title: Immunologic and ionophore-induced generation of leukotriene B4 from mouse bone marrow-derived mast cells.
abstract: Mouse bone marrow-derived mast cells differentiated in vitro and sensitized with monoclonal IgE respond to antigen-initiated activation-secretion with the generation of leukotriene B4 (LTB4), a highly potent chemotactic factor. The antigen-initiated net percent release of the secretory granule marker beta-hexosaminidase and the generation of immunoreactive LTB4 and of immunoreactive leukotriene C4 (LTC4) were not diminished by washing the cells before challenge, indicating that interaction of the antigen occurred with the IgE fixed on cell membranes and was not due to phagocytosis of immune complexes formed in the fluid phase. The parallel dose-response relationship for the antigen-induced release of the performed mediator and the generation of both leukotrienes along with the superimposable time courses of their extracellular appearance indicate the origin of these mediators from a common cell type with IgE receptors. Resolution by reverse phase-high performance liquid chromatography (RP-HPLC) of the leukotrienes released from unsensitized cells by ionophore A23187 and from sensitized cells by the specific antigen revealed that the generation of LTB4 was accompanied by the production of the two diastereoisomers of 6-trans-LTB4, which were not immunoreactive. The immunoreactive LTB4 eluted from RP-HPLC at the same retention time as synthetic LTB4 was present in similar nanogram quantities when measured by either radioimmunoassay or integrated absorbance at 269 nm and exhibited a chemotactic activity for human neutrophils on a weight basis comparable to that of synthetic LTB4. The overall recoveries after RP-HPLC of immunoreactive LTB4 released from ionophore A23187-activated, bone marrow-derived mast cells and of added tritiated synthetic LTB4 in separate experiments with ionophore-activated cells were comparable and greater than 78%. The maximum generation of LTB4 by ionophore A23187-activated cells of 7.9 +/- 2.5 ng/10(6) cells (mean +/- SD), occurring at 40 min, was greater than the maximum generation of 4.5 +/- 0.8 ng/10(6) cells, with immunologic stimulation occurring 5 min after antigen challenge of sensitized cells. The initial rate of LTB4 generation after perturbation of the IgE-Fc receptors, however, exceeded that following ionophore A23187 stimulation and may represent one important variable in establishing a significant chemotactic gradient.
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001854:Bone Marrow Cells; D000001:Calcimycin / Q000494:pharmacology*; D002417:Cattle; D004136:Dinitrobenzenes / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D007975:Leukotriene B4 / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D008297:Male; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D012710:Serum Albumin, Bovine / Q000276:immunology
chemical_list: D000900:Anti-Bacterial Agents; D004136:Dinitrobenzenes; D007975:Leukotriene B4; D012710:Serum Albumin, Bovine; D007073:Immunoglobulin E; D000001:Calcimycin