Mühlebach Michael D, Hutzler Stefan
Product Testing of IVMP, Division Veterinary Medicine, Paul-Ehrlich-Institut, Paul-Ehrlich-Str. 51-59, 63225, Langen, Germany.
Methods Mol Biol. 2017;1581:151-168. doi: 10.1007/978-1-4939-6869-5_9.
This chapter describes the development of recombinant measles virus (MV)-based vaccines starting from plasmid DNA. Live-attenuated measles vaccines are very efficient and safe. Since the availability of a reverse genetic system to manipulate MV genomes and to generate respective recombinant viruses, a considerable number of recombinant viruses has been generated that present antigens of foreign pathogens during MV replication. Thereby, robust humoral and cellular immune responses can be induced, which have shown protective capacity in a substantial number of experiments.For this purpose, the foreign antigen-encoding genes are cloned into additional transcription units of plasmid based full-length MV vaccine strain genomes, which in turn are used to rescue recombinant MV by providing both full-length viral RNA genomes respective anti-genomes together with all protein components of the viral ribonucleoprotein complex after transient transfection of the so-called rescue cells. Infectious centers form among these transfected cells, which allow clonal isolation of single recombinant viruses that are subsequently amplified, characterized in vitro, and then evaluated for their immunogenicity in appropriate preclinical animal models.
本章描述了从质粒DNA开始基于重组麻疹病毒（MV）的疫苗的开发。减毒活麻疹疫苗非常有效且安全。自从有了用于操纵MV基因组并产生相应重组病毒的反向遗传系统以来，已经产生了大量的重组病毒，这些病毒在MV复制过程中呈现外来病原体的抗原。由此，可以诱导强大的体液免疫和细胞免疫反应，这在大量实验中已显示出保护能力。为此，将编码外来抗原的基因克隆到基于质粒的全长MV疫苗株基因组的额外转录单元中，在对所谓的拯救细胞进行瞬时转染后，通过提供全长病毒RNA基因组和相应的反基因组以及病毒核糖核蛋白复合体的所有蛋白质成分，进而用于拯救重组MV。在这些转染细胞中形成感染中心，这允许对单个重组病毒进行克隆分离，随后进行扩增、体外鉴定，然后在适当的临床前动物模型中评估其免疫原性。
