Purpose
Tween‐80 is one of the most important causes resulting in anaphylactoid reaction. However, its mechanism remains unclear. Proteomic characterizations of mast cells’ excreta in response to Tween‐80 are assayed to investigate the mechanism of anaphylactoid reaction.
Experimental design
A label‐free LCMS/MS‐based proteomics is used to analyze Tween‐80‐stimulated Laboratory of Allergic Diseases 2 (LAD2) mast cells releasates. The results of proteomic are analyzed by bioinformatics analysis. Western blotting is used to verify the expression of proteins.
Results
Overall, endocytosis, nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), and calcium signaling pathways play important roles in Tween‐80‐induced LAD2 cells activation by bioinformatics analysis. The expressions of relative proteins including actin‐related protein 2/3 complexes, vacuolar protein sorting‐associated protein, phosphorylation of transcription factor of P105 and P65, phosphorylation of inositol 1,4,5‐trisphosphate receptor (IP3R), phosphoinositide phospholipase Cγ (PLCγ), and protein kinase C (PKC), are significantly increased in Tween‐80 group compared to control. Tween‐80 might be internalized via endocytosis, which induces degranulation by PLCγ/PKC pathways mediated calcium influx, and promotes the generation of inflammatory mediators via NF‐κB pathway resulting in anaphylactoid reaction.