Long-term engraftment of nonobese diabetic/severe combined immunodeficient mice with human CD34+ cells transduced by a self-inactivating human immunodeficiency virus type 1 vector.
Long-term engraftment of nonobese diabetic/severe combined immunodeficient mice with human CD34+ cells transduced by a self-inactivating human immunodeficiency virus type 1 vector.
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将由自失活人类免疫缺陷病毒 1 型载体转导的人类 CD34 细胞长期植入非肥胖糖尿病/严重联合免疫缺陷小鼠中。
DOI：
10.1089/104303401750214294
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Garcia,JV
中科院分区：
文献类型：
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作者：
Gatlin,J;Padgett,A;Melkus,MW;Kelly,PF;Garcia,JV
具有活体繁殖潜能的人造血细胞有望成为许多遗传性或获得性造血疾病的矫正基因转移的靶点。在这里，我们展示了由编码增强型绿色荧光蛋白(EGFP)的基于HIV-1的自失活(SIN)载体转导的人CD34+细胞对非肥胖型糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠的长期造血重建。在没有细胞因子预刺激的情况下，人脐血CD34+细胞在低复数(MOI为5)的感染下被转导(高达76%)。将转导的hCD34+细胞导入受者体内，移植后18周可获得多系植入和稳定的转基因表达。移植小鼠的骨髓含有高达50%的hCD45+细胞和高达63%的hCD45+/EGFP+细胞。髓外脾重建分析显示高达13%的hCD45+细胞和高达41%的hCD45+/EGFP+细胞。对从受体动物骨髓中分离的人类祖细胞的分析显示，荧光显微镜和前病毒序列的PCR分析显示，EGFP+集落形成细胞(CFCs)的比例相等，表明体内转基因沉默最少。这些发现证明了慢病毒为基础的SIN载体用于造血干细胞基因转移的有效性，并为其未来的临床评估提供了有力的支持。
Human hematopoietic cells within vivorepopulating potential hold much promise as a target for corrective gene transfer for numerous inherited or acquired hematopoietic disorders. Here we demonstrate long-term hematopoietic reconstitution of nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice with human CD34+cells transduced by an HIV-1-based self-inactivating (SIN) vector encoding the enhanced green fluorescent protein (EGFP). Human umbilical cord CD34+cells were transduced (up to 76%) at a low multiplicity of infection (MOI of 5) in the absence of cytokine prestimulation. Introduction of transduced hCD34+cells into irradiated recipients resulted in multilineage engraftment and stable transgene expression for 18 weeks posttransplantation. Bone marrow from transplanted mice contained up to 50% hCD45+cells and up to 63% hCD45+/EGFP+cells. Analysis of extramedullar splenic reconstitution showed up to 13% hCD45+cells and up to 41% hCD45+/EGFP+cells. Analysis of human progenitor cells isolated from bone marrow of recipient animals showed equivalent percentages of EGFP+colony-forming cells (CFCs) by fluorescence microscopy and by PCR analysis of provirus sequences, indicating minimal transgene silencingin vivo. These findings demonstrate the utility of lentivirus-based SIN vectors for hematopoietic stem cell gene transfer and provide strong support for their future clinical evaluation.