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アナフィラキシ-関連物質の免疫測定による証明

通过免疫测定证明过敏反应相关物质

基本信息

批准号：
01570344
负责人：
KIMURA Hiroko
金额：
$ 1.22万
依托单位：
Juntendo University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

1) Quantitation of histamine.A new immunologic method to measure histamine concentration in blood has been developed. The principle of this method is based on the planimetric measurement of the agglutinated and precipitated area on a microtiter plate produced by heavy latex particles. The sensitivity of this assay was 0.1 ng/ml(= lnM) for histamine, and the coefficients of variation were 0.43-8.8% at 7 different concentrations (n=4). No cross-reactivity occurred with histidine, 1-methyl histamine or norepinephrine. A good correlation with the radioimmunoassay was obtained for rabbit blood, in which histamine was released either by immunogen stimulation or injection of compound 48/80 (r=0.977, n=26).2) Detection of allergic antibody to tick."A continuously rotating passive ag, -, , Iutination method" was carried out by using a newly developed immunologic carrier particle to detect allergic antibody to tick. This test showed a higher detection rate than the radioallergosorbent test.3) Morphologic comparison of anti-tick antibodies of different immunoglobulin classes.We also investigated the conformation of allergen adsorbed latex agglutinates with a scannin-, electron microscope. The antigen-antibody complexes incorporating IgE antibodies tend to form filaments whereas those containing IgG or ICm antibodies formed short connecting structures. This difference is probably contributed to the high elasticity of IgE antibody, and is evidence of the high biologic activity of the IgE antibody.4) Quantitation of human total IgE by the photothermal deflection spectrometry.A serum sample containing & IgE was stained on a nitrocellulose film, and anti-IgE rabbit IOG and anti-rabbit IgG coated gold particles were consecutively adsorbed to the film. The quantities of gold particles adsorbed were measured by the photothermal deflection method. Human IgE concentrations were determined from 0.076IU/ml to 76OIU/ml.

1)组胺的定量：建立了一种新的血中组胺浓度的免疫测定方法。该方法的原理是基于在微量滴定板上由重乳胶颗粒产生的凝集和沉淀面积的平面测量。本法对组胺的灵敏度为0.1ng/ml（= 1 nM），变异系数为0.43-8.8%。与组氨酸、1-甲基组胺或去甲肾上腺素无交叉反应。与免疫原刺激或注射化合物48/80释放组胺的兔血放射免疫测定结果均具有良好的相关性（r=0.977，n=26）。本文介绍了一种用新研制的免疫载体颗粒检测蜱过敏性抗体的“连续旋转被动抗原-- 3）不同免疫球蛋白类抗蜱抗体的形态学比较，并用扫描电镜观察吸附变应原乳胶凝集物的构象。掺入IgE抗体的抗原-抗体复合物倾向于形成细丝，而含有IgG或ICm抗体的抗原-抗体复合物形成短连接结构。这种差异可能与IgE抗体的高弹性有关，是IgE抗体高生物活性的证据。4）光热偏转光谱法定量人总IgE将含有IgE的血清样品在硝酸纤维素膜上染色，抗IgE兔IOG和抗兔IgG包被的金颗粒连续吸附在膜上。用光热偏转法测定了金粒子的吸附量。测定的人IgE浓度为0.076 IU/ml至760 IU/ml。