In vivo suppression of HIV by antigen specific T cells derived from engineered hematopoietic stem cells.
In vivo suppression of HIV by antigen specific T cells derived from engineered hematopoietic stem cells.
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DOI：
10.1371/journal.ppat.1002649
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发表时间：
2012
期刊：
影响因子：
6.7
通讯作者：
Zack JA
中科院分区：
文献类型：
--
作者：
Kitchen SG;Levin BR;Bristol G;Rezek V;Kim S;Aguilera-Sandoval C;Balamurugan A;Yang OO;Zack JA
HIV特异性细胞毒性T淋巴细胞（CTL）反应是控制体内病毒复制的关键组成部分，但最终无法根除病毒。我们在这些研究中的目的是开发增强和恢复 HIV 特异性 CTL 反应的方法，以实现长期病毒抑制或病毒清除。在我们的方法中，我们试图对人类造血干细胞 (HSC) 进行基因操作，使其分化成成熟的 CTL，从而杀死 HIV 感染的细胞。为此，我们从 CD8+ T 细胞中分子克隆了 HIV 特异性 T 细胞受体 (TCR)，该受体专门针对 HIV-1 Gag 蛋白的表位。然后该TCR被用于基因转导HSC。然后将这些 HSC 引入含有人胎儿肝脏、胎儿胸腺和造血祖细胞的人源化小鼠中，并使其分化为成熟的人 CD8+ CTL。我们在小鼠的多个组织中发现了人类 HIV 特异性 CTL。因此，用克隆的 TCR 对人类 HSC 进行基因修饰，可以使细胞在体内发生适当的分化，并且这些细胞可以迁移到多个解剖部位，模仿人类中所见的情况。为了确定转基因 HIV 特异性 TCR 的存在是否对抑制 HIV 复制有影响，我们感染了表达转基因 HIV 特异性 TCR 的 HIV-1 小鼠，并分别感染了表达非特异性对照 TCR 的小鼠。与对照组相比，我们观察到表达 HIV 特异性 TCR 的小鼠多个器官中 HIV 复制受到显着抑制，这表明转基因 HIV 特异性 CTL 的存在可以在体内形成功能性抗病毒反应。这些结果强烈表明，基于干细胞的基因治疗可能是治疗慢性病毒感染的可行方法，并为此类策略的发展奠定了基础。迫切需要开发新的治疗策略来根除艾滋病毒感染。 HIV 会积极破坏针对它的有效自然免疫反应，特别是细胞毒性 T 淋巴细胞 (CTL) 反应。开发一种疗法，使 HIV 能够长期免疫自我遏制，并由宿主恢复这些 CTL 反应，这将是理想的选择。通过对人类造血干细胞进行基因操作，我们引入了一种分子——一种针对 HIV 的 T 细胞受体 (TCR)——在人源化小鼠模型中，该分子可以在人体组织内分化后生成功能性 HIV 特异性 CTL。为了评估这些新开发的 HIV 特异性 CTL 是否可以主动抑制 HIV 复制，我们用 HIV 感染了这些小鼠。我们发现，这些小鼠中转基因的 HIV 特异性 CTL 的发育导致体内出现功能性抗病毒 CTL 反应，从而显着降低 HIV 感染后的病毒复制。这些结果对于使用该技术来设计人体免疫反应以对抗病毒感染具有重要意义，并表明通过造血干细胞进行基因工程可能允许调整免疫反应以瞄准和根除艾滋病毒。
The HIV-specific cytotoxic T lymphocyte (CTL) response is a critical component in controlling viral replication in vivo, but ultimately fails in its ability to eradicate the virus. Our intent in these studies is to develop ways to enhance and restore the HIV-specific CTL response to allow long-term viral suppression or viral clearance. In our approach, we sought to genetically manipulate human hematopoietic stem cells (HSCs) such that they differentiate into mature CTL that will kill HIV infected cells. To perform this, we molecularly cloned an HIV-specific T cell receptor (TCR) from CD8+ T cells that specifically targets an epitope of the HIV-1 Gag protein. This TCR was then used to genetically transduce HSCs. These HSCs were then introduced into a humanized mouse containing human fetal liver, fetal thymus, and hematopoietic progenitor cells, and were allowed to differentiate into mature human CD8+ CTL. We found human, HIV-specific CTL in multiple tissues in the mouse. Thus, genetic modification of human HSCs with a cloned TCR allows proper differentiation of the cells to occur in vivo, and these cells migrate to multiple anatomic sites, mimicking what is seen in humans. To determine if the presence of the transgenic, HIV-specific TCR has an effect on suppressing HIV replication, we infected with HIV-1 mice expressing the transgenic HIV-specific TCR and, separately, mice expressing a non-specific control TCR. We observed significant suppression of HIV replication in multiple organs in the mice expressing the HIV-specific TCR as compared to control, indicating that the presence of genetically modified HIV-specific CTL can form a functional antiviral response in vivo. These results strongly suggest that stem cell based gene therapy may be a feasible approach in the treatment of chronic viral infections and provide a foundation towards the development of this type of strategy. There is a desperate need for the development of new therapeutic strategies to eradicate HIV infection. HIV actively subverts the potent natural immune responses against it, particularly cellular cytotoxic T lymphocyte (CTL) responses. The development of a therapy that allows long-lived immune self-containment of HIV and restoration of these CTL responses by the host would be ideal. Through genetic manipulation of human blood-forming stem cells, we introduced a molecule– an HIV-targeting T cell receptor (TCR)–that allowed the generation of functional HIV-specific CTLs following differentiation within human tissues in a humanized mouse model. To assess if these newly developed, HIV-specific CTLs can allow active suppression of HIV replication, we infected these mice with HIV. We found that the development of genetically modified, HIV-specific CTLs in these mice results in the presence of a functional antiviral CTL response in vivo that significantly lowers viral replication following HIV infection. These results have strong implications for the use of this technology to engineer the human immune response to combat viral infections and suggest that genetic engineering via HSCs may allow tailoring of the immune response to target and eradicate HIV.
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影响因子：
3.7
作者：
Kitchen SG;Bennett M;Galić Z;Kim J;Xu Q;Young A;Lieberman A;Joseph A;Goldstein H;Ng H;Yang O;Zack JA
通讯作者：
Zack JA
影响因子：
9.8
作者：
Asquith B;Edwards CT;Lipsitch M;McLean AR
通讯作者：
McLean AR
影响因子：
4.2
作者：
Hughes, MS;Yu, YYL;Morgan, RA
通讯作者：
Morgan, RA
影响因子：
5.4
作者：
Brainard, Diana M.;Seung, Edward;Tager, Andrew M.
通讯作者：
Tager, Andrew M.
影响因子：
15.3
作者：
Appay, V;Nixon, D F;Donahoe, S M;Gillespie, G M;Dong, T;King, A;Ogg, G S;Spiegel, H M;Conlon, C;Spina, C A;Havlir, D V;Richman, D D;Waters, A;Easterbrook, P;McMichael, A J;Rowland-Jones, S L
通讯作者：
Rowland-Jones, S L