Harchali A A, Montagne P, Ruf J, Cuillière M L, Bene M C, Faure G, Duheille J
Immunology Laboratory, Faculty of Medicine, Vandoeuvre, France.
Clin Chem. 1994 Mar;40(3):442-7.
Crude thyroid peroxidase extracted from human thyroid microsomes was covalently bound onto polyacrylic and polyfunctional copolymerized microparticles. We observed agglutination of the thyroid peroxidase-microparticle conjugate with 13 monoclonal antibodies (mAbs) specific for epitopes on four different antigenic domains of human thyroid peroxidase (TPO; EC 1.11.1.7), after addition of anti-mouse immunoglobulins. We quantified agglutination by measuring with a specially designed nephelometer the light scattered by the conjugates. This allowed us to develop a microparticle-enhanced nephelometric immunoassay for human anti-TPO autoantibodies (aAbs) with defined epitopic specificity, based on the ability of aAbs to inhibit mAb-induced agglutination. Applied to patients with autoimmune thyroid diseases, this assay confirmed the polyclonality of anti-TPO aAbs and their preferential reactivity toward epitopes located on the A and B antigenic domains of the TPO molecule. The same specificities seem to be present in patients with Hashimoto thyroiditis or Graves disease.
从人甲状腺微粒体中提取的粗甲状腺过氧化物酶被共价结合到聚丙烯酸和多功能共聚微粒上。加入抗小鼠免疫球蛋白后，我们观察到甲状腺过氧化物酶 - 微粒偶联物与针对人甲状腺过氧化物酶（TPO；EC 1.11.1.7）四个不同抗原结构域上的表位的13种单克隆抗体（mAb）发生凝集。我们通过用专门设计的散射比浊仪测量偶联物散射的光来量化凝集。这使我们能够基于抗TPO自身抗体（aAb）抑制mAb诱导的凝集的能力，开发一种针对具有确定表位特异性的人抗TPO自身抗体的微粒增强散射比浊免疫测定法。应用于自身免疫性甲状腺疾病患者，该测定法证实了抗TPO aAb的多克隆性及其对TPO分子A和B抗原结构域上的表位的优先反应性。桥本甲状腺炎或格雷夫斯病患者似乎也存在相同的特异性。
