Research about mechanism of delayd xenograft rejection (DXR).

延迟异种移植排斥反应（DXR）机制的研究。

• 批准号: 07457261
• 负责人: OKA Takahiro
• 金额: $ 4.16万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457261/
• 关键词: xenotransplantation; delayd xenograft rejection; NK cell; perforin; granzyme; CVF (Cobra Venom Factor); 遅延型異移拒絶反応; CVF (Cobra Venom Factor); discordant異種移植; sCR1; AT-III; CVF

Purpose : Delayd xenograft rejection (DXR) takes place in discordant xenotransplantation, even complement activation, natural antibody and blood coagulation are inhibited by drugs such as CVF (cobra venom factor). Although pathology of the heart underlying DXR shows infiltration of NK (natural killer) cells and macrophages, the mechanism of how these cells can induce DXR is unknown. In this study, we addressed the contribution of NK cells to DXR using congenitally NK activity deficient rat, Beige rat (Bg) that is derived from DA rat (DA).Materials and methods :1. NK activity (^<51>Cr release assay) was measured using Yac-1 cells as targets and splenocytes of DA and Bg as effector cells.2. Expression of cell surface antigens in DA and Bg splenocytes was compared by flowcytemetry.3. Using RT-PCR,the expression of effector molecules such as perforin, granzyme A and granzyme B was detected in DA and Bg splenocytes.4. Under complement inhibition by CVF (-1,0 day. 30U/kg. iv.), guinea pig hearts were transplanted into DA and Bg, and their grafts survival time were compared.Result :1. NK activity in BG was severely impaired in comparison with that in DA.(DA : 62.8%, BG : 18.2% at E/T=100)2. There was no difference in NK cells number between DA and BG,whereas cell numbers of CD8 positive and IL2R positive cells were decreased in Bg as compared with those in DA.3. The expressions of perforin, granzyme A and granzyme B mRNA were inhibited in Bg as compared with those in DA.4. Grafts mean survival time in Bg (28(]SY.+-。[)1.5hr, n=9) was significantly longer than that in DA (20(]SY.+-。[)2.1hr, n=9).Conclusion : NK cells and CD8 positive cells play an important role in the process of DXR.Therefore, the inhibition of these cells function would lead to longer xenografts survival.

目的：延迟异种移植排斥（DXR）发生在不协调的异种移植中，即使补体激活、天然抗体和凝血也会被 CVF（眼镜蛇毒因子）等药物抑制，尽管 DXR 的心脏病理显示 NK（自然杀伤细胞）浸润。 ）细胞和巨噬细胞，这些细胞如何诱导 DXR 的机制尚不清楚。在这项研究中，我们利用 NK 细胞对 DXR 的贡献进行了研究。先天性NK活性缺陷大鼠，源自DA大鼠(DA)的米色大鼠(Bg)。 材料和方法：1．使用Yac-1细胞作为靶标和脾细胞测定NK活性(^ 51 Cr释放测定)。 DA和Bg作为效应细胞。2．流式细胞术比较DA和Bg脾细胞表面抗原的表达。3． DA和Bg脾细胞中检测到穿孔素、颗粒酶A和颗粒酶B。4．CVF(-1.0天，30U/kg，静脉注射)补体抑制下，将豚鼠心脏移植到DA和Bg及其移植物中。比较生存时间。结果：1.与DA相比，BG中的NK活性严重受损。（DA：62.8％，BG： E/T=100时为18.2%)2．DA和BG中NK细胞数量无差异，而Bg中CD8阳性和IL2R阳性细胞数量较DA有所减少。3．与 DA 中的相比，Bg 中的穿孔素、颗粒酶 A 和颗粒酶 B mRNA 受到抑制。4. Bg 中的移植物平均存活时间 (28(]SY.+-. [)1.5hr, n=9)明显长于DA (20(]SY.+-.[)2.1hr, n=9)。结论：NK细胞和CD8阳性细胞在此过程中发挥重要作用因此，抑制这些细胞功能将导致异种移植物存活时间更长。

项目成果

I.Fujiwara,H.Nakajima: "Prolongation of Discordant Xenograft Survival by sCRI and AT-III Combination Therapy" Transplantation Proceedings. (in press).

I.Fujiwara、H.Nakajima：“通过 sCRI 和 AT-III 联合疗法延长不一致的异种移植物存活”移植论文集。

Hiroo Nakajima, Takahiro Oka: "The Inhibition of TcR-Induced Fas-ligand Upregulation by CsA and FK506" Transplant Proc.Vol. 28, No. 2. 1052-1055 (1996)

Hiroo Nakajima、Takahiro Oka：“CsA 和 FK506 对 TcR 诱导的 Fas 配体上调的抑制”移植 Proc.Vol。

Hiroo Nakajima, et al: "The Inhibition of TcR-Induced FasL Upregulation by CsA and FK506" Transplantation Proceedings. Vol28/No2. 1052-1055 (1996)

Hiroo Nakajima 等人：“CsA 和 FK506 对 TcR 诱导的 FasL 上调的抑制”移植论文集。

岡隆宏,藤原郁也: "異種移植と遺伝子制御" 今日の移植. Vol,8. 321-326 (1995)

Takahiro Oka，Ikuya Fujiwara：“异种移植和基因控制”《今日移植》卷，8. 321-326 (1995)。

Ikuya Fujiwara, Hiroo Nakajima, et al: "Prolongation of Discordant Xenograft Survival by sCR1 and AT-3 Combination Therapy" (in press).

Ikuya Fujiwara、Hiroo Nakajima 等人：“通过 sCR1 和 AT-3 联合疗法延长不一致异种移植物的存活率”（待出版）。