Kolk A H, Ho M L, Klatser P R, Eggelte T A, Kuijper S, de Jonge S, van Leeuwen J
Clin Exp Immunol. 1984 Dec;58(3):511-21.
Thirty-two monoclonal antibodies (MoAb) to Mycobacterium tuberculosis H37Rv, M. bovis BCG and M. leprae were produced. The spleen cells of BALB/c mice immunized with sonicated or intact bacilli were fused with Sp2/0-Ag-14 myeloma cells. Many more antibody producing hybridomas were found when M. tuberculosis, rather than M. leprae, was used as the immunogen. The MoAb were characterized by an enzyme immunoassay and immunofluorescence on 16 mycobacterial species. The sodium dodecylsulphate polyacrylamide gel electrophoresis immunoperoxidase assay was used to determine the molecular weight of the antigens detected by the MoAb. Antigens of high, low and intermediate molecular weight were found. Some of the antigens were proteinaceous, others of a glycolipid nature. The immunofluorescence assay proved to be essential for the selection of MoAb since some MoAb reacted only in this assay and not in the enzyme immunoassay. The most specific clones were found in the fusions with spleen cells of mice immunized with intact rather than sonicated bacteria. One MoAb (F29-29) reacted only with M. tuberculosis H37Rv; one (F41-3) only with M. leprae and another (F29-45) reacted with M. tuberculosis and M. gastrii. Several MoAb only reacted with three mycobacterial species: M. tuberculosis, M. kansasii and M. gastrii. Others showed unique patterns of reactivity by enzyme immuno- and immunofluorescence assay. The potential use of the MoAb for the identification of mycobacteria and mycobacterial antigens is discussed.
制备了32种针对结核分枝杆菌H37Rv、牛分枝杆菌卡介苗和麻风分枝杆菌的单克隆抗体（MoAb）。用超声处理的或完整的杆菌免疫的BALB/c小鼠的脾细胞与Sp2/0-Ag-14骨髓瘤细胞进行融合。当使用结核分枝杆菌而非麻风分枝杆菌作为免疫原时，发现产生抗体的杂交瘤更多。通过酶免疫测定和对16种分枝杆菌的免疫荧光对这些单克隆抗体进行了表征。采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳免疫过氧化物酶测定法来确定单克隆抗体所检测抗原的分子量。发现了高分子量、低分子量和中等分子量的抗原。一些抗原是蛋白质性质的，其他的则是糖脂性质的。免疫荧光测定对于单克隆抗体的筛选被证明是必不可少的，因为一些单克隆抗体仅在该测定中反应，而在酶免疫测定中不反应。在与用完整细菌而非超声处理的细菌免疫的小鼠脾细胞融合中发现了最具特异性的克隆。一种单克隆抗体（F29-29）仅与结核分枝杆菌H37Rv反应；一种（F41-3）仅与麻风分枝杆菌反应，另一种（F29-45）与结核分枝杆菌和胃分枝杆菌反应。几种单克隆抗体仅与三种分枝杆菌反应：结核分枝杆菌、堪萨斯分枝杆菌和胃分枝杆菌。其他单克隆抗体通过酶免疫测定和免疫荧光测定显示出独特的反应模式。讨论了这些单克隆抗体在分枝杆菌和分枝杆菌抗原鉴定中的潜在用途。
