TAT-MEDIATED DELIVERY OF HETEROLOGOUS PROTEINS INTO CELLS
TAT-MEDIATED DELIVERY OF HETEROLOGOUS PROTEINS INTO CELLS
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DOI：
10.1073/pnas.91.2.664
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发表时间：
1994-01-18
影响因子：
11.1
通讯作者：
BARSOUM, J
中科院分区：
文献类型：
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作者：
FAWELL, S;SEERY, J;BARSOUM, J
人类免疫缺陷病毒1型(HIV-1)的Tat蛋白在组织培养中加入外源能有效进入细胞。为了评估TAT是否可以携带其他分子进入细胞，我们将TAT多肽(残基1-72或37-72)化学交联到假单胞菌外毒素A(PE)的β-半乳糖苷酶、辣根过氧化物酶、核糖核酸酶A和结构域III，并通过比色法或细胞毒性监测摄取。TAT嵌合体对所有测试的细胞类型都有效，染色显示在每个实验中都吸收了所有的细胞。在小鼠中，用TAT-β-半乳糖苷酶嵌合体治疗后，可以将药物输送到几个组织中，其中心脏、肝脏和脾中的含量较高，肺和骨骼肌中的含量较低到中等，而肾脏和大脑中的含量很少或没有活性。这些组织内的主要靶点是血管周围的细胞，即内皮细胞、库普弗细胞和/或脾巨噬细胞。TAT介导的摄取可能允许以前被认为不能渗透到活细胞的大分子的治疗性输送。
The Tat protein of human inmunodeficiency virus 1 (HIV-1) can enter cells efficiently when added exogenously in tissue culture. To assess if Tat can carry other molecules into cells, we chemically cross-linked Tat peptides (residues 1-72 or 37-72) to beta-galactosidase, horseradish peroxidase, RNase A, and domain III of Pseudomonas exotoxin A (PE) and monitored uptake colorimetrically or by cytotoxicity. The Tat chimeras were effective on all cell types tested, with staining showing uptake into all tells in each experiment. In mice, treatment with Tat-beta-galactosidase chimeras resulted in delivery to several tissues, with high levels in heart, liver, and spleen, low to-moderate levels in lung and skeletal muscle, and little or no activity in kidney and brain. The primary target within these tissues was the cells surrounding the blood vessels, suggesting endothelial cells, Kupffer cells, and/or splenic macrophages. Tat mediated uptake may allow the therapeutic delivery of macromolecules previously thought to be impermeable to living cells.