Real-time detection of mast cell degranulation in anaphylactoid reaction.
- Author:
Jianjiang HU
1
;
Yanming HOU
;
Qian ZHANG
;
Hongtao LEI
;
Yi WANG
;
Danqiao WANG
Author Information
- Publication Type:Journal Article
- MeSH: Anaphylaxis; diagnosis; immunology; metabolism; Animals; Antigens, CD; genetics; Cell Degranulation; Cell Line, Tumor; Cell Movement; Mast Cells; cytology; immunology; Microscopy, Confocal; Microscopy, Fluorescence; Platelet Membrane Glycoproteins; genetics; Rats; Tetraspanin 30; Time Factors
- From: China Journal of Chinese Materia Medica 2011;36(14):1860-1864
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a new, real time, dynamic and direct optical detection method for mast cell degranulation caused by anaphylactoid reaction.
METHODA CD63-GFP plasmid was constructed and introduced steadily into rat basophilic leukemia (RBL-2H3) cells. The movements of CD63-GFP, which was located on both the granule membranes and the plasma membranes of RBL cells stimulated by Compound 48/80, were studied by confocal laser scanning microscope (CLSM) and total internal reflection fluorescence microscope (TIRFM) both inside and on the surface of living RBL-2H3 cells.
RESULTBefore antigen stimulation, most granules with CD63-GFP hardly moved in RBL cells. However, after antigen stimulation, the granules moved dramatically. They reached the plasma membranes in a few minutes and fused with them instantaneously. The velocity of the granule movement toward the plasma membranes on antigen stimulation was calculated to be 0.05 micron x s(-1).
CONCLUSIONAnalysis of the movement of each granule provided a new insight into the elementary process of degranulation. The method is rapid, sensitive and reliable, which could be used as a new detection method for anaphylactoid reaction in vitro.
