Nunoi H, Ishibashi F
Department of Pediatrics, Kumamoto University School of Medicine.
Rinsho Byori. 1999 Jul;47(7):658-64.
Chronic granulomatous disease (CGD) is an inherited immune deficiency caused by mutations in any of the following four phox genes encoding subunits of the superoxide generating phagocyte NADPH oxidase. It consists of membranous cytochrome b558 composed of gp91-phox and p22-phox, and four cytosolic components, p47-phox, p67-phox, rac p21 and p40-phox, which translocate to the membrane upon activation. In our group study, more than 220 CGD patients has been enrolled. The incidence of CGD patients was estimated as 1 out of 250,000 births. The expected life span of the CGD patients is 25 to 30 years old by the Kaplan Meier analysis. Comparing with the ratio of CGD subtype in US and Europe, that with p47-phox deficiency is lower (less than 10% vs. 23%) and that of gp91-phox deficiency is higher (more than 75% vs. 60%). Prophylactic administration of ST antibiotics and IFN-gamma and bone marrow transplantation have been successfully employed in our therapeutic strategy. However, it is necessary to develop the gene therapy technology for CGD patients as more promising treatment. In the current study we constructed two retrovirus vectors; MFGS-gp91/293 SPA which contains only the therapeutic gp91-phox gene, a bicistronic retrovims pHa-MDR-IRES-gp91/PA317 which carries a multi drug resistant gene (MDR1) and the gp91-phox gene connected with an internal ribosome entry site (IRES). We demonstrate high efficiency transduction of gp91-phox to CGD EB virus established cell line with high levels of functional correction of the oxidase by MFGS-gp91 and by pHa-MDR-IRES-gp91, respectively. We also demonstrate sufficient transduction of gp91-phox to CD34+ haematopoietic stem cell from the patients with gp91-phox deficiency by MFGS-gp91/293 SPA. Our current studies suggest that the combination of the 293-SPA packaging system and the bicistronic retrovirus system inserted MDR1 gene make our CGD gene therapy more feasible for clinical application.
慢性肉芽肿病（CGD）是一种遗传性免疫缺陷病，由编码超氧化物生成吞噬细胞NADPH氧化酶亚基的以下四个phox基因中的任何一个发生突变引起。它由gp91-phox和p22-phox组成的膜细胞色素b558以及四个胞质成分p47-phox、p67-phox、rac p21和p40-phox组成，这些成分在激活后会转移到膜上。在我们的小组研究中，已招募了220多名CGD患者。据估计，CGD患者的发病率为每25万例出生中有1例。通过Kaplan Meier分析，CGD患者的预期寿命为25至30岁。与美国和欧洲的CGD亚型比例相比，p47-phox缺陷型的比例较低（不到10%对23%），而gp91-phox缺陷型的比例较高（超过75%对60%）。预防性使用ST抗生素、干扰素-γ和骨髓移植已成功应用于我们的治疗策略中。然而，有必要为CGD患者开发更有前景的基因治疗技术。在当前研究中，我们构建了两种逆转录病毒载体；仅包含治疗性gp91-phox基因的MFGS-gp91/293 SPA，以及携带多药耐药基因（MDR1）和与内部核糖体进入位点（IRES）连接的gp91-phox基因的双顺反子逆转录病毒pHa-MDR-IRES-gp91/PA317。我们分别证明了MFGS-gp91和pHa-MDR-IRES-gp91对CGD EB病毒建立的细胞系高效转导gp91-phox，并对氧化酶进行了高水平的功能校正。我们还证明了MFGS-gp91/293 SPA对来自gp91-phox缺陷患者CD34+造血干细胞的gp91-phox进行了充分转导。我们目前的研究表明，293-SPA包装系统和插入MDR1基因的双顺反子逆转录病毒系统的组合使我们的CGD基因治疗在临床应用中更可行。
