Jean-François M J, Poskitt D C, Turnbull S J, Macdonald L M, Yasmeen D
Commonwealth Serum Laboratories, Research and Development Division, Parkville, Victoria, Australia.
J Gen Microbiol. 1991 Sep;137(9):2125-33. doi: 10.1099/00221287-137-9-2125.
We have developed an in vivo passive transfer assay using mice to identify monoclonal antibodies (mAbs) which offer protection against Streptococcus equi infection. The assay was developed using serum antibodies collected from horses convalescing from strangles. In this study, we show that a preparation of M-like protein, acid-extracted from S. equi, affords 80% protection to mice immunized with it. A number of mouse mAbs directed against a preparation of M-like protein were then assessed for their ability to passively protect mice against challenge with a lethal dose of the bacteria. Two mAbs, 1D10 and 2A6, were shown to be highly protective. It was also demonstrated, by means of a competitive enzyme immunoassay, that these mAbs recognized different epitopes in the preparation. Examination of a dose-response curve for mAbs 1D10 and 2A6 revealed that optimal levels of protection were achieved using 1 mg of either 1D10 or 2A6, or 0.5 mg 1D10 and 0.5 mg 2A6 given together. Immunological reactivity of these mAbs with a preparation of M-like protein showed that the antigens they recognized were comparable in size to some of the antigens recognized by convalescent horse serum antibodies. The role of immunoglobulin isotype in conferring protection is discussed.
我们开发了一种利用小鼠的体内被动转移试验,以鉴定可提供抗马链球菌感染保护作用的单克隆抗体(mAb)。该试验是利用从患腺疫正在康复的马身上采集的血清抗体开发的。在本研究中,我们表明,从马链球菌中酸提取的M样蛋白制剂,能为用其免疫的小鼠提供80%的保护。然后评估了多种针对M样蛋白制剂的小鼠单克隆抗体被动保护小鼠免受致死剂量细菌攻击的能力。两种单克隆抗体1D10和2A6显示出高度的保护作用。通过竞争性酶免疫测定还证明,这些单克隆抗体识别制剂中的不同表位。对单克隆抗体1D10和2A6的剂量反应曲线进行检查发现,使用1mg的1D10或2A6,或一起给予0.5mg的1D10和0.5mg的2A6可达到最佳保护水平。这些单克隆抗体与M样蛋白制剂的免疫反应性表明,它们识别的抗原在大小上与患腺疫康复马血清抗体识别的一些抗原相当。讨论了免疫球蛋白同种型在提供保护中的作用。