Thompson W S, Gross R H
Department of Biological Sciences, Dartmouth College, Hanover, NH 03755.
Gene Anal Tech. 1988 Jul-Aug;5(4):73-9. doi: 10.1016/0735-0651(88)90007-6.
We have developed a system for efficiently packaging antibodies and other macromolecules into liposomes and then delivering the encapsulated molecules into living cells through liposome-cell fusion. Fusion is very efficient, and all cells can be demonstrated to contain liposome-delivered antibodies by staining with a fluorescent second antibody. Using lupus antibodies directed against small nuclear ribonucleoprotein components of the cell, we were able to demonstrate strong nuclear localization, while control antibodies showed a general diffuse distribution throughout the cell. Lupus antibodies directed against ribosomes, on the other hand, strongly localized in the nucleolus and the cytoplasm with very little nucleoplasmic localization. Antitubulin antibodies predominantly localized in the cytoplasm. These results show that antibodies can survive liposome packaging and can retain their ability to recognize and bind to their specific antigens in the living cell. It also indicates that the nuclear envelope does not present a barrier to the liposome-introduced antibodies in Drosophila tissue culture cells. To determine if the antibodies were capable of interfering with cellular processes in vivo, we measured the effects of liposome-introduced antiribosome antibodies on translation and antitubulin antibodies on mitosis. In both cases, there was a significant inhibition suggesting that the antibodies can be used to interfere with specific functions at specific times in vivo.
我们开发了一种系统,可将抗体及其他大分子高效地包装到脂质体中,然后通过脂质体与细胞的融合将包封的分子递送至活细胞内。融合效率很高,通过用荧光二抗染色可证明所有细胞均含有脂质体递送的抗体。使用针对细胞小核核糖核蛋白成分的狼疮抗体,我们能够证明其强烈的核定位,而对照抗体则在整个细胞中呈现普遍的弥散分布。另一方面,针对核糖体的狼疮抗体强烈定位于核仁和细胞质,核质定位极少。抗微管蛋白抗体主要定位于细胞质。这些结果表明,抗体能够在脂质体包装后存活,并能在活细胞中保留识别和结合其特定抗原的能力。这也表明,在果蝇组织培养细胞中,核膜不会对脂质体引入的抗体构成屏障。为了确定这些抗体是否能够在体内干扰细胞过程,我们测量了脂质体引入的抗核糖体抗体对翻译的影响以及抗微管蛋白抗体对有丝分裂的影响。在这两种情况下,均有显著抑制作用,表明这些抗体可用于在体内特定时间干扰特定功能。