The induction of tolerance using the brand new gene therapy in transplantation setting and clarification of the mechanism.

在移植环境中使用全新基因疗法诱导耐受并阐明其机制。

基本信息

批准号：
13470248
负责人：
FURUKAWA Hiroyuki
金额：
$ 9.15万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

1. To develop innovative strategies for transplant immunosuppression, we tested the effect of the blockade of costimulatory pathways by using adenovirus vector coding CTLA4Ig(AdCTLA4Ig) or CD40Ig(AdCD40Ig)genes in ACI to LEW rat organ transplantations.A) Heart transplantation : A single treatment with AdCTLA4Ig or AdCD40Ig prolonged graft survival. Although the combined gene therapy allowed long term graft survival, this strategy was not enough to induce tolerance and to avoid chronic rejection.B) Liver transplantation : A single treatment with AdCTLA4Ig or AdCD40Ig prolonged graft survival, and could induce donor-specific tolerance.C) Small bowel transplantation : Although the combined gene therapy prolonged graft survival, this strategy was not enough to induce tolerance and to avoid chronic rejection.D) Xeno heart transplantation(Hamster to Rat) : DXR and cellular rejection were controlled successfully and all xenografts were accepted for over 100 days when AdCTLA4Ig and AdCD40Ig we … More re administered under FK779 induction therapy. However, chronic rejection was inevitable in the long-term surviving xenografts.2. Several strategies have been made in order to avoid the toxicity of adenoviral vector and to establish an efficient and safety gene transfer.A) Ex-vivo transfer : In rat liver transplantation, the graft with 1x10^9pfu AdCTLA4Ig in 4℃ UW solution for more than 8hrs could get efficient transfection rate and tolerance.B) Modified Clamp Technique : Using the short term CT transduction method (under the condition of 4℃ cold preservation for 10 minutes) long term graft survival are achieved.C) Cre/loxP : The administration of adenovirus vector containing Cre recombinase gene(AdexCACre) at the desired time induced Cre-mediated recombination within a gene derived from Adex1CALoxCTLA4IgGLox vector, and the cDNA of CTLA4IgG was excised from the transduced gene and terminated the expression of CTLA4IgG in vitro and in vivo.3. Immunosuppressive mechanism of blockade of CD40-CD 154A) CD40Ig has bi-directional effects, i.e., one direction towards CD4 + T cells that enhances IL-2 production by cross-linking CD40L, and the other to APCs, which results in generation of CD4 + CD25 + T cells. These events, in concert, lead to generation of functional CD4 + CD25 + T cells that regulate CTL activity in an antigen-specific manner. Less

1。开发用于移植的免疫选择的创新策略IRUS矢量编码CTLA4IG（ADCTLA4IG）或CD40ig（ADCD40IG）基因在ACI中以升级大鼠器官移植b）肝移植：用adctla4ig或adcd40ig延长移植物的单一治疗，并且可以特异性的耐受性。 .d）Xeno心脏移植（仓鼠对大鼠）：DXR和细胞排斥的控制在100天内被接受，而ADCTLA4IG和ADCD40ig我们在FK779指示治疗中不可避免地拒绝。异种移植。2。 b）修饰的夹具技术：在4°C冷保留10分钟下）ERM移植物存活率。从ctla4igg的Adex1caloxctla4igglox载体中衍生的基因中的募集，从ctla4igg的转导的表达中切除了体外和体内的ctla4igg的表达。3迈向增强IL-2 D40L的CD4 + T细胞，另一个会导致CD4 + CD25 + T细胞的产生，从而导致功能性CD4 + CD25 + T细胞的产生抗原特异性的活动。