Protocol to desensitize human and murine mast cells after polyclonal IgE sensitization

Allergic inflammation triggers dyslipidemia via IgG signalling

“…Then, 700 μL of heat-treated sera was transferred to naive recipients the previous day to challenge. The irreversible denaturation and deactivation of IgE was confirmed by ELISA and mast cell activation test 32 . In the rest of the groups (naive, IgE, IgE+IgG), the naive or native allergic sera, respectively, were serially transferred as follows: 100 µL; 200 µL; and 400 µL with an interval of 48 hours between each administration.…”

“…S6a ). To ascertain if the remnants of denatured IgE were functional, we used a cell activation assay based on bone marrow-derived mast cells 32 . Mast cells sensitized with native allergic sera showed a dose-dependent activation following allergen stimulation by means of CD63 expression detected by flow cytometry ( Fig.…”

Anaphylaxis and desensitization differently activate and induce IL‐8 release by mast cells in a human peanut allergy in vitro model

“…Restimulated desensitized MCs showed little increase in CD63 expression, compared to non-restimulated (7.21 ± 1.87%, p = 0.0048), however still significantly below both anaphylaxis and positive αIgE activation control (Figure 1B), and not followed by significantly increased tryptase release (4.69 ± 5.329, p = 0.2723, Figure S4B). Twenty-four hours from initial activation, leftover expression of CD63 could be observed in non-restimulated anaphylaxis/desensitized MCs due to incomplete membrane recycling, 3 although no further degranulation could be elicited upon restimulation in both anaphylaxis and desensitized MCs (p > 0.05, Figure 1C). MCs were completely refractory for at least 48 h, regardless of the initial stimulation modality.…”

“…Correct timing and dose are fundamental to desensitize MCs, as described using mouse models, 2,3 human cell lines, 3,4 and tissuederived MCs. 5,6 Nevertheless, the differences in activation between anaphylaxis and desensitization in human MCs remain to be elucidated, particularly late-phase activities (cytokine release, degranulation recovery kinetics), and when using human sera as source of allergen-specific IgE.…”

Meglumine gadoterate induces immunoglobulin‐independent human mast cell activation via MRGPRX2

“…with several commercial GBCA, namely, meglumine gadoterate, gadobutrol, gadoxetate disodium, and gadoteridol. Then, we determined cell viability and degranulation by flow cytometry 4 (see a detailed material and methods section in this article's Appendix S1).…”