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Regression of established murine adenocarcinoma by in vivo allogeneic MHC gene transfer using electroporation and modification of donor MHC antigens by antisense gene transfer to prevent rejection of transplanted organs

使用电穿孔进行体内同种异体 MHC 基因转移并通过反义基因转移修饰供体 MHC 抗原以防止移植器官排斥，从而消退已形成的小鼠腺癌

基本信息

批准号：
10671157
负责人：
SHIMIZU Hiroaki
金额：
$ 1.98万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671157/
关键词：
major histocompatibility complex antigen gene transfection immunotherapy electroporation graft rejection

项目摘要

1. In our first study, we investigated immunogenicity of low-antigenic rat adenocarcinoma cells after transfection with an allogeneic class I major histocompatibility complex (MHC) gene in vitro, and studied whether direct allogeneic MHC gene transfer into the established tumor using in vivo electroporation might stimulate the host immune response, and also provide an immunotherapeutic effect. Mammary adenocarcinoma cells (MAT B III) originated from F344 rat (RT1AィイD11ィエD1) were transfected with a plasmid DNA encoding RT1AィイD1aィエD1 (pcMRT1A) in vitro. The expression of RT1AィイD1aィエD1 antigen on the tumor cells resulted in stimulating cytolytic T-cell response against specific gene products. Furthermore, we investigated the antitumor effects of direct allogeneic MHC class I gene transfer into the tumor grown in the syngeneic host using in vivo electroporation. Intratumoral injection of the pcMRT1A-Lipofectin complex followed by eight electrical pulses (99μsec, 400V/cm) through two electr … More odes located on each side of the tumor induced a marked regression in tumor growth and prolonged survival periods of the host. These results indicate that transferring allogeneic class I MHC gene directly into tumor using in vivo electroporation could induce a cell-mediated immune response and provide an immunotherapeutic effect for the established tumor.2. In our second study, we examined whether insertion of an antisense vector into the allograft could down-regulate the expression of the target antigens and to prevent the rejection of transplanted organs. In vitro study, down-regulation of donor MHC class I antigen expression on the donor cells was observed after transfection of plasmid vector containing antisense of donor target antigen in vitro and also immunological response to these cells. However, in vivo, efficient transfection of antisense DNA to the donor graft organ was very difficult at present. Now we are developing new technology to provide for efficient in vivo transfection. Less

1.在我们的第一项研究中，我们调查了低抗原性大鼠腺癌细胞与异基因I类主要组织相容性复合体（MHC）基因转染后的免疫原性，并研究了是否直接异基因MHC基因转移到建立的肿瘤使用体内电穿孔可能会刺激宿主免疫应答，也提供了免疫抑制作用。用编码RT 1A腺病毒D1 a腺病毒D1（pcMRT 1A）的质粒DNA体外转染F344大鼠（RT 1A腺病毒D11腺病毒D1）的乳腺癌细胞（MAT B III）。肿瘤细胞上RT 1A β D1 a β D1抗原的表达导致刺激针对特定基因产物的细胞溶解性T细胞应答。此外，我们研究了直接同种异体MHC I类基因转移到肿瘤生长在同基因宿主体内电穿孔的抗肿瘤作用。瘤内注射pcMRT 1A-Lipofectin复合物，然后通过两个电脉冲（99μsec，400 V/cm）进行8次电脉冲， ...更多信息位于肿瘤两侧的ODES诱导肿瘤生长显著消退并延长宿主的存活期。这些结果表明，利用体内电穿孔技术将同种I类MHC基因直接导入肿瘤组织，可以诱导细胞免疫应答，对已建立的肿瘤组织产生免疫抑制作用.在我们的第二项研究中，我们检查了将反义载体插入同种异体移植物中是否可以下调靶抗原的表达并防止移植器官的排斥反应。体外实验中，供体靶抗原反义核酸质粒载体转染供体细胞后，供体细胞MHC Ⅰ类抗原表达下调，并观察到供体细胞产生的免疫应答。然而，目前在体内，将反义DNA有效地转染到供体移植器官中是非常困难的。现在，我们正在开发新的技术，以提供有效的体内转染。少