He Huan, Xue Jing, Wang Weiming, Liu Lihong, Ye Chaobaihui, Cong Zhe, Kimata Jason T, Qin Chuan, Zhou Paul
1 The Unit of Anti-Viral Immunity and Genetic Therapy, the Key Laboratory of Molecular Virology and Immunology, the Institut Pasteur of Shanghai , Chinese Academy of Sciences, Shanghai, China .
2 Key Laboratory of Human Disease Comparative Medicine, Chinese Ministry of Health, Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Institute of Laboratory Animal Science , Chinese Academy of Medical Sciences and Comparative Medicine Center, Peking Union Medical College, Beijing, China .
Hum Gene Ther. 2017 Mar;28(3):271-285. doi: 10.1089/hum.2016.135. Epub 2016 Dec 29.
Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors efficiently transduce genes to human, but not rhesus, primary T cells and hematopoietic stem cells (HSCs). The poor transduction of HIV-1 vectors to rhesus cells is mainly due to species-specific restriction factors such as rhesus TRIM5α. Previously, several strategies to modify HIV-1 vectors were developed to overcome rhesus TRIM5α restriction. While the modified HIV-1 vectors efficiently transduce rhesus HSCs, they remain suboptimal for rhesus primary T cells. Recently, HIV-1 variants that encode combinations of LNEIE mutations in capsid (CA) protein and SIVmac239 Vif were found to replicate efficiently in rhesus primary T cells. Thus, the present study tested whether HIV-1 vectors packaged by a packaging construct containing these CA substitutions could efficiently transduce both human and rhesus primary CD4 T cells. To accomplish this, LNEIE mutations were made in the packaging construct CEMΔ8.9, and recombinant HIV-1 vectors packaged by Δ8.9 WT or Δ8.9 LNEIE were generated. Transduction rates, CA stability, and vector integration in CEMss-CCR5 and CEMss-CCR5-rhTRIM5α/green fluorescent protein cells, as well as transduction rates in human and rhesus primary CD4 T cells by Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors, were compared. Finally, the influence of rhesus TRIM5α variations in transduction rates to primary CD4 T cells from a cohort of 37 Chinese rhesus macaques was studied. While it maintains efficient transduction for human T-cell line and primary CD4 T cells, Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α-mediated CA degradation, resulting in significantly higher transduction efficiency of rhesus primary CD4 T cells than Δ8.9 WT-packaged HIV-1 vector. Rhesus TRIM5α variations strongly influence transduction efficiency of rhesus primary CD4 T cells by both Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors. Thus, it is concluded that Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α restriction and efficiently transduces both human and rhesus primary T cells.
基于1型人类免疫缺陷病毒（HIV-1）的慢病毒载体能有效地将基因转导至人类原代T细胞和造血干细胞（HSC），但不能转导至恒河猴原代T细胞。HIV-1载体对恒河猴细胞的转导效率低主要是由于物种特异性限制因子，如恒河猴TRIM5α。此前，已开发出几种修饰HIV-1载体的策略来克服恒河猴TRIM5α的限制。虽然修饰后的HIV-1载体能有效地转导恒河猴HSC，但对恒河猴原代T细胞来说仍不是最理想的。最近，发现编码衣壳（CA）蛋白中LNEIE突变组合和猴免疫缺陷病毒239（SIVmac239）Vif的HIV-1变体在恒河猴原代T细胞中能高效复制。因此，本研究测试了由含有这些CA替代的包装构建体包装的HIV-1载体是否能有效地转导人类和恒河猴原代CD4 T细胞。为实现这一目的，在包装构建体CEMΔ8.9中引入LNEIE突变，并产生由Δ8.9 WT或Δ8.9 LNEIE包装的重组HIV-1载体。比较了在CEMss-CCR5和CEMss-CCR5-rhTRIM5α/绿色荧光蛋白细胞中的转导率、CA稳定性和载体整合情况，以及由Δ8.9 WT或Δ8.9 LNEIE包装的HIV-1载体对人类和恒河猴原代CD4 T细胞的转导率。最后，研究了恒河猴TRIM5α变异对来自37只中国恒河猴群体的原代CD4 T细胞转导率的影响。虽然由Δ8.9 LNEIE包装的HIV-1载体对人类T细胞系和原代CD4 T细胞保持高效转导，但它克服了恒河猴TRIM5α介导的CA降解，导致其对恒河猴原代CD4 T细胞的转导效率显著高于由Δ8.9 WT包装的HIV-1载体。恒河猴TRIM5α变异对由Δ8.9 WT或Δ8.9 LNEIE包装的HIV-1载体转导恒河猴原代CD4 T细胞的效率有强烈影响。因此，得出结论：由Δ8.9 LNEIE包装的HIV-1载体克服了恒河猴TRIM5α的限制，能有效地转导人类和恒河猴原代T细胞。
