Rolle der Transmembran-Segmente von Masern-Glykoproteinen im Aufbau der viralen Hülle und der Gebrauch fremder Glykoproteine auf Masernviren für eine divalente Impfung

了解支持囊膜组装的相互作用对于开发基于麻疹病毒(MV)的多价疫苗和载体靶向具有重要意义。根据情况的不同，可能会或不会考虑将外源蛋白纳入MV包膜。与分泌或在细胞膜上表达相同的蛋白相比，呈现规则排列在病毒颗粒中的外源糖蛋白可能会诱导更强大的免疫反应。我们先前已经证明，MV融合蛋白(F)和血凝素(H)蛋白细胞质尾部中的膜-远端氨基酸调节这些蛋白在病毒颗粒中的效率(Capomen等人，J.Virol)。1998年)。我们也有迹象表明，这些糖蛋白的跨膜(TM)片段可能有助于有效的颗粒掺入(M.Rager等人，未发表)。在这项工作中，我们的目的是确定F和H蛋白中对包膜掺入重要的残基，并验证所识别的氨基酸是否对选定的外源糖蛋白掺入MV颗粒有影响。理想情况下，这些实验将产生一组TM和细胞质尾部突变体，这些突变体是“便携的”，可以用来提高将外源糖蛋白掺入MV颗粒的分级效率。在第三组实验中，我们计划测量表达不同配置的报告糖蛋白的重组MVS所诱导的免疫反应的大小：分泌的、展示在病毒颗粒上的或在细胞表面的。

Understanding the interactions supporting envelope assembly is of central importance for the development of multivalent measles virus (MV)-based vaccines and for vector targeting. Depending on the circumstances, the incorporation of foreign proteins in the MV envelope may or not be envisaged. Presentation of a foreign glycoprotein regularly arranged in a viral particle may induce a more robust immune response than secretion of the same protein or its expression on the cell membrane. We have previously shown that the membrane-distal amino acids in the MV fusion (F) and hemagglutinin (H) protein cytoplasmic tail modulate the efficiency of these proteins in viral particles (Cathomen et al., J. Virol. 1998). We also have indications that the transmembrane (TM) segment of these glycoproteins may contribute to efficient particle incorporation (M.Rager et al., unpublished). In this work we aim at identifying the residues in the F and H proteins important for envelope incorporation, and to vrify whether the identified amino acids have an effect on the incorporation of selected foreign glycoproteins into MV particles. Ideally, these experiments will result in the production of a set of TM and cytoplasmic tail mutants that are "portable" and can be used to confer graded efficieny of incorporation of foreign glycoproteins into MV particles. In a third set of experiments, we plan to measure the magnitude of the immune response induced by recombinant MVs expressing a reporter glycoprotein in different configurations: secreted, displayed on viral particles, or on the cell surface.