Li X, Mukai T, Young D, Frankel S, Law P, Wong-Staal F
Department of Medicine, University of California, San Diego, La Jolla, USA.
J Hum Virol. 1998 Jul-Aug;1(5):346-52.
To use HIV-1 vectors to mediate stable gene transfer into hematopoietic stem/progenitor cells.
STUDY DESIGN/METHODS: Purified human CD34+ cells were transduced with HIV-1 vectors pseudotyped with VSV-G and subjected to colony-forming assays and differentiation in liquid culture. Transduction was determined by DNA-polymerase chain reaction (PCR) for the transgene. GFP reporter gene expression and phenotypes of progeny cells were assessed by microscopy and flow cytometry.
The HIV-1 vector transduced CD34+ cells with high efficiency. Transduction did not interfere with CD34+ cells differentiation in vitro. Transduced genes are expressed in different subsets of progeny cells, including those with normal dendritic cells (DC) morphology and phenotypes (HLADR+/CD1a+/CD86+/CD14-).
We have demonstrated efficient transduction of hematopoietic progenitor cells by HIV-1 vectors. The transgenes are expressed in different subsets of progeny cells, which suggests stable integration. The generation of DCs stably expressing HIV antigens provides a new approach for vaccine development.
利用HIV-1载体介导基因稳定转入造血干/祖细胞。
研究设计/方法：用VSV-G假型化的HIV-1载体转导纯化的人CD34+细胞，并进行集落形成试验及液体培养分化。通过针对转基因的DNA聚合酶链反应（PCR）测定转导情况。通过显微镜检查和流式细胞术评估GFP报告基因表达及子代细胞的表型。
HIV-1载体高效转导CD34+细胞。转导不干扰CD34+细胞的体外分化。转导基因在子代细胞的不同亚群中表达，包括那些具有正常树突状细胞（DC）形态和表型（HLADR+/CD1a+/CD86+/CD14-）的细胞。
我们已证明HIV-1载体可有效转导造血祖细胞。转导基因在子代细胞的不同亚群中表达，这表明整合稳定。稳定表达HIV抗原的DC的产生为疫苗开发提供了一种新方法。
