Laboratory of Lymphocyte Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
J Immunol Methods. 2010 Oct 31;362(1-2):190-4. doi: 10.1016/j.jim.2010.08.009. Epub 2010 Aug 26.
The African trypanosome (Trypanosoma brucei) is transmitted by the bite of the tsetse vector to the mammalian bloodstream where it exists as a completely extracellular parasite. As a result of this exposure, the parasite elicits a robust immune response that is almost exclusively antibody mediated, and is extremely specific to the trypanosome coat displayed on the surface. This coat is comprised of ~11 million copies of a single gpi-linked molecule (the variable surface glycoprotein or VSG) and can therefore be used as a powerful platform for the immunogenic display of antigenic determinants. Here we describe a method to display repetitive, ordered arrays of linear epitopes on the surface of T. brucei and to then use the engineered organisms to generate specific anti-epitope antibody responses, upon injection into mice. This method offers an alternative approach to generating anti-peptide antibodies, and could be a useful option in cases where more traditional methods have failed.
非洲锥虫(Trypanosoma brucei)通过采采蝇的叮咬传播到哺乳动物的血液中,在血液中它作为一种完全细胞外寄生虫存在。由于这种暴露,寄生虫引发了强烈的免疫反应,几乎完全是抗体介导的,并且对表面展示的锥虫涂层具有极高的特异性。这种涂层由约 1100 万个单糖基化磷脂酰肌醇(GPI)连接的分子(可变表面糖蛋白或 VSG)组成,因此可以用作免疫原性展示抗原决定簇的强大平台。在这里,我们描述了一种在 T. brucei 表面展示重复、有序的线性表位阵列的方法,然后使用工程化的生物体在注射到小鼠中后产生针对特定表位的抗体反应。该方法提供了一种产生抗肽抗体的替代方法,在传统方法失败的情况下可能是一个有用的选择。