Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-1203, USA.
Mol Ther. 2011 Jan;19(1):113-21. doi: 10.1038/mt.2010.203. Epub 2010 Sep 21.
To identify cellular promoters in a self-inactivating (SIN) lentiviral vector that might be beneficial in treating children with leukocyte adhesion deficiency type 1 (LAD-1), we tested lentiviral vectors with human CD11 and CD18 leukocyte integrin proximal promoter elements directing expression of canine CD18 in animals with canine LAD (CLAD). Lentiviral vectors with either the human CD11b (637 bp) proximal promoter or the human CD18 (1,060 bp) proximal promoter resulted in the highest percentages of CD18(+) CLAD CD34(+) cells in vitro. Subsequently, two CLAD dogs were infused with autologous CD34(+) cells transduced with the hCD11b (637 bp)-cCD18 vector, and two CLAD dogs were infused with autologous CD34(+) cells transduced with the hCD18 (1,060 bp)-cCD18 vector. Each dog received a nonmyeloablative dose of 200 cGy total body irradiation (TBI) before the infusion of transduced cells. The two CLAD dogs treated with the hCD18 (1,060 bp)-cCD18 vector, and one of the two dogs treated with the hCD11b (637 bp)-cCD18 vector, had reversal of the CLAD phenotype. These studies using endogenous leukocyte integrin proximal promoters represent an important step in the development of gene therapy for children with LAD-1.
为了鉴定自我失活（SIN）慢病毒载体中的细胞启动子，这些启动子可能有益于治疗白细胞黏附缺陷 1 型（LAD-1）患儿，我们在患有犬类 LAD（CLAD）的动物中测试了带有犬类 CD18 表达的人 CD11 和 CD18 白细胞整合素近端启动子元件的慢病毒载体。带有人 CD11b（637bp）近端启动子或人 CD18（1,060bp）近端启动子的慢病毒载体在体外导致最高百分比的 CD18(+) CLAD CD34(+)细胞。随后，用 hCD11b(637bp)-cCD18 载体转导的自体 CD34(+)细胞输注给两只 CLAD 犬，用 hCD18(1,060bp)-cCD18 载体转导的自体 CD34(+)细胞输注给另外两只 CLAD 犬。在输注转导细胞之前，每只狗接受 200cGy 全身照射（TBI）的非清髓剂量。用 hCD18(1,060bp)-cCD18 载体治疗的两只 CLAD 犬和用 hCD11b(637bp)-cCD18 载体治疗的两只犬中的一只，CLAD 表型得到逆转。这些使用内源性白细胞整合素近端启动子的研究代表了为 LAD-1 患儿开发基因治疗的重要一步。
