EXPERIMENTAL ANALYSIS OF VASCULER CHANGES IN CHRONIC ALLOGRAFT REJECTION USING HUMANIZED SCID MOUSE MODEL.

慢性移植物肾病(CAN)是临床肾移植中最常见的移植失败原因。然而，由于CAN的病因和发病机制各不相同，目前对CAN的治疗策略尚不明确。CAN的基本病理生理学与导致纤维化和螺旋硬化的血管炎症有关。我们的研究目的是利用Hu-PBL-SCOD小鼠对CaN的动物模型进行分析和鉴定。以人外周血淋巴细胞(PBL)为供体，建立了具有人类免疫环境的人源化严重联合免疫缺陷(SCID)小鼠。这些HuPBL-SCID小鼠也被证明在它们的循环中涉及功能性的人类淋巴细胞和免疫球蛋白。在HU-PBL-SCID中，采用人肠系膜动脉直接转流腹主动脉，在小鼠体内展示了人类同种异体移植的真实情况。对修复的血管移植物的分析显示，淋巴细胞侵袭了血管移植物的内皮。病理分析表明，这些移植物浸润性淋巴细胞(GIL)在染色上属于CD3CD4。接下来，我们试图通过改变移植物的恢复时间来确定观察细胞排斥反应的最佳时间。然而，由于个体间的差异，很难建立稳定的细胞排斥模型。我们还试图通过将移植物回收到组织培养来建立体外GIL细胞系。然而，由于污染和GIL生长缓慢，这项实验导致了停产。应用适当的反义寡核苷酸的聚合酶链式反应分析发现，在恢复的人肠系膜动脉中有肿瘤坏死因子-α和干扰素-γ的信使RNA表达。另一方面，在我们的实验条件下，没有与细胞迁移或趋化因子相关的黏附分子的特定信使表达。

Chronic allograft nephropathy (CAN) is the most common cause to graft failure in clinical kidney transplantation. The appropriate therapeutic strategies to CAN, however, is yet to be established because of the various etiology and rationale of CAN. Basic pathophysiology of CAN is accounted for vascular inflammation leading to fibrosis and screlosis. Our study goal was to analyze and characterize CAN in animal model using hu-PBL-SCOD mouse. Humanized severe combined immunodeficient (SCID) mice, which exhibited human immunological environment, were established by administration of human peripheral blood lymphocytes (PBL). These huPBL-SCID mice were also demonstrated to involve functional human lymphocyte and immunoglobulin in their circulation. To display the real CAN of human allotransplant circumstances in mouse, human mesenteric arterior was applied for direct bypass of abdominal aorta in hu-PBL-SCID. Analysis of recovered vascular graft revealed lymphocytes invasion on the endothelium of the vascular graft. Pathological analysis indicated these graft-infiltrating-lymphocytes (GIL) belonged to CD3+CD4+ in staining. Next, we tried to determine the optimal time to see the cellular rejection by changing the recovering time of graft. Stable model of the cellular rejection were, however, difficult to establish due to the inter-individual differences. We also tried to establish the GIL cell line in vitro by recovering these grafts to tissue culture. This experiment, however, resulted in discontinued because of the contaminations and slow growing GILs. Messenger RNA expression of TNF-alpha and IFN-gamma were noted in recovered human mesenteric arterior using PCR analysis applying the appropriate anti-sense oligonucleotides. On the other hand, no particular messenger expression of adhesion molecules associated with cell migration or chemokines in our experimental condition.