The efficient and rapid import of a peptide into primary B and T lymphocytes and a lymphoblastoid cell line
The efficient and rapid import of a peptide into primary B and T lymphocytes and a lymphoblastoid cell line
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DOI：
10.1016/s0022-1759(97)00208-1
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发表时间：
1998-03-01
影响因子：
2.2
通讯作者：
Sinclair, AJ
中科院分区：
文献类型：
--
作者：
Fenton, M;Bone, N;Sinclair, AJ
淋巴细胞是出了名的难以转化。受欢迎的电穿孔技术有三个主要缺点：它具有高度破坏性，导致大规模细胞死亡；它效率低下；静止的初级淋巴细胞除非被部分激活，否则不能进行电穿孔。我们已经研究了触角线虫同源结构域蛋白(PAntp)的第三螺旋的细胞导入作为一种将多肽引入原代淋巴细胞和淋巴细胞系的替代方法。PAntp多肽被迅速吸收到细胞质和细胞核中，并在那里保留至少48小时。该系统没有表现出电穿孔的缺点；在所测试的浓度下没有检测到pAntp多肽的毒性，该过程是有效的，高达95%的淋巴细胞导入pAntp多肽。最后，在导入pAntp多肽时，静止的初级淋巴细胞与激活的初级淋巴细胞和淋巴样细胞株的效率相同。这表明，pAntp多肽递送系统作为一种将分子递送到淋巴细胞和淋巴样细胞系的方法，比电穿孔具有重大优势。(C)1998 Elsevier Science B.V.保留所有权利。
Lymphocytes are notoriously difficult to transfect. The favoured technique, electroporation, has three major disadvantages: it is highly disruptive, causing large scale cell death; it is inefficient; quiescent primary lymphocytes are refractory to electroporation unless they have been partially activated. We have investigated the cellular import of the third helix of the Antennapedia homeodomain protein (pAntp) as an alternative method for introducing peptides into primary lymphocytes and lymphoid cell lines. The pAntp peptide is taken up rapidly into the cytoplasm and nucleus of the cells where it is retained for at least 48 h. The system displays none of the disadvantages of electroporation; no toxicity of the pAntp peptide was detected at the concentrations tested and the process was efficient with up to 95% of lymphocytes importing the pAntp peptide. Finally, quiescent primary lymphocytes were as efficient as activated primary lymphocytes and a lymphoid cell line at importing the pAntp peptide. This demonstrates that the pAntp peptide delivery system has major advantages over electroporation as a method of delivering molecules to lymphocytes and a lymphoid cell line. (C) 1998 Elsevier Science B.V. All rights reserved.