Granato D A, Piguet P F
Clin Exp Immunol. 1986 Mar;63(3):703-10.
A monoclonal IgE antibody directed against bovine milk beta-lactoglobulin (beta-LG) was produced by fusion of NSI myeloma cells with spleen cells of Balb/c mice immunized with alum-precipitated beta-LG. This antibody was found by radioimmunoassay to react with both native and aggregated beta-LG, but a positive passive cutaneous anaphylaxis reaction (PCA) was evident only with aggregated beta-LG. 1 ng of this purified antibody was capable of eliciting a PCA. The chemical and physical properties were characterized by amino acid and carbohydrate analysis and by ultracentrifugation. The epsilon-chain had an apparent molecular weight of 86,000 +/- 2,000 by polyacrylamide gel electrophoresis. An immediate hypersensitivity reaction within the gut was elicited by intravenous (i.v.) administration of the hybridoma ascitic fluid followed by feeding with aggregated beta-LG. Accumulation of liquid within the small intestine and diarrhoea were evident 30-90 min later. Intravenous injection of carbon particles revealed an increased permeability of the venulae from the submucosa and serosa. Histological examination showed oedema within the villae, without modification of the epithelium.
通过将NSI骨髓瘤细胞与用明矾沉淀的β-乳球蛋白免疫的Balb/c小鼠的脾细胞融合,制备了一种针对牛乳β-乳球蛋白(β-LG)的单克隆IgE抗体。通过放射免疫测定发现该抗体与天然和聚集的β-LG均发生反应,但仅与聚集的β-LG有明显的阳性被动皮肤过敏反应(PCA)。1 ng这种纯化抗体能够引发PCA。通过氨基酸和碳水化合物分析以及超速离心对其化学和物理性质进行了表征。通过聚丙烯酰胺凝胶电泳,ε链的表观分子量为86,000±2,000。静脉内(i.v.)注射杂交瘤腹水,随后喂食聚集的β-LG,可引发肠道内的速发型超敏反应。30 - 90分钟后,小肠内液体蓄积和腹泻明显。静脉注射碳颗粒显示黏膜下层和浆膜层的小静脉通透性增加。组织学检查显示绒毛内有水肿,上皮未发生改变。