Kalomoiris Stefanos, Lawson Je'tai, Chen Rachel X, Bauer Gerhard, Nolta Jan A, Anderson Joseph S
University of California Davis, Department of Internal Medicine, Sacramento, CA 95817, USA.
Hum Gene Ther Methods. 2012 Dec;23(6):366-75. doi: 10.1089/hgtb.2012.142. Epub 2013 Jan 11.
As HIV continues to be a global public health problem with no effective vaccine available, new and innovative therapies, including HIV gene therapies, need to be developed. Due to low transduction efficiencies that lead to low in vivo gene marking, therapeutically relevant efficacy of HIV gene therapy has been difficult to achieve in a clinical setting. Methods to improve the transplantation of enriched populations of anti-HIV vector-transduced cells may greatly increase the in vivo efficacy of HIV gene therapies. Here we describe the development of preselective anti-HIV lentiviral vectors that allow for the purification of vector-transduced cells to achieve an enriched population of HIV-resistant cells. A selectable protein, human CD25, not normally found on CD34+ hematopoietic progenitor cells (HPCs), was incorporated into a triple combination anti-HIV lentiviral vector. Upon purification of cells transduced with the preselective anti-HIV vector, safety was demonstrated in CD34+ HPCs and in HPC-derived macrophages in vitro. Upon challenge with HIV-1, improved efficacy was observed in purified preselective anti-HIV vector-transduced macrophages compared to unpurified cells. These proof-of-concept results highlight the potential use of this method to improve HIV stem cell gene therapy for future clinical applications.
由于艾滋病病毒(HIV)仍然是一个全球性公共卫生问题且尚无有效的疫苗,因此需要开发新的创新性疗法,包括HIV基因疗法。由于转导效率低导致体内基因标记率低,HIV基因疗法在临床环境中难以实现具有治疗意义的疗效。提高抗HIV载体转导细胞富集群体移植的方法可能会大大提高HIV基因疗法的体内疗效。在此,我们描述了预选择性抗HIV慢病毒载体的开发,该载体可用于纯化载体转导细胞,以获得富集的HIV抗性细胞群体。一种通常不存在于CD34+造血祖细胞(HPC)上的可选择蛋白——人CD25,被整合到一种三联组合抗HIV慢病毒载体中。在用预选择性抗HIV载体转导的细胞纯化后,在CD34+HPC和体外HPC衍生的巨噬细胞中证明了安全性。在用HIV-1攻击后,与未纯化的细胞相比,在纯化的预选择性抗HIV载体转导的巨噬细胞中观察到了更高的疗效。这些概念验证结果突出了该方法在改善HIV干细胞基因疗法以供未来临床应用方面的潜在用途。