Induction of transplantation tolerance by co-transplantation of vascularized thymic lobe in miniature swine

小型猪血管化胸腺叶联合移植诱导移植耐受

• 批准号: 15390387
• 负责人: SADA Masaharu
• 金额: $ 9.15万
• 依托单位: National Cardiovascular Center, Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390387/
• 关键词: SLA; miniature swine; tolerance; vascularized thymic lobe; CAV; MHC; SLA class I, II遺伝子; PCR-SSP; PCR-RFL; miniature pig; SLA class I, II; PCR-RFLP; genotyping; RT-PCR; SBT

To induction of tolerance for organ transplantation, we have performed on analysis of SLA antigens in miniature swine, full SLA antigens mismatched organ transplantation and co-transplantation of vascularized thymic lobe administrated short-term immunosuppression in miniature swine. In inbred Clawn miniature swine, three haplotypes have been assined by RT-PCR of SLA class I and II genes. To determine of SLA antigens of Clawn miniature swine, we have developed a polymerase chain reaction-sequence specific primer (PCR-SSP) method for SLA class I (SLA-1,-2,-3 genes) and SLA class II (DRB1 gene), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for SLA class II (DQB1 gene). Based on allele analysis of SLA class I and II genes, eleven different phenotype pattern and two new haplotypes were identified of this study. We have established pure-bred Clawn miniature swine, twenty-two of identified SLA gene homozygotes and nineteen of heterozygotes. We have … More transplanted seventeen hearts heterotopically across either minor mismatched or MHC-class II mismatched MHC-inbred miniature swine to assess (1)whether a short course of FK560 induces tolerance and prevents the development of cardiac allograft vasculopathy (CAV), and (2)whether a new immunosuppressant, FK778,which has been shown to have inhibitory effects on the cellular and humoral immune responses. The 12-day course of FK506 failed to induce tolerance in this model, but it did prolong the mean graft survival across a MHC-class II mismatched barrier compared with control model. The co-administration of FK778 with FK506 further prolong the mean graft survival, induced donor specific hyporesponsineness in vitro, and inhibited the development of CAV. We have previously reported the ability of vascularized thymic lobe (VTL) allografts to induce transplantation tolerance across full MHC mismatch in thymectomized miniature swine. In this study, we have investigated the mechanism of long-term acceptance in recipient swine following cotransplantation of VLT in fully MHC-mismatched cardiac transplantation heterotopically. All recipients that received a VTL maintained their grafts significantly longer than only heart graft, and those receiving 28days of tacrolimus maintained their heart graft long-term. Cytotoxic T-lymphocyte responses against third-party antigens by cells from tolerant animals showed restriction by both self and donor MHC, whereas responses of controls were restricted to self MHC only. The presence of donor dendritic cells in VTL grafts and results of co-culture assays suggested both central and regulatory mechanisms were involved an achieving long-term acceptance. Less

为诱导器官移植耐受，我们进行了小型猪SLA抗原分析、全SLA抗原错配器官移植和带血管胸腺叶短期免疫抑制联合移植。用SLA-I和SLA-II类基因的RT-PCR方法对近交系克朗小型猪进行了三种单倍型的鉴定。建立了SLA I类基因(SLA-1、2、3基因)和SLA II类基因(DRB1基因)的聚合酶链式反应-序列特异性引物(PCR-SSP)方法和SLA II类基因(DQB1基因)的聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法。基于SLA I和II类基因的等位基因分析，本研究鉴定了11种不同的表型模式和2种新的单倍型。我们建立了纯种克劳恩小型猪，鉴定出SLA基因纯合子22头，杂合子19头。我们有…为了评估(1)短疗程FK560是否能诱导耐受并防止心脏移植物血管病(CAV)的发生，以及(2)一种新的免疫抑制剂FK778是否对细胞和体液免疫反应有抑制作用，我们对17只心脏移植患者进行了更多的异位移植，无论是轻微的不匹配还是MHC-II类不匹配的小型猪。FK506的12天疗程未能在该模型中诱导耐受，但与对照模型相比，它确实延长了跨越MHC-II类错配屏障的移植物平均存活率。FK778和FK506联合应用可进一步延长移植物的平均存活时间，在体外诱导供者特异性低反应，并抑制CAV的发展。我们之前已经报道了带血管胸腺叶(VTL)同种异体移植在胸腺切除的小型猪中诱导移植耐受的能力，跨越完全MHC不匹配。在这项研究中，我们研究了在完全MHC不相合的异位心脏移植中，VLT联合移植后受体猪长期接受的机制。所有接受VTL的受者的移植物维持时间明显长于仅接受心脏移植的受者，而接受28天他克莫司治疗的受者移植心脏的维持时间较长。耐受动物细胞对第三方抗原的细胞毒性T淋巴细胞反应受自身和供体MHC的限制，而对照组仅受自身MHC的限制。供者树突状细胞在VTL移植物中的存在和共培养试验的结果表明，中枢和调节机制都涉及实现长期接受。较少

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