Antibodies introduced into living cells with liposomes localize specifically and inhibit specific intracellular processes

W. Scott Thompson, Robert H. Gross
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引用次数: 9

Abstract

We have developed a system for efficiently packaging antibodies and other macromolecules into lipsomes 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 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.

通过脂质体引入活细胞的抗体可以特异地定位并抑制特定的细胞内过程
我们已经开发了一种系统,可以有效地将抗体和其他大分子包装到脂质体中,然后通过脂质体-细胞融合将被封装的分子输送到活细胞中。融合是非常有效的,所有细胞都可以通过荧光第二抗体染色来证明含有脂质体传递的抗体。使用针对细胞小核核糖核蛋白成分的狼疮抗体,我们能够证明强大的核定位,而对照抗体显示整个细胞的普遍弥漫性分布。另一方面,针对核糖体的狼疮抗体强烈定位于核核和细胞质中,核质定位很少。抗微管蛋白抗体主要存在于细胞质中。这些结果表明,抗体可以在脂质体包装中存活,并在活细胞中保留其识别和结合特异性抗原的能力。这也表明在果蝇组织培养细胞中,核膜对脂质体引入的抗体不存在屏障。为了确定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和抗微管蛋白抗体对有丝分裂的影响。在这两种情况下,都有明显的抑制作用,表明抗体可以在体内特定时间干扰特定功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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