Cross-Linking Mast Cell Specific Gangliosides Stimulates the Release of Newly Formed Lipid Mediators and Newly Synthesized Cytokines
Cross-linking GD1b derived gangliosides by mAbAA4 induced Syk-dependent release of PGD2 and PGE2. For stimulation via FcεRI, RBL-2H3 cells and C4A2 Syk-negative cells were sensitized with IgE anti-TNP and stimulated with DNP48-HSA (50 ng/mL). For FcεRI independent stimulation, the cells were incubated with calcium ionophore (0.1 μg/mL). To cross-link GD1b derived gangliosides, cells were incubated with mAbAA4 (1, 2.5, 5, and 10 μg/mL). Nonstimulated (NS) cells were used as negative controls. Culture supernatants were collected after 30 min of incubation to evaluate immediate release (a and c) or cells were incubated for 1 h and rinsed and cultured for an additional 3 h and culture supernatants were collected to evaluate delayed release (b and d). PGD2 (a and b) and PGE2 (c and d) were measured in the culture supernatant by EIA. Data is expressed as the mean ± SD of three independent experiments. between experimental samples and the nonstimulated (NS) cells. between experimental samples and FcεRI stimulated RBL-2H3 cells.
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