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Updated: Sep 2, 2025

An Ex vivo Mast Cell Degranulation Assay using Crude Peritoneal Exudate Cells and Natural Antigen Stimulation
Published on: April 27, 2021
Specific histamine regulating activity of surface-modified yeast vacuoles by histamine- binding protein and its
Hyeweon Jang1, Yang-Hoon Kim2, Jiho Min1
1Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University, Jeonju-si, Korea.
Abstract:
We aimed to develop a biocompatible material that could enhance weakened immunity and control histamine in vivo. Histamine-binding protein (HBP) vacuoles have a mechanism of action that directly binds to the histamine molecule. It is designed to eliminate the side effects of antihistamine caused by binding to other receptors. HBP vacuoles were designed to produce a material that was biocompatible, and could enhance immunity. First, a recombinant vector was designed so that HBP was located on the vacuole surface, and expressed towards the cytoplasm. The vector was transformed into yeast, and protein expression was induced. Then, the vacuole was isolated by centrifugation to complete HBP vacuoles. Cytotoxicity test was conducted for application to RAW 264.7 cells. In addition, immune enhancement reaction and histamine inhibition were confirmed through phagocytosis assay and histamine ELISA. RAW 264.7 cells were pre-treated with HBP vacuoles to confirm the immune enhancement of HBP vacuoles. As a result, it was confirmed that the immunostimulatory effect of the vacuole was increased in a concentration-dependent manner. In addition, the reduction of histamine was confirmed by treating the HBP vacuoles. As a result, HBP vacuoles reduced the histamine secreted from RAW 264.7 cells by about 75%.
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