Calderon R A, Shennan G I
Department of Medical Microbiology, London School of Hygiene and Tropical Medicine, UK.
J Gen Microbiol. 1987 Sep;133(9):2689-98. doi: 10.1099/00221287-133-9-2689.
A monoclonal antibody (Tq-1) that interacts with the phosphorylcholine (PC)-bearing antigens of Trichophyton quinckeanum was produced by fusion of myeloma cells (JKAg-8) with spleen cells of BALB/c mice immunized with an alum-precipitated fraction of T. quinckeanum cytoplasmic antigen. It was characterized as an IgM class antibody by immunodiffusion using anti-Ig heavy chain specific reagents, ELISA using immunoglobulin-specific peroxidase-conjugated antibodies, and by gel filtration chromatography; it showed high affinity for Staphylococcus aureus protein-A. Interaction of Tq-1 with PC-like antigens of T. quinckeanum was demonstrated by inhibition studies using ELISA, immunoblotting, immunoprecipitation and immuno electron microscopy techniques. The binding activity of Tq-1 antibody with a range of dermatophyte proteins was completely inhibited by prior incubation with PC hapten. Moreover, dermatophyte antigens reacting with the monoclonal antibody reacted strongly with sera from chronically infected mice. Dermatophyte antigens derived from both young (24 h) and old (20 d) cultures reacted with Tq-1 and this binding was inhibited by PC, suggesting that Tq-1 target antigen PC appears at an early stage during fungal growth and remains throughout its life.
通过将骨髓瘤细胞（JKAg - 8）与用明矾沉淀的须发癣菌胞质抗原免疫的BALB/c小鼠脾细胞融合，制备了一种与须发癣菌的含磷酰胆碱（PC）抗原相互作用的单克隆抗体（Tq - 1）。通过使用抗Ig重链特异性试剂的免疫扩散、使用免疫球蛋白特异性过氧化物酶偶联抗体的ELISA以及凝胶过滤色谱法，将其鉴定为IgM类抗体；它对金黄色葡萄球菌蛋白A表现出高亲和力。使用ELISA、免疫印迹、免疫沉淀和免疫电子显微镜技术进行的抑制研究证明了Tq - 1与须发癣菌的PC样抗原的相互作用。Tq - 1抗体与一系列皮肤癣菌蛋白的结合活性在与PC半抗原预先孵育后被完全抑制。此外，与单克隆抗体反应的皮肤癣菌抗原与慢性感染小鼠的血清强烈反应。来自年轻（24小时）和年老（20天）培养物的皮肤癣菌抗原均与Tq - 1反应，并且这种结合被PC抑制，这表明Tq - 1的靶抗原PC在真菌生长的早期阶段出现并在其整个生命周期中持续存在。
