%0 Journal Article
%T A study of gene transfer and expression of human clotting factor IX in Hemophilia B mice mediated by mini-adenoviral vector
%A GAO Xiaobo
%A YE Chenbo
%A SHI Ding
%A CHEN Li
%A QIU Xinfang
%A XUE Jinglun
%A <br>高啸波
%A 叶晨波
%A 侍鼎
%A 陈立
%A 邱信芳
%A 薛京伦
%J 中国科学C辑(英文版)
%D 2003
%I Springer
%X Vector Gti'IX containing human clotting factor IX cDNAwith intron 1 (hFIX mini-gene or Fi'IX) driven by CMV promoter was constructed based on the mini-adenoviral vector GT2073 (mini-Ad vector) with all viral protein coding sequences deleted. Mini-Ad packaging cell 293Cre4 was first transduced with GTi'IX, and then was transfected with helper-adenovirus AdLC8, thus mini-Ad virions AdGTi'IX were obtained. At the same time, previous normal adenoviral vector pAdSPi'IX containing viral genome and hFIX mini-gene was constructed, and then previous adenovirus (pre-Ad) AdSPi'IX was obtained as control. The ratio of helper-adenovirus among purified virons AdGTi'IX was less than 0.8%. 3T3 cells were transfected with AdGTi'IX and AdSPi'IX at a MOI of 50 per cell and ELISA result showed that transient expression level in vitro was 1.4 +/-0.2 mug /10(6) . 24 h and 1.6 +/-0.3 mug/10(6) . 24 h respectively. Each hemophilia B (FIX knock-out) mouse received celiac injection of 1x10(10) pfu AdGTi'IX or AdSPi'IX. The highest expression level of hFIX in mouse plasma was 590 ng/mL and 690 ng/mL respectively, and the expression time lasted for 16 weeks and 9 weeks respectively. The bleeding time reduced from over 30 min to 7.5 min, and 5-min blood lost reduced from 430 muL to 60 muL. The results of anti-Ad IgG assays indicated that immune response triggered by AdGTi'IX was obviously weaker than that triggered by AdSPi'IX. These results indicated that, compared with previous adenovirus (pre-Ad), the mini-Ad vector system prolonged the expression time of hFIX and reduced immune response, thus offering a promising result for further pre-clinical study.
%K hemophilia B
%K mini-adenovirus
%K human clotting factor IX mini-gene
%K gene therapy
%K hemophilia B mouse<br>
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=180CF3A72E750F3261A8A60EDC957784&aid=79AEAE51CBEA1F086539A0BB94E7AFC4&yid=D43C4A19B2EE3C0A&vid=D997634CFE9B6321&iid=B31275AF3241DB2D&sid=250DF325A002B9CC&eid=DB7B2C790D19BE6E&journal_id=1674-7305&journal_name=ScienceChina.Lifesciences&referenced_num=0&reference_num=0