Boluda L, Berrens L
Research Group C.B.F. LETI, Tres Cantos, Madrid, Spain.
Clin Exp Immunol. 1995 Apr;100(1):145-50. doi: 10.1111/j.1365-2249.1995.tb03616.x.
IgG anti-IgE autoantibodies in human allergic sera have been investigated using an enzyme immunoassay on microplates coated with anti-human IgE MoAb. After incubation with serum, the plates were developed with peroxidase-labelled anti-human IgE MoAb for the determination of total IgE levels, or with anti-IgG MoAb for evaluating IgG anti-IgE autoantibodies. Using this methodology, no correlation was found between total IgE and IgG anti-IgE levels in groups of sera of allergic individuals. Although the results obtained with enzyme-labelled anti-IgG are often interpreted as indicative of IgE-IgG complexes captured from the serum, molecular sieving on gel columns as well as direct ultrafiltration experiments through 300-kD membranes demonstrate that such complexes do not occur preformed in the circulation, but arise de novo on the anti-IgE-coated solid phase during in vitro incubation with human serum. It is suggested that IgG anti-IgE autoantibodies react with IgE only after the latter has undergone a conformational change, either by colloidal manipulation or after reaction with allergen.
利用酶免疫测定法,在包被有抗人IgE单克隆抗体的微孔板上对人过敏性血清中的IgG抗IgE自身抗体进行了研究。血清孵育后,微孔板用辣根过氧化物酶标记的抗人IgE单克隆抗体显色以测定总IgE水平,或用抗IgG单克隆抗体显色以评估IgG抗IgE自身抗体。使用这种方法,在过敏性个体血清组中未发现总IgE与IgG抗IgE水平之间存在相关性。尽管用酶标记的抗IgG获得的结果通常被解释为表明从血清中捕获的IgE-IgG复合物,但凝胶柱上的分子筛以及通过300-kD膜的直接超滤实验表明,此类复合物并非在循环中预先形成,而是在与人类血清进行体外孵育期间在抗IgE包被的固相上重新形成。有人提出,IgG抗IgE自身抗体仅在IgE经历构象变化后才与之反应,这种构象变化要么是通过胶体操作,要么是在与变应原反应之后发生。