权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

A study for the in vivo mechanism of transplantation tolerance using GFP transgenic mice

GFP转基因小鼠体内移植耐受机制研究

基本信息

批准号：
10470274
负责人：
SHIRAKURA Ryota
金额：
$ 4.61万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
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-10470274/
关键词：
Transplantation tolerance T-cells Mice

项目摘要

The Green Fluorescent Protein (GFP) transgenic mice with B6 background, bm1, or bm12 mice were cleaned up by egg-transfer technique and specific pathogen-free mice were generated. First, whether the GFP acts as transplantation antigen was tested by skin-grafting and we found it only worked as very weak transplantation antigens (i.e.: mean survival>60 days). Thus, GFP transgene are used as marker to track allogeneic cells in vivo in the following studies. Second, the following findings were made by the skin-grafting using non transgenic mice.A : The survival of class I MHC-disparate bm1 skin grafts were dose-dependently prolonged by intravenous presensitization of B6 host with one to ten million of(B6×bm1)F1 spleen cells.B : Intravenous presensitization of B6 host with even a hundred million of (B6×bm12) F1 spleen cells did not have any significant effect on the survival of class II MHC-disparate bm12 skin grafts.Third, the fate of class I or class II MHC-disparate allogeneic cells inje … More cted into B6 host was tracked in vivo using GFP-positive B6, (B6×bm1) F1, or (B6×bm12) F1 spleen cells. Ten million spleen cells from each mice were intravenously injected and the host spleen cells were analyzed with FACS 1, 2, 4, or 8 weeks after the injection. The following results were achieved by the investigation.C : The injected B6 cells consisted of 1-2% of host spleen cells at 1 weeks and the fraction of the injected cells detected in hosts were proportional to those before the injection. The number of injected cells detected gradually decreased to 0.2-0.3% by 4 weeks and became undetectable 8 weeks after the injection. The speed of decay seemed not to be affected by the type of cells injected.D : The class I MHC-disparate (B6×bm1) F1 cells injected were detected at 1 and 2 weeks after the injection and became undetectable at 4 weeks and thereafter. The numbers of cells detected were constantly smaller than those of injected B6 cells but the speed of decay seemed unaffected by the type of cells detected.E : The total number of class II MHC-disparate (B6×bm12) F1 cells injected was roughly similar to those of (B6×bm1) F1 cells at 1, 2, 4, or 8 weeks after the injection. However, class II MHC-disparate B cells were disappeared more quickly than the other types of cells included in spleen.In summary, although we could not perform all the studies planned because of the delay in cleaning-up process of the mice, we could get some interesting results from the novel method which has the potential to clarify the undiscovered mechanisms of transplantation tolerance. In addition, the other molecular biological methods established in this study such as quantitative RT-PCR proved to be useful in the other fields of investigation and the three papers using those methods were published as listed on the other side of the form. Less

用卵子转移技术对B6背景的绿色荧光蛋白(GFP)转基因小鼠、bm1、bm12转基因小鼠进行清理，获得无特定病原体的小鼠。首先，通过皮肤移植检测GFP是否作为移植抗原，我们发现它只作为非常弱的移植抗原发挥作用(即：平均存活60天)。因此，在接下来的研究中，GFP转基因被用作体内示踪同种异体细胞的标记。B：1~1000万个(B6×bm1)F1脾细胞静脉预敏化B6宿主对移植物的存活无显著影响。3、…中I类或II类异基因bm12皮片的存活用GFP阳性的B6、(B6×bm1)F1或(B6×bm12)F1脾细胞在体内追踪更多进入B6宿主的细胞。每只小鼠静脉注射1000万个脾细胞，在注射后1、2、4或8周用流式细胞仪对宿主脾细胞进行分析。研究结果如下：1周时，注射的B6细胞占宿主脾细胞的1-2%，在宿主体内检测到的注射细胞比例与注射前成正比。注射后4周检测到的细胞数逐渐减少到0.2-0.3%，8周后检测不到。细胞的衰退速度似乎不受注入的细胞类型的影响。D：注入的I类MHC不同的(B6×bm1)F1细胞在注射后1周和2周检测到，4周后检测不到。检测到的细胞数量始终少于注射的B6细胞，但衰退的速度似乎不受检测到的细胞类型的影响。E：注射II类MHC不同的(B6×bm12)F1细胞的总数在注射后1、2、4或8周与(B6×bm1)F1细胞大致相同。综上所述，尽管由于小鼠清理过程的延迟，我们不能进行所有计划中的研究，但我们可以从新方法中获得一些有趣的结果，它有可能阐明移植耐受的未知机制。此外，本研究中建立的其他分子生物学方法，如定量RT-PCR，在其他研究领域也被证明是有用的，使用这些方法的三篇论文发表在表格的另一面。较少