成纤维细胞静脉注射途径实施血友病B基因治疗的初步探索

A preliminary study of intravenous injection of fibroblasts with gene transfer for clotting factorⅨminigene

将转有报告基因ｌａｃＺ的成纤维细胞３Ｔ３／ＢＡＧ以５×１０５量尾静脉注射到Ｂａｌｂ／ｃ小鼠体内，７ｄ后在主要脏器检测到３Ｔ３／ＢＡＧ的表达旦至少可持续３０ｄ．将转有人凝血因子Ⅸ小基因（ｈＦⅨｍｉｎｉｇｅｎｅ）的正向表达载体Ｇ１ＮａＣｉ＇和反向表达载体Ｇ１ＮａＣｉ＇ⅨＲ的成纤维细胞ＰＡ３１７／Ｇ１ＮａＣｉ＇Ⅸ和ＰＡ３１７／ＧＩＮａＣｉ＇ⅨＲ分别以１×１０＇量尾静脉注射到Ｂａｌｂ／ｃ小鼠体内，ＥＬＩＳＡ测定小鼠血浆中ｈＦⅨ蛋白的表达量，１６ｄ后，注射ＰＡ３１７／Ｇ１ＮａＣｉ＇Ⅸ的小鼠表达为（８．８９±０．５８）ｎｇ／ｍｌ，注射ＰＡ３１７／ＧＩＮａＣｉ＇ⅨＲ的小鼠的表达为（６９７±１９８）ｎｇ／ｍｌ．而只注射ＰＡ３１７细胞的小鼠表达一直维持在基底水平．

T3/BAG cells were intravenously injected into Balb/c mice via caudal vein at 5×10s cells per injection. The expression of reporter gene lacZ was detected in major organs on the 7th day after injection. The expression lasted at least for 30 days in all the . tested materials. Both PA317/GINaCi' Ⅳ and PA317/GINaCi' Ⅳ R cells were intravenously injected into Balb/c mice at 1×105 cells per injection and hF Ⅳ protein in mouse plasma was detected by ELISA. The expression level of PA317/GINaCi' Ⅳreached (8. 89±0. 58)ng/ml and PA317/GINaCi' Ⅳ R was (6. 97± 1. 98) ng/ml on the 16th day after injection. However, in the control that injected PA317 cells expression level of hF Ⅳ gene was not detectable in mice plasma.