Lentiviral transduction of Tar Decoy and CCR5 ribozyme into CD34+ progenitor cells and derivation of HIV-1 resistant T cells and macrophages.
Lentiviral transduction of Tar Decoy and CCR5 ribozyme into CD34+ progenitor cells and derivation of HIV-1 resistant T cells and macrophages.
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DOI：
10.1186/1742-6405-1-2
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发表时间：
2004-12-17
影响因子：
2.2
通讯作者：
Akkina, Ramesh
中科院分区：
文献类型：
--
作者：
Banerjea, Akhil;Li, Ming-Jie;Akkina, Ramesh
背景：基于 RNA 的 HIV-1 抗病毒方法是最有前途的长期基因治疗方法之一。这些包括核酶、适体（诱饵）和小干扰 RNA (siRNA)。慢病毒载体非常适合将此类抑制性 RNA 转导至造血干细胞，因为它们能够转导非分裂细胞，并且对基因沉默具有相对折射性。本研究的目的是通过基于 HIV 的慢病毒载体将 HIV-1 Tar 适体单独或与抗 CCR5 核酶组合引入 CD34+ 造血祖细胞，以获得病毒抗性子代 T 细胞和巨噬细胞。结果：使用含有 Tar 诱饵或 Tar 诱饵与 CCR5 核酶组合的慢病毒载体构建体，实现了向 CD34+ 细胞的高效且持续的基因转移。用这些构建体转导的细胞在 SCID-hu 小鼠的 thy/liv 移植物中正常体内分化为 T 淋巴细胞，并在存在适当生长因子的情况下在体外正常分化为巨噬细胞。当在体外受到攻击时，分化的T淋巴细胞和巨噬细胞表现出对HIV-1感染的显着抵抗力。结论：通过CD34+祖细胞的慢病毒基因转导可以获得单独表达HIV-1 Tar适体或与抗CCR5核酶组合表达HIV-1 Tar适体的病毒抗性转基因T细胞和巨噬细胞。这些结果首次表明，这些抗HIV-1转基因的组合表达不会干扰正常的胸腺生成，因此为它们在基于干细胞的HIV/AIDS基因治疗中的应用奠定了基础。
BACKGROUND: RNA based antiviral approaches against HIV-1 are among the most promising for long-term gene therapy. These include ribozymes, aptamers (decoys), and small interfering RNAs (siRNAs). Lentiviral vectors are ideal for transduction of such inhibitory RNAs into hematopoietic stem cells due to their ability to transduce non-dividing cells and their relative refractiveness to gene silencing. The objective of this study is to introduce an HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme into CD34+ hematopoietic progenitor cells via an HIV-based lentiviral vector to derive viral resistant progeny T cells and macrophages. RESULTS: High efficiency and sustained gene transfer into CD34+ cells were achieved with lentiviral vector constructs harboring either Tar decoy or Tar decoy in combination with CCR5 ribozyme. Cells transduced with these constructs differentiated normally into T-lymphocytes in vivo in thy/liv grafts of SCID-hu mice, and into macrophages in vitro in the presence of appropriate growth factors. When challenged in vitro, the differentiated T lymphocytes and macrophages showed marked resistance against HIV-1 infection. CONCLUSIONS: Viral resistant transgenic T cells and macrophages that express HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme could be obtained by lentiviral gene transduction of CD34+ progenitor cells. These results showed for the first time that expression of these anti-HIV-1 transgenes in combination do not interfere with normal thymopoiesis and thus have set the stage for their application in stem cell based gene therapy for HIV/AIDS.