摘要
本文报道了皮肤成纤维细胞基因治疗血友病Ｂ的新西兰大白兔试验及安全性检测。转有人凝血因子ＩＸＣＤＮＡ的免皮肤成纤维细胞包埋于胶原基质中，然后进行不同方式的自体或同种异体移植，移植兔血浆中人ＩＸ因子蛋白有高水平的表达且持续稳定。腹腔移植有ＲＳＦ－ＸＬＩＸ转化细胞的兔血浆中人ＩＸ因子表达水平达１００ｎｇ／ｍｌ，持续５个月以上；腹腔移植有ＲＳＦ－ｐＣＭＶＩＸ转化细胞的兔血浆中人ＩＸ因子表达水平达２９５ｎｇ／ｍｌ，持续１５个月以上；皮下细胞胶原液注射移植ＲＳＦ－Ｎ２ＣＭＶＩＸ转化细胞的兔血浆中人 ＩＸ因子表达水平达 ４８０ｎｇ／ｍｌ，持续 １０个月以上。 Ｗｅｓｔｅｒｎ ｂｌｏｔ证明血浆中表达的的确是人ＩＸ因子蛋白；安全性检测证明转化细胞移植途径安全可靠，没有检测到野生型病毒和恶性转化细胞，无任何毒副作用。
This paper reports the New Zealand rabbit experiment of gene transfer study for hemophilia B with skin fibroblasts. Rabbit skin fibroblasts transduced with retroviral vector containing human factor IX cDNA, were embedded in collagen and implanted into New Zealand rabbit autograftly or allograftly. Human FIX in plasma of rabbits intraperitoneally implanted with RSF-XLIX cells reached to 100ng/ml and has been lasting for over 5.5 months; Human FIX in rabbits intraperitoneally implanted with RSF-pCMVIX transformed cells reached to 295 ng/ml and has been lasting for 2 years;Human FIX in plasma of rabbits subcutaneously injected with cell collagen matrix of RSF-N2CMVIX reaced to 480ng/ml and has been lasting over 500 days. Western blot confirmed the 55KD protein expresed in rabbit plasma was indeed human FIX. Safty assesements showed that the approach of cell implantation was safe and no replication-competient virus, malignant transfomed cells & deterious effects were detected.
出处
《动物学报》
SCIE
CAS
CSCD
1994年第2期182-190,共9页
ACTA ZOOLOGICA SINICA
基金
国家"863"高技术发展计划资助项目863-102-17-(2)。
关键词
基因治疗
血友病
动物试验
Gene therapy, Gene transfer, Implantation of cell-collagen, Human factor IX, Safty assesement.
