Marking and gene expression by a lentivirus vector in transplanted human and nonhuman primate CD34+ cells
Marking and gene expression by a lentivirus vector in transplanted human and nonhuman primate CD34+ cells
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DOI：
10.1128/jvi.74.3.1286-1295.2000
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发表时间：
2000-02-01
影响因子：
5.4
通讯作者：
Donahue, RE
中科院分区：
文献类型：
--
作者：
An, DS;Wersto, RP;Donahue, RE
近来，基于人类免疫缺陷病毒（HIV）的基因传递载体已被开发出来，作为一种基因传递的替代方式。这些载体具有若干优势，特别是在感染非活跃分裂细胞的能力方面。然而，使用基于人类免疫缺陷病毒的载体引发了诸多问题，其中安全问题尤为重要；因此，在灵长类动物模型系统中对不同造血谱系的标记和基因表达进行进一步的特性研究是可取的。我们使用两种基因治疗的动物模型系统来测试转导和标记的效率，以及这些载体的安全性。第一种利用恒河猴动物模型进行细胞因子动员的自体外周血CD34(+)细胞移植。第二种使用严重联合免疫缺陷 - 人（SCID - hu）胸腺/肝脏嵌合移植动物模型，该模型对人类T淋巴细胞祖细胞的重建特别有用。在恒河猴中，在粒细胞、淋巴细胞、单核细胞中观察到可检测水平的载体，并且在一只标记水平最高的动物中，在红细胞和血小板中也观察到了载体。在移植的SCID - hu小鼠中，我们直接比较了慢病毒载体和鼠白血病病毒衍生载体在胸腺细胞中的标记和基因表达。观察到的标记水平相当，但携带巨细胞病毒内部启动子的慢病毒载体的表达效率低于由其自身长末端重复序列表达的鼠逆转录病毒载体。在对1型传染性人类免疫缺陷病毒（HIV - 1）的检测中，在两种动物模型系统中均未检测到具有复制能力的HIV - 1。因此，这些结果表明，虽然慢病毒载体没有明显的有害影响，并且可能比鼠逆转录病毒载体具有优势，但对其最佳使用的要求进行进一步研究是有必要的。
Recently, gene delivery vectors based on human immunodeficiency virus (HIV) have been developed as an alternative mode of gene delivery. These vectors have a number of advantages, particularly in regard to the ability to infect cells which are not actively dividing. However, the use of vectors based on human immunodeficiency virus raises a number of issues, not the least of which is safety; therefore, further characterization of marking and gene expression in different hematopoietic lineages in primate animal model systems is desirable. We use two animal model systems for gene therapy to test the efficiency of transduction and marking, as web as the safety of these vectors. The first utilizes the rhesus animal model for cytokine-mobilized autologous peripheral blood CD34(+) cell transplantation. The second uses the SCID-human (SCID-hu) thymus/liver chimeric graft animal model useful specifically for human T-lymphoid progenitor cell reconstitution. In the rhesus macaques, detectable levels of vector were observed in granulocytes, lymphocytes, monocytes, and, in one animal with the highest levels of marking, erythrocytes and platelets. In transplanted SCID-hu mice, we directly compared marking and gene expression of the lentivirus vector and a murine leukemia virus-derived vector in thymocytes. Marking was observed at comparable levels, but the lentivirus vector bearing an internal cytomegalovirus promoter expressed less efficiently than did the murine retroviral vector expressed from its own long terminal repeats. In assays for infectious HIV type 1 (HIV-1), no replication-competent HIV-1 was detected in either animal model system. Thus, these results indicate that while lentivirus vectors have no apparent deleterious effects and may have advantages over murine retroviral vectors, further study of the requirements for optimal use are warranted.