Conclusion
Activation through high affinity IgE leads to well-characterised cytokine release. When activating primary human mast cell lines through low affinity IgE with the native allergen Der p 2, some chemokines, and cytoplasmic proteins were released, and surface receptors were solubilized. The impact of finely tuned IgE affinity for allergen to the nature of the mast cell response in both cytokine and chemokine production as well as surface marker profile may modulate the severity of the late phase response and determine the chronic response to repeated allergic activation.
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Figure legends
Figure 1
a: Mast cell activation as CD63 expression after stimulation with recombinant Der p 3 allergen for 30 min at 37C at high (red) and low (blue) IgE affinity.
b: Prostaglandin D2 synthesis from 3 independent cell lines at high (red) and low (blue) IgE affinity. (c-f) Small graphs chart the metabolism of PGD2 through PGJ2 and delta-12-PGJ2 to 15-deoxy-delta-12,14-PGJ2 detected by MS of one cell line.
Figure 2:
Plots of cytokines and chemokines and secretory proteins up-regulated significantly by high-affinity (red) compared to low-affinity (blue) activation, or, low-affinity compared to high-affinity activation after activation. The upregulation is shown as fold change compared with resting (0 hour) mast cells,. Ordinates are dimensioned to illustrate the particular reactants variation. An * next to the reactant indicates significance in the limma analysis, a H andL indicate that a marker is upregulated significantly with high or low affinity IgE. Detailed information is given in table 1.
Figure 3:
Plots of shedded surface markers up-regulated significantly by high-affinity (red) compared to low-affinity (blue) activation, or, low-affinity compared to high-affinity activation after activation. The upregulation is shown as fold change compared with resting (0 hour) mast cells,. Ordinates are dimensioned to illustrate the particular reactants variation. An * next to the reactant indicates significance in the limma analysis, a H and L indicate that a marker is upregulated significantly with high or low affinity IgE. Detailed information is given in table 1.
Figure 4
Secretory proteins detected at different timepoints after activation. The up-regulation is shown as the fold change compared to corresponding resting mast cells (0 hour).
Figure 5
An alternate visualization of the detection of reactants in mast cell supernatants after stimulation through either high or low affinity IgE. The up-regulation is shown as the fold change compared to corresponding resting mast cells (0 hour). At 24 hours the range of upregulation is only 8-fold, where it is 64-fold at 3 and 6 hours.
Table Legends
Table 1
Mean Olink NPX data for all measured reagents at all time points.
Table 2
Results of a limma analysis of changes over time of mast cells activated through either high- or low affinity IgE.