Cross-Linking Mast Cell Specific Gangliosides Stimulates the Release of Newly Formed Lipid Mediators and Newly Synthesized Cytokines

Figure 3

Cross-linking GD1b derived gangliosides with mAbAA4 induced the release of IL-4, IL-6, and TNF-α in a Syk-dependent manner. 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 1 h after stimulation and the cells were rinsed and cultured for an additional 11 h to evaluate cytokine release. IL-4 (a), IL-6 (b), and TNF-α (c) were measured in the culture supernatants by ELISA. 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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