Choi E Y, Kim K J, Jeon K W
Department of Zoology, University of Tennessee, Knoxville 37996.
J Protozool. 1992 Nov-Dec;39(6):671-7. doi: 10.1111/j.1550-7408.1992.tb04447.x.
Monoclonal antibodies were prepared against lysosomal membrane proteins of amoebae and used to follow lysosome-phagosome fusion after induced phagocytosis. The specificity of antibodies was checked by indirect immunofluorescence microscopy, immunoelectron microscopy, and localization of the antigen in subcellular fractions. The antibody-recognized proteins started to appear on the membranes of phagolysosomes about 5 min after phagocytosis as detected by indirect immunofluorescence, and the intensity of fluorescence increased for up to 1 h. Results of injection experiments in which purified antibodies had been injected into living cells and probed by indirect fluorescence indicated that the antigens were located on the cytoplasmic side of the lysosomal membranes. Lysosomes fuse with phagosomes on the one hand but not with non-fusible vesicles such as symbiosomes on the other. The results support the view that a membrane component(s) of non-fusible vesicles somehow prevents lysosomes from fusing with them.
制备了针对变形虫溶酶体膜蛋白的单克隆抗体，并用于追踪诱导吞噬作用后溶酶体-吞噬体的融合。通过间接免疫荧光显微镜、免疫电子显微镜以及抗原在亚细胞组分中的定位来检查抗体的特异性。通过间接免疫荧光检测，抗体识别的蛋白在吞噬作用后约5分钟开始出现在吞噬溶酶体的膜上，并且荧光强度持续增加长达1小时。将纯化抗体注射到活细胞中并通过间接荧光进行检测的注射实验结果表明，抗原位于溶酶体膜的细胞质一侧。一方面，溶酶体与吞噬体融合，但另一方面不与诸如共生体等不可融合的囊泡融合。这些结果支持这样一种观点，即不可融合囊泡的一种膜成分以某种方式阻止溶酶体与它们融合。
