Binding of monoclonal antibody AA4 to gangliosides on rat basophilic leukemia cells produces changes similar to those seen with Fc epsilon receptor activation.
The mAb AA4 binds to novel derivatives of the ganglioside Gd1b on rat basophilic leukemia (RBL-2H3) cells. Some of the gangliosides are located close to the high affinity IgE receptor (Fc epsilon RI), and binding of mAb AA4 inhibits Fc epsilon RI-mediated histamine release. In the present study, mAb AA4 was found to bind exclusively to mast cells in all rat tissues examined. In vitro, within 1 min of mAb AA4 binding, the cells underwent striking morphologic changes. They lost their normal spindle shaped appearance, increased their ruffling, and spread over the surface of the culture dish. These changes were accompanied by a redistribution of the cytoskeletal elements, actin, tubulin, and vimentin, but only the actin was associated with the membrane ruffles. Binding of mAb AA4 also induces a rise in intracellular calcium, stimulates phosphatidyl inositol breakdown, and activates PKC. However, the extent of these changes was less than that observed when the cells were stimulated with antigen or antibody directed against the Fc epsilon RI. None of these changes associated with mAb AA4 binding were seen when the cells were exposed to nonspecific IgG, IgE, or four other anti-cell surface antibodies, nor were the changes induced by binding mAb AA4 at 4 degrees C or in the absence of extracellular calcium. Although mAb AA4 does not stimulate histamine release, it enhances the effect of the calcium ionophore A23187 mediated release. The morphological and biochemical effects produced by mAb AA4 are similar to those seen following activation of the cell through the IgE receptor. Therefore, the surface gangliosides which bind mAb AA4 may function in modulating secretory events.
Laboratory of Immunology, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892.
C
C
Oliver
N
N
Sahara
S
S
Kitani
A R
AR
Robbins
L M
LM
Mertz
R P
RP
Siraganian
D000199:Actins / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000648:ultrastructure; D003599:Cytoskeleton / Q000648:ultrastructure; D005732:Gangliosides / Q000276:immunology / Q000502:physiology*; D006636:Histamine Release; D007073:Immunoglobulin E; D015471:Leukemia, Basophilic, Acute; D008407:Mast Cells / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008870:Microtubules / Q000648:ultrastructure; D011493:Protein Kinase C / Q000378:metabolism; D011961:Receptors, Fc / Q000378:metabolism*; D017455:Receptors, IgE; D014404:Tubulin / Q000032:analysis; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000032:analysis
D000199:Actins; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D005732:Gangliosides; D011961:Receptors, Fc; D017455:Receptors, IgE; D014404:Tubulin; D014746:Vimentin; C033851:ganglioside, GD1b; D007073:Immunoglobulin E; D011493:Protein Kinase C; D002118:Calcium