Ehrnst A, Sundqvist K G
Cell. 1975 Aug;5(4):351-9. doi: 10.1016/0092-8674(75)90053-7.
Fixation with glutaraldehyde (GA) and paraformaldehyde (PFA) preserved measles virus hemagglutinin at the surface of chronically infected cells. Cells fixed with PFA but not with GA exhibited hemadsorption with green monkey cells. PFA fixation, in contrast to GA fixation, also preserved the immunogenicity of measles virus hemolysin. These fixatives and the removal of the measles virus hemagglutinin from the cell surface by trypsin enabled studies of the appearance of the hemagglutinin at the surface membrane. Results obtained by immunofluorescence technique and by hemadsorption indicated that measles virus hemagglutinin appeared polarly at the cell membrane and then spread around the surface. This was substantiated by measurements of the immunofluorescence intensity at the single cell level per membrane unit and per cell, and by measuring the binding of iodinated immunoglobulins per 10(6) cells. The appearance was inhibited by sodium azide and cytochalasin B. The spreading was not inhibited by sodium azide, but was influenced by cytochalasin B. The spreading did not proceed at 4 degrees C. On the basis of these findings, a hypothetical model for appearance and spreading of measles virus hemagglutinin was proposed.
用戊二醛(GA)和多聚甲醛(PFA)固定可保存慢性感染细胞表面的麻疹病毒血凝素。用PFA而非GA固定的细胞与绿猴细胞表现出血细胞吸附作用。与GA固定不同,PFA固定还保留了麻疹病毒溶血素的免疫原性。这些固定剂以及用胰蛋白酶从细胞表面去除麻疹病毒血凝素,使得能够研究血凝素在细胞膜表面的出现情况。通过免疫荧光技术和血细胞吸附获得的结果表明,麻疹病毒血凝素在细胞膜上呈极性出现,然后在表面扩散。这通过在单细胞水平上测量每个膜单位和每个细胞的免疫荧光强度,以及测量每10⁶个细胞中碘化免疫球蛋白的结合情况得到了证实。这种出现受到叠氮化钠和细胞松弛素B的抑制。扩散不受叠氮化钠抑制,但受细胞松弛素B影响。在4℃时扩散不发生。基于这些发现,提出了一个关于麻疹病毒血凝素出现和扩散的假设模型。