Induction of transplantation tolerance based on bio-molecular technique

基于生物分子技术的移植耐受诱导

基本信息

批准号：
10307030
负责人：
SUZUKI Seiichi
金额：
$ 21.25万
依托单位：
National Children's Medical Research Center
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

An adenovirus vector AxCALNFasL was constructed in order to transduce a gene for Fas-Ligand(FasL), requiring cotransfection of Cre recombinase-expressing vector AxCANCre for its expression. When intravenously injected AxCALNFasL, rat liver expressed FasL by co-administration of AxCANCre in a dose-dependent manner. The FasL-expressed livers survived significantly in the allogeneic recipients. However, a high level of FasL-expression induced animal death due to fulminant hepatitis. Furthermore, we successfully generated AxCALNLCrmA, a recombinant adenovirus expressing CrmA-gene with a Cre-mediated switching cassette. When allografted, the CrmA-expressed livers were resistant to rejection response in the recipients, resulting in marked prolongation of recipient survival. The formation of active caspase-3 was markedly inhibited in the Crm A gene-transfected hepatocytes. In addition, CTLA4Ig, a soluble recombinant fusion protein that contains the extracellular domain of CTLA4 and Fc portion … More of IgG1, strongly adheres to the B7 to block CD28-mediated costimulatory signals and inhibits in vitro and in vivo immune responses. We intravenously injected the adenovirus vector containing CTLA4Ig cDNA(AxCAhCTLA4Ig)to the heartgrafted rats immediately after grafting. The serum level of CTLA4Ig reached to maximum at 51-93μg/ml 3 to 7 days after gene-transfection and declined after 14 days, although detectable levels were observed up to 49 days. The graft survival time was significantly prolonged in the gene-transfected recipient rats. Down-regulation of IL-2 and IFNγ mRNA and persistence of IL-4 and IL-10 transcripts were observed in the graft-infiltrating cells. Peri-operative administration of FTY720, a new immunosuppressant inducing lymphocyte apoptosis, in combination with the CTLA4Ig gene-transfection resulted in synergistic prolongation of graft survival, and 33% of recipients acquired transplantation tolerance. We detected CTLA4Ig protein in the sera 49 days after grafting ; its levels were always hiher in the combination group than in the AxCAhCTLA4Ig-alone group. Thus, FTY720 would be highly effective for enhancing the CTLA4Ig expression by adenovirus-mediated gene transfer. Less

构建腺病毒载体AxCALNFasL以转导Fas配体(FasL)基因，需要共转染Cre重组酶表达载体AxCANCre才能表达。当静脉注射AxCALNFasL时，通过与AxCANCre共施用，大鼠肝脏表达FasL。然而，FasL 表达的肝脏在同种异体受体中显着存活。高水平的FasL表达诱导动物因暴发性肝炎而死亡此外，我们成功地产生了AxCALNLCrmA，一种表达CrmA基因的重组腺病毒，具有Cre介导的转换盒，当同种异体移植时，表达CrmA的肝脏对排斥反应具有抵抗力。 Crm A 中活性 caspase-3 的形成受到显着抑制。此外，CTLA4Ig是一种可溶性强重组融合蛋白，含有CTLA4的胞外结构域和IgG1的Fc部分，粘附在B7上以阻断CD28介导的共刺激信号并抑制体外和体内免疫。移植心脏后，我们立即静脉注射含有CTLA4Ig cDNA（AxCAhCTLA4Ig）的腺病毒载体。移植后3至7天，CTLA4Ig的血清水平达到最高，为51-93μg/ml，并在14天后下降，尽管可检测到的水平长达49天。移植物的存活时间显着延长。 -在移植物浸润细胞中观察到转染的受体大鼠IL-2和IFNγmRNA的下调以及IL-4和IL-10转录物的持续存在。 FTY720（一种诱导淋巴细胞凋亡的新型免疫抑制剂）与 CTLA4Ig 基因转染相结合，可协同延长移植物存活时间，33% 的受者在移植后 49 天检测到血清中 CTLA4Ig 蛋白水平；组合组中的值总是高于 AxCAhCTLA4Ig 单独组中的值，因此，FTY720 的值会更高。通过腺病毒介导的基因转移有效增强 CTLA4Ig 表达。

项目成果

期刊论文数量（142）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A. Tamura, et al: "Combination effect of tacrolimus and FTY720 in liver transplantation in rats"Transplant Proc. 31(7). 2785-2786 (1999)

A. Tamura 等人：“他克莫司和 FTY720 在大鼠肝移植中的联合作用”Transplant Proc。

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Yoshihiko Kageyama et al: "Apoptosis is involved in acute cardiac allograft rejection in rats." Ann Thor Surg. 65・6. 1604-1609 (1998)

Yoshihiko Kageyama 等人：“细胞凋亡与大鼠的急性同种异体移植排斥有关。”Ann Thor Surg 65·6（1998）。

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Torayuki Okuyama et al: "Efficient Fas-ligand gene expression in rodent liver after intravenous injection of a recombinant adenovirus by the use of a Cre-mediated switching system." Gene Therapy. 5. 1047-1053 (1998)

Torayuki Okuyama 等人：“通过使用 Cre 介导的转换系统，静脉注射重组腺病毒后，在啮齿动物肝脏中实现 Fas 配体基因的高效表达。”

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Masayuki Fujino, Xiao-Kang Li, Yusuke Kitazawa, Naoko Funeshima, Lei Guo, Torayuki Okuyama, Takashi Amano, Hiroshi Amemiya, Seiichi Suzuki: "Selective repopulation of mice liver after fas-resistant hepatocytes transplantation."Cell Transplantation. (in pr

Masayuki Fujino、Xiao-Kang Li、Yusuke Kitazawa、Naoko Funeshima、Lei Guo、Torayuki Okuyama、Takashi Amano、Hiroshi Amemiya、Seiichi Suzuki：“fas 抗性肝细胞移植后小鼠肝脏的选择性再增殖。”细胞移植。

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Masakatsu Yamashita, Makoto Katsuyama, Makio Iwashima, Motoko Kimura, Chiori Shimizu, Tohru Kamata, Tahiro Shin, Nobuko Seki, Seiichi Suzuki, Masaru Taniguchi, Toshinori Nakayama.: "TCR-induced calcineurin activation regulates Th2 cell development by modi

Masakatsu Yamashita、Makoto Katsuyama、Makio Iwashima、Motoko Kimura、Chiori Shimizu、Tohru Kamata、Tahiro Shin、Nobuko Seki、Seiichi Suzuki、Masaru Taniguchi、Toshinori Nakayama。：“TCR 诱导的钙调磷酸酶激活通过 modi 调节 Th2 细胞发育”