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

了解支持包膜组装的相互作用对于开发基于多价麻疹病毒（MV）的疫苗和载体靶向至关重要。根据具体情况，可以设想或不设想外源蛋白在MV包膜中的掺入。有规律地排列在病毒颗粒中的外源糖蛋白比分泌相同的蛋白或其在细胞膜上的表达能引起更强的免疫反应。我们之前已经证明，MV融合(F)和血凝素(H)蛋白细胞质尾部的膜端氨基酸调节这些蛋白在病毒颗粒中的效率（Cathomen et al.， J. Virol. 1998）。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结合（M.Rager等人，未发表）。在这项工作中，我们的目的是鉴定F和H蛋白中对包膜结合重要的残基，并验证鉴定的氨基酸是否对选定的外源糖蛋白进入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“便携式”的，可用于赋予外源糖蛋白结合到MV颗粒中的分级效率。在第三组实验中，我们计划测量表达不同结构的报告糖蛋白的重组mv诱导的免疫反应的大小：分泌的，显示在病毒颗粒上的，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transmembrane (TM) segment of these glycoproteins may contribute to efficient particle incorporation (M.Rager et al., unpublished).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.