Sakuraba Hitoshi, Murata-Ohsawa Mai, Kawashima Ikuo, Tajima Youichi, Kotani Masaharu, Ohshima Toshio, Chiba Yasunori, Takashiba Minako, Jigami Yoshifumi, Fukushige Tomoko, Kanzaki Tamotsu, Itoh Kohji
Department of Clinical Genetics, The Tokyo Metropolitan Institute of Medical Science, Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo, 113-8613, Japan.
CREST, JST, Kawaguchi, Japan.
J Hum Genet. 2006;51(3):180-188. doi: 10.1007/s10038-005-0342-9. Epub 2005 Dec 22.
We compared two recombinant alpha-galactosidases developed for enzyme replacement therapy for Fabry disease, agalsidase alfa and agalsidase beta, as to specific alpha-galactosidase activity, stability in plasma, mannose 6-phosphate (M6P) residue content, and effects on cultured human Fabry fibroblasts and Fabry mice. The specific enzyme activities of agalsidase alfa and agalsidase beta were 1.70 and 3.24 mmol h(-1) mg protein(-1), respectively, and there was no difference in stability in plasma between them. The M6P content of agalsidase beta (3.6 mol/mol protein) was higher than that of agalsidase alfa (1.3 mol/mol protein). The administration of both enzymes resulted in marked increases in alpha-galactosidase activity in cultured human Fabry fibroblasts, and Fabry mouse kidneys, heart, spleen and liver. However, the increase in enzyme activity in cultured fibroblasts, kidneys, heart and spleen was higher when agalsidase beta was used. An immunocytochemical analysis revealed that the incorporated recombinant enzyme degraded the globotriaosyl ceramide accumulated in cultured Fabry fibroblasts in a dose-dependent manner, with the effect being maintained for at least 7 days. Repeated administration of agalsidase beta apparently decreased the number of accumulated lamellar inclusion bodies in renal tubular cells of Fabry mice.
我们比较了两种为法布里病酶替代疗法开发的重组α-半乳糖苷酶，即α-半乳糖苷酶A和β-半乳糖苷酶，比较内容包括特定α-半乳糖苷酶活性、在血浆中的稳定性、甘露糖6-磷酸（M6P）残基含量，以及对培养的人法布里成纤维细胞和法布里病小鼠的影响。α-半乳糖苷酶A和β-半乳糖苷酶的比酶活性分别为1.70和3.24 mmol h(-1) mg蛋白(-1)，它们在血浆中的稳定性没有差异。β-半乳糖苷酶的M6P含量（3.6 mol/mol蛋白）高于α-半乳糖苷酶A（1.3 mol/mol蛋白）。两种酶的给药均导致培养的人法布里成纤维细胞以及法布里病小鼠的肾脏、心脏、脾脏和肝脏中的α-半乳糖苷酶活性显著增加。然而，使用β-半乳糖苷酶时，培养的成纤维细胞、肾脏、心脏和脾脏中的酶活性增加更高。免疫细胞化学分析显示，掺入的重组酶以剂量依赖性方式降解培养的法布里成纤维细胞中积累的球三糖神经酰胺，且该效应至少维持7天。重复给予β-半乳糖苷酶明显减少了法布里病小鼠肾小管细胞中积累的层状包涵体数量。
