DETECTION OF ANTIGEN-INDUCED HISTAMINE RELEASE IN WHOLE PERIPHERAL BLOOD FROM ASHMATIC CHILDREN

A COMPARATIVE STUDY BETWEEN HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AND RADIOIMMUNOASSAY

Abstract

Mite antigen-induced histamine release was examined using whole peripheral blood from asthmatic patients. Released histamine concentrations were determined by both high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) and the values were compared stastistically.
Samples were taken from 9 children (5 males and 4 females; Age 6-13 years) with IgE RAST Score 4 positive to housedust mite, Dermatophagoides farinae.
Three different histamine levels, histamine release challenged with mite antigen at concentrations ranging from 10^-4 to 10⁰μg/ml, spontaneous histamine release, and total histamine levels taken by repeated freeze/thawing, were measured by both HPLC and RIA.
There was a strong correlation (r=0.948, n=63, p<0.001) between the logarithm of values determined by RIA and those determined by HPLC. In addition, the RIA method needs only about 1ml of whole peripheral blood.
Since the reliability of histamine level detrmination by HPLC is established, these results indicate that the measurement of histamine release in vitro by RIA is sufficiently reliable, simple, and clinically beneficial for young ashmatic children.