RevM10-expressing T cells derived in vivo from transduced human hematopoietic stem-progenitor cells inhibit human immunodeficiency virus replication
RevM10-expressing T cells derived in vivo from transduced human hematopoietic stem-progenitor cells inhibit human immunodeficiency virus replication
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DOI：
10.1128/jvi.71.6.4707-4716.1997
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发表时间：
1997-06-01
影响因子：
5.4
通讯作者：
Kaneshima, H
中科院分区：
文献类型：
--
作者：
Bonyhadi, ML;Moss, K;Kaneshima, H
人类免疫缺陷病毒1型(HIV-1)感染发病机制的一个关键特征是CD3+T细胞的逐渐丧失。为了抑制HIV在成熟T细胞中的复制，已经设计了一些基因治疗策略。由于T细胞是血液淋巴分化的产物，将抗病毒基因植入造血干细胞可以作为一种载体，为从转导干细胞衍生的后代T细胞提供长期保护。一种这样的“细胞免疫”策略利用了HIV-1rev反式显性突变蛋白RevM10的基因编码，该突变蛋白已被证明能抑制HIV-1在T细胞系和原代T细胞中的复制。在这项研究中，我们使用了基于Moloney小鼠白血病病毒的逆转录病毒，编码了同时表达RevM10和小鼠CD8-α‘链(Lyt2)的双顺反子信息。这种载体可以快速选择转基因表达细胞，并对转基因表达进行定量。我们证明了从人脐血或粒细胞集落刺激因子动员的外周血中分离的RevM10-CD34富集性造血祖干细胞(HPSC)可以在SCID-Hu胸腺/肝脏小鼠模型中分化为成熟的胸腺细胞。HPSC来源的胸腺细胞的表型分布是正常的，通过流式细胞仪分析可以检测到转基因的表达。此外，我们还证明了RevM10可以抑制体外扩增后转导的HPSC来源的T细胞中的HIV复制。这是第一次证明从转导的人HPSC中获得的T细胞具有抗HIV的功效。
A key feature of the pathogenesis of human immunodeficiency virus type 1 (HIV-1) infection is the gradual loss of CD3-positive T cells. A number of gene therapy strategies have been designed with the intent of inhibiting HIV replication in mature T cells. As T cells are products of hematolymphoid differentiation, insertion of antiviral genes into hematopoietic stem cells could serve as a vehicle to confer long-term protection in progeny T cells derived from transduced stem cells. One such ''cellular immunization'' strategy utilizes the gene coding for the HIV-1 rev trans-dominant mutant protein RevM10 which has been demonstrated to inhibit HIV-1 replication in T-cell lines and in primary T cells. In this study, we used a Moloney murine leukemia virus-based retrovirus encoding a bicistronic message coexpressing RevM10 and the murine CD8-alpha' chain (Lyt2). This vector allows rapid selection of transgene-expressing cells as well as quantitation of transgene expression. We demonstrate that RevM10-transduced CD34-enriched hematopoietic progenitor-stem cells (HPSC) isolated from human umbilical cord blood or from granulocyte colony-stimulating factor-mobilized peripheral blood can give rise to mature thymocytes in the SCID-hu thymus/liver mouse model. The phenotypic distribution of HPSC-derived thymocytes is normal, and expression of the transgene can be detected by how cytometric analysis. Moreover, we demonstrate that RevM10 can inhibit HIV replication in T cells derived from transduced HPSC after expansion in vitro. This is the first demonstration of anti-HIV efficacy in T cells derived from transduced human HPSC.