. 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 (FCcRI), and binding of mAb AA4 inhibits FCERI-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 FcERI. 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°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 .phate by mobilizing intracellular calcium pools . Receptor activation also results in calcium influx, activation of phospholipase A2, the phosphorylation of a number of intracellular proteins (7), and the rearrangement of the cytoskeleton before mediator release (35,37,48) .The rat basophilic leukemia cell line, RBL-2113, has been widely used as a model to study 48,49) . A number ofmAbs have been raised against surface molecules on these cells (3,5,29,50) . These mAb were selected for their ability to either stimulate or inhibit histamine release in the RBL-2113 cells. The majority of the antibodies were directed against the Fcc-RI ; however, two antibodies, mAb ADl and AA4, bind other cell surface components . The mAb ADl binds to a novel 50-60-kD protein on the surface of the RBL-2113 cells and modulates histamine release (31). The other antibody, mAb AA4, binds to unique a-galactosyl derivatives of the ganglioside GD, b (18). These gangliosides are closely related to the FceRl, both spatially and functionally (5) . Binding ofboth the intact mAb AA4 and its Fab fragments to cells inhibits the binding

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Background: Recent data suggest that normal tissue mast cells can express functional receptors for IgG under certain conditions. However, little is known about IgG receptor expression and functional consequences in mast cell neoplasms. Methods: In this study, neoplastic mast cells were obtained from a dog with cutaneous mastocytoma (CM-MC) and from a dog with visceral mastocytoma (VI-MC). Both cell populations were characterized morphologically and functionally. Results: Most cells proliferated constantly in suspension without particular supplements. Doubling times of CM-MC and VI-MC were 52.2 and 27.5 h, respectively. Both cell types were sensitive to formalin fixation, did not contain heparin and were tryptase and chymase positive. Electron microscopy showed fine granules with electron-dense content in both cell populations. The total histamine content of CM-MC and VI-MC was 0.25 and 0.10 pg/cell, respectively. Calcium ionophore A23187 and substance P induced dose-dependent histamine release, whereas compound 48/80 had no effect. Most significantly, both cell types, when sensitized with monomeric dog IgG, released histamine upon stimulation by anti-dog IgG. Conclusions: Dog mastocytoma-derived cells may be useful to study the regulation of neoplastic mast cell growth and differentiation, as well as IgG receptor-mediated activation in neoplastic mast cells. Further research is required to clarify the pathophysiological significance of constitutive expression of IgG receptors in neoplastic (canine) mast cells.

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