Postentry restriction to human immunodeficiency virus-based vector transduction in human monocytes
Postentry restriction to human immunodeficiency virus-based vector transduction in human monocytes
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DOI：
10.1128/jvi.75.12.5448-5456.2001
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发表时间：
2001-06-01
影响因子：
5.4
通讯作者：
Collins, M
中科院分区：
文献类型：
--
作者：
Neil, S;Martin, F;Collins, M
单核细胞谱系的细胞可以在临床感染期间和体外被人免疫缺陷病毒1型（HIV-1）感染。因此，检查了基于HIV-1的载体活化人单核细胞、单核细胞衍生的巨噬细胞和树突细胞（DC）的能力，以开发将抗原基因递送至人抗原呈递细胞的有效方案。新鲜分离的单核细胞对基于HIV-1的载体转导难治，但在体外分化为成熟巨噬细胞后变得可转导。这种成熟依赖性转导不依赖于包装细胞中的HIV-1辅助蛋白Vif、Vpr、Vpu和Nef以及载体中的中央多嘌呤段，并且还观察到水泡性口炎病毒假型HIV-1前病毒，其仅在包膜和Nef中有缺陷，HIV-1为基础的载体的逆转录的水平和程度是相似的感染后，未成熟的单核细胞和成熟的巨噬细胞。然而，在单核细胞中不能检测到2LTR载体环，表明这些细胞中载体核进入的阻断。暴露于基于HIV-1的载体的新鲜分离的单核细胞的转导可以通过随后向DC的分化而被拯救，这种拯救是由DC培养基中的胎牛血清诱导的，其促进载体核进入。
Cells of the monocyte lineage can be infected with human immunodeficiency virus type 1 (HIV-1) both during clinical infection and in vitro. The ability of HIV-l-based vectors to transduce human monocytes, monocyte-derived macrophages, and dendritic cells (DCs) was therefore examined, in order to develop an efficient protocol for antigen gene delivery to human antigen-presenting cells. Freshly isolated monocytes were refractory to HIV-1-based vector transduction but became transducible after in vitro differentiation to mature macrophages. This maturation-dependent transduction was independent of the HIV-1 accessory proteins Vif, Vpr, Vpu, and Nef in the packaging cells and of the central polypurine tract in the vector, and it was also observed with a vesicular stomatitis virus-pseudotyped HIV-1 provirus, defective only in envelope and Nef, The level and extent of reverse transcription of the HIV-1-based vector was similar after infection of immature monocytes and of mature macrophages. However, 2LTR vector circles could not be detected in monocytes, suggesting a block to vector nuclear entry in these cells. Transduction of freshly isolated monocytes exposed to HIV-1-based vector could be rescued by subsequent differentiation into DCs, This rescue was induced by fetal calf serum in the DC culture medium, which promoted vector nuclear entry.