mast Cells - drug effects; Mast Cells - metabolism; Mast Cells - physiology; Metalloendopeptidases; Metalloendopeptidases - pharmacology; Mucosal immunity; Qa-SNARE Proteins; Qa-SNARE Proteins - genetics; Qa-SNARE Proteins - immunology; Qa-SNARE Proteins - metabolism; Qb-SNARE Proteins; Qb-SNARE Proteins - genetics; Qb-SNARE Proteins - immunology; Qb-SNARE Proteins - metabolism; Qc-SNARE Proteins; Qc-SNARE Proteins - genetics; Qc-SNARE Proteins - immunology; Qc-SNARE Proteins - metabolism; R-SNARE Proteins; R-SNARE Proteins - genetics; R-SNARE Proteins - metabolism; R-SNARE Proteins - physiology; Reverse Transcriptase Polymerase Chain Reaction; SNARE Proteins; SNARE Proteins - genetics; SNARE Proteins - metabolism; Tetanus Toxin; Tetanus Toxin - pharmacology; Vesicle-Associated Membrane Protein 2; Vesicle-Associated Membrane Protein 2 - genetics; Vesicle-Associated Membrane Protein 2 - metabolism; Vesicle-Associated Membrane Protein 2 - physiology; Vesicle-Associated Membrane Protein 3; Vesicle-Associated Membrane Protein 3 - genetics; Vesicle-Associated Membrane Protein 3 - metabolism; Vesicle-Associated Membrane Protein 3 - physiology
• ISSN: 0014-2980; 1521-4141
• DOI: 10.1002/eji.200737634

Mediator release from mast cells (MC) is a crucial step in allergic and non-allergic inflammatory disorders. However, the final events in response to activation leading to membrane fusion and thereby facilitating degranulation have hitherto not been analyzed in human MC. Soluble N-ethyl-maleimide-sensitive factor attachment protein receptors (SNARE) represent a highly conserved family of proteins that have been shown to mediate intracellular membrane fusion events. Here, we show that mature MC isolated from human intestinal tissue express soluble N-ethylmaleide sensitive factor attachment protein (SNAP)-23, Syntaxin (STX)-1B, STX-2, STX-3, STX-4, and STX-6 but not SNAP-25. Furthermore, we found that primary human MC express substantial amounts of vesicle associated membrane protein (VAMP)-3, VAMP-7 and VAMP-8 and, in contrast to previous reports about rodent MC, only low levels of VAMP-2. Furthermore, VAMP-7 and VAMP-8 were found to translocate to the plasma membrane and interact with SNAP-23 and STX-4 upon activation. Inhibition of SNAP-23, STX-4, VAMP-7 or VAMP-8, but not VAMP-2 or VAMP-3, resulted in a markedly reduced high-affinity IgE receptor-mediated histamine release. In summary, our data show that mature human MC express a specific pattern of SNARE and that VAMP-7 and VAMP-8, but not VAMP-2, are required for rapid degranulation.