A new method for detecting histamine release.

Abstract

Glass microfibres have been found to bind histamine with high affinity and selectivity. A new test for measuring basophil histamine release has been developed using the glass microfibres as a solid phase. Glass microfibres are crushed and fixed to the bottom of microtitre plates with a water-soluble glue. Histamine release is performed in the glass microfibre -prepared microtitre plates by challenging 100 microliter washed blood with 20 microliter antigen per well for 90 min at 37 degrees C. Released histamine is bound with high affinity to the glass microfibres , since 90% of histamine in the solution is adsorbed to the fibres. After incubation the microtitre plate is washed with H2O to remove cells and interfering substances. Fibre-bound histamine is detected by the fluorometric o-phthaldialdehyde method. The sensitivity of the assay is 0.63 ng histamine, 2 HCl and the histamine standard curve is linear up to at least 5 ng histamine, 2 HCl. Optimal conditions for the new assay are described. After challenge with anti-Ige a comparison with the conventional histamine release from Ficoll-Hypaque-isolated leukocytes showed almost identical results.