Moore R J, Lenghaus C, Sheedy S A, Doran T J
Australian Animal Health Laboratory, CSIRO Livestock Industries, Private Bag 24, Vic. 3220, Geelong, Australia.
Vaccine. 2001 Oct 12;20(1-2):115-20. doi: 10.1016/s0264-410x(01)00314-0.
Expression library immunization (ELI) has previously been used in a number of disease models in mice. Here, we describe the first example of the application of ELI to a large animal model with the immunization of pigs against enzootic pneumonia, a disease caused by Mycoplasma hyopneumoniae. The development of new plasmid vectors and library screening methods facilitated the application of ELI to this disease by allowing random libraries to be screened for clones expressing recombinant proteins. In this way the vast majority of clones in random libraries that are unproductive can be eliminated, meaning that libraries are more likely to give protection and are subsequently easier to further screen and analyze. By using this approach we have used one library screen and two animal trials to progress from an original library of 20,000 clones to a group of just 96 clones.
表达文库免疫(ELI)此前已在多种小鼠疾病模型中使用。在此,我们描述了ELI应用于大型动物模型的首个实例,即对猪进行免疫以预防由猪肺炎支原体引起的地方流行性肺炎。新质粒载体和文库筛选方法的开发,通过对随机文库筛选表达重组蛋白的克隆,促进了ELI在该疾病中的应用。通过这种方式,可以消除随机文库中绝大多数无生产性的克隆,这意味着文库更有可能提供保护,随后更易于进一步筛选和分析。通过使用这种方法,我们通过一次文库筛选和两次动物试验,从最初的20000个克隆文库进展到仅96个克隆的一组。