Molecular and Clinical Hematology Branch, National Heart Lung and Blood Institute-National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
Exp Hematol. 2013 Sep;41(9):779-788.e1. doi: 10.1016/j.exphem.2013.04.014. Epub 2013 May 9.
Innate immune factors, such as TRIM5α and cyclophilin A (CypA), act as a major restriction factor of retroviral infection among species. When HIV1 infects human cells, HIV1 capsid binds to human CypA to escape from human TRIM5α restriction. However, in rhesus cells, the mismatch between HIV1 capsid and rhesus CypA is recognized by rhesus TRIM5α to reduce HIV1 infectivity through proteasomal degradation. To circumvent this block, we previously developed a chimeric HIV1 vector (χHIV) that substituted HIV1 capsid with SIV capsid, and it significantly increased transduction efficiency for nonhuman primate cells. In this study, we evaluated whether the χHIV vector efficiently transduces human cells, and the transduction efficiency might increase by a CypA inhibitor (cyclosporine) and a proteasome inhibitor (MG132). The χHIV vector could transduce human CD34⁺ cells, as efficiently as the HIV1 vector, in vitro and in xenograft mice, even in the mismatch between SIV capsid and human CypA. Cyclosporine decreased transduction efficiency with the HIV1 vector, whereas it slightly increased transduction efficiency with the χHIV vector in human CD34⁺ cells. MG132 increased transduction efficiency with both χHIV and HIV1 vectors in the same manner. However, MG132 was toxic to human CD34⁺ cells at high concentrations, and both drugs had a small range of effective dosage. These findings demonstrate that both χHIV and HIV1 vectors have similar transduction efficiency for human hematopoietic repopulating cells, suggesting that the χHIV vector escapes from TRIM5α restriction, which is independent of human CypA.
先天免疫因子,如 TRIM5α 和亲环素 A(CypA),是物种间逆转录病毒感染的主要限制因子。当 HIV1 感染人类细胞时,HIV1 衣壳与人类 CypA 结合以逃避人类 TRIM5α 的限制。然而,在恒河猴细胞中,HIV1 衣壳与恒河猴 CypA 之间的不匹配被恒河猴 TRIM5α 识别,通过蛋白酶体降解降低 HIV1 的感染性。为了规避这一障碍,我们之前开发了一种嵌合 HIV1 载体(χHIV),用 SIV 衣壳替代了 HIV1 衣壳,它显著提高了非人类灵长类细胞的转导效率。在这项研究中,我们评估了 χHIV 载体是否能有效地转导人类细胞,以及 CypA 抑制剂(环孢素)和蛋白酶体抑制剂(MG132)是否能提高转导效率。χHIV 载体在体外和异种移植小鼠中,能够像 HIV1 载体一样有效地转导人类 CD34⁺ 细胞,即使在 SIV 衣壳和人类 CypA 之间不匹配的情况下也是如此。环孢素降低了 HIV1 载体的转导效率,而对人类 CD34⁺ 细胞中的 χHIV 载体的转导效率略有提高。MG132 以相同的方式增加了 χHIV 和 HIV1 载体的转导效率。然而,MG132 在高浓度时对人类 CD34⁺ 细胞有毒,并且两种药物的有效剂量范围都很小。这些发现表明,χHIV 和 HIV1 载体对人类造血重编程细胞具有相似的转导效率,表明 χHIV 载体逃避了 TRIM5α 的限制,这与人类 CypA 无关。