An IgE-dependent secretory response of mast cells can be induced by a glycosphingolipid-specific monoclonal antibody.
The signal transduction pathway of the type 1 Fcepsilon receptor (FcepsilonRI) has been proposed to be spatially constrained to plasma membrane microdomains enriched in glycosphingolipids and cholesterol. These domains are proposed to serve as platforms that enhance the efficiency of the antigen-receptor stimulus-response coupling process. Here we describe a monoclonal antibody (mAb) designated 2B5, raised by immunizing mice with rat mucosal-type mast cell (line RBL-2H3) membranes, which binds to glycosphingolipids and causes a dose-dependent secretory response of these cells. This secretory response to mAb 2B5 requires binding of IgE to the FcepsilonRI on these cells, although direct interactions between IgE and mAb 2B5 are excluded. The bound IgE- or FcepsilonRI-specific mAb did not affect binding of mAb 2B5 or its Fab fragments to the RBL-2H3 cells and only a limited interference with the binding of IgE to the FcepsilonRI by mAb 2B5 was observed. Binding of mAb 2B5 to the RBL-2H3 cells induced a distribution of fluorescently labeled IgE similar to that produced by antigen-induced aggregation of the IgE-FcepsilonRI. Thus we suggest that mAb 2B5 binds to cell surface glycosphingolipids that are probably associated with the FcepsilonRI-IgE complexes and causes their aggregation, thereby initiating the cascade leading to the cell's secretory response.
Schwarz
A
A
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
Jürgens
L
L
Licht
A
A
Schneider
H
H
Futerman
A H
AH
Pecht
I
I
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D004306:Dose-Response Relationship, Immunologic; D006028:Glycosphingolipids / Q000502:physiology*; D007073:Immunoglobulin E / Q000502:physiology*; D008407:Mast Cells / Q000378:metabolism*; D051381:Rats; D017455:Receptors, IgE / Q000502:physiology
D000911:Antibodies, Monoclonal; D006028:Glycosphingolipids; D017455:Receptors, IgE; D007073:Immunoglobulin E