Biochemical analysis of initial triggering events of IgE-mediated histamine release from human lung mast cells.
Mast cells were isolated from human lung tissues by counter current centrifugation elutriation, followed by flotation through Percoll gradients. Purified human mast cells released histamine upon challenge with anti-IgE. An optimal concentration of anti-IgE for maximum histamine release from human lung mast cells was comparable to that required for histamine release from normal human basophil granulocytes. Human lung mast cells could be passively sensitized with mouse monoclonal IgE antibody for antigen-induced histamine release. Bridging of cell-bound IgE molecules on human mast cells by anti-IgE or its F(ab')2 fragments induced phospholipid methylation and an increase in intracellular cyclic AMP (cAMP). Incorporation of [3H]methyl groups into phospholipid reached a maximum at 30 sec after challenge with anti-IgE, whereas intracellular cAMP reached a maximum at 1 min. Both values declined to base line levels within 2 to 3 min. These biochemical events were followed by Ca2+ influx and histamine release. Ca2+ uptake and histamine release reached maximum at 2 to 3 min and 5 to 8 min, respectively. Neither phospholipid methylation nor initial rise in cAMP was inhibited by indomethacin, which indicates that these biochemical events are not the result of prostaglandin synthesis. However, inhibition of phospholipid methylation by inhibitors of S-adenosyl-L-methionine-mediated methylation, such as 3-deazaadenosine and S-isobutyryl 3-deazaadenosine, inhibited not only phospholipid methylation but also cAMP rise and subsequent Ca2+ uptake and histamine release. The results indicate that phospholipid methylation induced by bridging of IgE receptors on human mast cells is essential for Ca2+ influx and histamine release.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D008168:Lung / Q000166:cytology; D008407:Mast Cells / Q000276:immunology*; D008745:Methylation; D051379:Mice; D010743:Phospholipids / Q000378:metabolism; D011817:Rabbits; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000378:metabolism
D000888:Antibodies, Anti-Idiotypic; D010743:Phospholipids; D017455:Receptors, IgE; D011971:Receptors, Immunologic; C026408:anti-IgE antibodies; D007073:Immunoglobulin E; D000242:Cyclic AMP; D002118:Calcium