A fluorometric microassay for histamine release from human gingival mast cells.
Mast cells are important effector cells of the immune system. We describe a rapid and inexpensive microassay to determine histamine release from human gingival mast cells. The assay is based on the coupling of histamine with o-phthalaldehyde (OPT) at a highly alkaline pH to form a fluorescent product. Using this assay with a sample volume of 10 microl/well in a 384 black well microplate, the histamine detection limit was 0.031 microg/ml. The human mast cell line (HMC-1) and fresh mast cells isolated from human gingival tissue (n = 10) were stimulated with substance P, anti-IgE or calcium ionophore A23187. Calcium ionophore significantly increased histamine release from HMC-1 cells and gingival mast cells (p < 0.05). This microassay will facilitate the study of mast cell histamine release in diseased oral mucosa.
Zhao
Z Z
ZZ
School of Dentistry, University of Queensland, Brisbane, Australia.
Sugerman
P B
PB
Walsh
L J
LJ
Savage
N W
NW
D002478:Cells, Cultured; D005470:Fluorometry / Q000379:methods; D005881:Gingiva / Q000166:cytology*; D006632:Histamine / Q000032:analysis*; D006636:Histamine Release; D006801:Humans; D007202:Indicators and Reagents; D008407:Mast Cells / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric; D009764:o-Phthalaldehyde
D007202:Indicators and Reagents; D009764:o-Phthalaldehyde; D006632:Histamine