Ding C, Kume A, Björgvinsdóttir H, Hawley R G, Pech N, Dinauer M C
Herman B Wells Center for Pediatric Research, Department of Pediatrics (Hematology/Oncology), James Whitcomp Riley Hospital for Children, Indiana University Medical Center, Indianapolis, USA.
Blood. 1996 Sep 1;88(5):1834-40.
The X-linked form of chronic granulomatous disease (X-CGD) results from mutations in the gene encoding gp91phox, a 91-kD membrane glycoprotein that is the larger subunit of the respiratory burst oxidase cytochrome b. In this study, a new retroviral vector for expression of human gp91phox, MSCV-h91Neo, based on murine stem cell virus vectors, was evaluated using a human X-CGD myeloid cell line (X-CGD PLB-985 cells) and murine bone marrow cells. Expression of recombinant gp91phox in transduced X-CGD PLB-985 cells was substantially improved compared with levels achieved previously using a different retroviral construct, and respiratory burst oxidase activity was fully reconstituted in the majority of clones analyzed. Expression of gp91phox transcripts was also observed in primary and secondary murine colony-forming unit-spleen derived from transduced bone marrow cells. Furthermore, respiratory burst activity was restored to granulocyte-monocyte progeny of transduced X-CGD mice bone marrow cells cultured in vitro. This observation is the first reported use of gene transfer to correct the enzymatic defect in murine CGD phagocytes and is also consistent with the high conservation of the oxidase complex among different species. Taken together, these data suggest that the MSCV-h91Neo vector may be useful for gene replacement therapy in X-linked CGD, in which high-level reconstitution of phagocyte oxidase activity may be important for full correction of phagocyte microbicidal function.
X连锁慢性肉芽肿病（X-CGD）是由编码gp91phox的基因突变引起的，gp91phox是一种91-kD的膜糖蛋白，是呼吸爆发氧化酶细胞色素b的较大亚基。在本研究中，使用人X-CGD髓系细胞系（X-CGD PLB-985细胞）和小鼠骨髓细胞对基于小鼠干细胞病毒载体的用于表达人gp91phox的新型逆转录病毒载体MSCV-h91Neo进行了评估。与先前使用不同逆转录病毒构建体所达到的水平相比，转导的X-CGD PLB-985细胞中重组gp91phox的表达有显著改善，并且在大多数分析的克隆中呼吸爆发氧化酶活性完全重建。在转导的骨髓细胞衍生的原代和二代小鼠脾集落形成单位中也观察到了gp91phox转录本的表达。此外，转导的X-CGD小鼠骨髓细胞体外培养的粒细胞-单核细胞后代的呼吸爆发活性得以恢复。这一观察结果是首次报道使用基因转移来纠正小鼠CGD吞噬细胞中的酶缺陷，并且也与不同物种间氧化酶复合物的高度保守性一致。综上所述，这些数据表明MSCV-h91Neo载体可能对X连锁CGD的基因替代治疗有用，其中吞噬细胞氧化酶活性的高水平重建对于完全纠正吞噬细胞的杀菌功能可能很重要。
