Analysis of mechanisms in delayed xenograft and cellular rejection in xenotransplant between human and swine

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。这只HU-PBL-SCID小鼠也被证明在其循环中涉及功能性人类淋巴细胞和免疫球蛋白。在HU-PBL-SCID中，采用猪肠系膜动脉直接转流腹主动脉，以显示小鼠从猪到人的真实异种移植情况。对回收的异种移植物的分析显示，人IgM黏附于异种移植物的内缘，淋巴细胞侵袭异种移植物的内皮。通过免疫吸收阻断异种反应性的血清学反应，病理分析表明这些淋巴细胞在染色上属于人CD3+CD4+。接下来，我们试图通过改变异种移植的恢复时间来确定在最严重的情况下观察细胞排斥反应的最佳时间。稳定的细胞排斥反应模型。然而，难以进行ES…由于个体间的差异，更具表现力。在猪动脉异种移植物的侵袭过程中，未检测到人淋巴细胞。我们还尝试用辐照的猪主动脉内皮细胞(PAEC)作为刺激物，以辐照的原始人淋巴细胞作为饲养层，将该异种移植物恢复到组织培养中，以建立体外的人异种反应性细胞系。然而，由于移植物的污染，这个实验导致了不可能的情况。最近，我们的同事Sasaki博士发现，人类NK细胞识别异种移植中不表达的人类白细胞抗原-G和E基因，并在异种移植排斥反应中发挥了作用(移植67；1999年31-37)。我们试图在HU-PBL-SCID小鼠模型的活体系统中表明这一证据。HU-PBL-SCID小鼠移植的异种动脉未见抑制反应，但未见明显抑制作用。我们的一份包含这些结果的手稿现在(在印刷中)。较少

Humanized severe combined immunodeficient (SCID) mice, which exhibited human immunological environment, were established by administration of human peripheral blood lymphocytes (PBL). This hu-PBL-SCID mice were also demonstrated to involve functional human lymphocyte and immunoglobulin in their circulation. To display the real xenotransplant circumstances from swine graft to human in mouse, swine mesenteric arterior was applied for direct bypass of abdominal aorta in hu-PBL-SCID. Analysis of recovered xenograft revealed human IgM adherence to inner rim and lymphocytes invasion on the endothelium of the xenograft. By blocking the xenoreactive serological response with immuno-absorption, pathological analysis indicated these lymphocytes belonged to human CD3+CD4+ in staining. Next, we tried to determine the optimal time to see the cellular rejection in the severest condition by changing the recovering time of xenograft. Stable model of the cellular rejection were.however, difficult to es … More tablish due to the interindividual differences. Human lymphocytes were unable to be detected in the invasion into swine arterior xenografts. We also tried to contitute the human xenoreactive cell line in vitro by recovering this xenografts to tissue culture using irradiated pig aortic endothlial cell (PAEC) as stimulator and irradiated original human lymphocytes as feeder. This experiment, however, resulted in impossible condition due to contaminations from grafts.Recently Dr.Sasaki, one of our colleagues, showed that human NK cells recognized non-expression of HLA-G ＆ E gene in xenograft and played a role in xenograft rejection (Transplantation 67; 31-37 1999). We tried to indicate this evidence in in vivo system using hu-PBL-SCID mice model. Swine arterior xenogarfts transfected with HLA-G ＆ E gene were recovered from hu-PBL-SCID mice, but no suppressive effect of xenoreactive responses were noted in this experoment. One of our manuscripts containing these results were now (in press). Less