Kung Sam K P, An Dong-Sung, Bonifacino Aylin, Metzger Mark E, Ringpis Gene-Errol, Mao Si-Hua, Chen Irvin S Y, Donahue Robert E
Department of Microbiology, Immunology & Molecular Genetics, and Medicine, UCLA AIDS Institute, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, California 90095, USA.
Mol Ther. 2003 Dec;8(6):981-91. doi: 10.1016/j.ymthe.2003.08.020.
Modeling human hematopoietic progenitor cell gene therapy in nonhuman primates allows long-term evaluation of safety, maintenance of gene expression, and potential immune response against transgene products. We transplanted autologous G-CSF/SCF-mobilized CD34+ cells transduced with lentiviral vectors expressing EGFP into myeloablated rhesus macaques. To date, more than 4 years posttransplantation, 0.5-8% EGFP expression is maintained in multiple cell lineages. The animals remain healthy with no evidence of hematopoietic abnormalities or malignancies. To assess immune functions, we actively immunized two of our transplanted animals with purified rEGFP proteins and CpG adjuvant and demonstrated stable levels of EGFP+ cell populations maintained for over 29 months despite four active immunizations. We did not detect a persistent anti-EGFP antibody response or anti-EGFP T cell response in these immunized animals. Immune response to an irrelevant antigen was normal. Taken together, our data provide formal support that transplantation of lentivirally transduced CD34+ progenitor cells in myeloablated rhesus macaques induces specific immunological tolerance toward a foreign transgene.
在非人灵长类动物中对人类造血祖细胞基因治疗进行建模，可对安全性、基因表达的维持以及针对转基因产物的潜在免疫反应进行长期评估。我们将用表达增强绿色荧光蛋白（EGFP）的慢病毒载体转导的自体粒细胞集落刺激因子（G-CSF）/干细胞因子（SCF）动员的CD34⁺细胞移植到经清髓处理的恒河猴体内。迄今为止，在移植后4年多的时间里，多个细胞谱系中维持着0.5% - 8%的EGFP表达。这些动物保持健康，没有造血异常或恶性肿瘤的迹象。为了评估免疫功能，我们用纯化的重组EGFP蛋白和CpG佐剂对两只移植动物进行了主动免疫，结果表明，尽管进行了四次主动免疫，EGFP⁺细胞群体的水平仍稳定维持了29个月以上。在这些免疫动物中，我们未检测到持续的抗EGFP抗体反应或抗EGFP T细胞反应。对无关抗原的免疫反应正常。综上所述，我们的数据为在经清髓处理的恒河猴中移植慢病毒转导的CD34⁺祖细胞可诱导对异源转基因产生特异性免疫耐受提供了正式支持。
