Induction of allograft tolerance in non-human primates

非人类灵长类动物同种异体移植耐受的诱导

• 批准号: 6478662
• 负责人: Anthony P Monaco
• 金额: $ 44.19万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6478662
• 关键词: Macaca fascicularis; SCID mouse; T lymphocyte; apoptosis; artificial immunosuppression; bone marrow transplantation; cooperative study; flow cytometry; gene expression; genetic markers; histology; immune tolerance /unresponsiveness; immunocytochemistry; immunosuppressive antileukocyte serum; kidney transplantation; microarray technology; polymerase chain reaction; sirolimus

DESCRIPTION (provided by applicant): Chronic immunosuppression is associated with infection, neoplasms, drug toxicity, metabolic effects and failure to control rejection. A non-toxic, easily applied method to induce tolerance is a goal of clinical transplantation. We produced stable tolerance and macrochimerism using anti-lymphocyte serum (ALS), rapamycin and donor bone marrow (BM) in fully major histocompatability complex (MHC) mismatched rodents. ALS and rapamycin is an extremely effective immuno-suppressive combination; ALS induces tissue lymphocyte depletion and T cell apoptosis while rapamycin promotes activation-induced T cell apoptosis. We seek to demonstrate the effectiveness of this protocol in cynomolgus monkeys prior to clinical application. In Aim 1, recipient monkeys will receive Thymoglobulin (day -1 to 10), rapamycin (day 0 to 90), renal allografts (day 0), with or without donor specific BM on day 10. Recipients will be followed by standard renal function tests and protocol biopsies and monitored serially for antidonor alloreactivity. Tissue lymphocyte depletion will be determined by histology, FACS analysis, T cell apoptosis and immune reactivity. Peripheral blood will be assayed for chimerism. Anti-donor immunity will be assayed by trans vivo DTH assays. Induction and stability of tolerance will be correlated with alterations in immune reactivity, degree and duration of chimerism, and extent of lymphocyte depletion. In addition, higher doses of BM and/or thymoglobulin, and cytokine (G-CSF) stimulated peripheral blood buffy coat cells will be tested for effectiveness in tolerance induction. Aim 2 will use RT -PCR and immunohistochemistry to identify anti-apoptotic genes A20, HO-l and Eck and correlate their presence with tolerance. Aim 3 will utilize real time RT -PCR to identify expression of various immune related genes as markers for subclinical rejection and gene chip analysis to identify patterns of gene expression associated with tolerance. This non-radiation based protocol employs two widely used agents, thymoglobulin and rapamycin, in combination with BM, as an easily applied, potentially well tolerated protocol for clinical tolerance induction.

描述（由申请人提供）：与慢性免疫抑制相关 感染、肿瘤、药物毒性、代谢影响和失败 控制拒绝。一种无毒、易于应用的诱导耐受的方法是 临床移植的目标。我们生产了稳定的公差和 使用抗淋巴细胞血清 (ALS)、雷帕霉素和供体骨进行大嵌合 主要组织相容性复合体 (MHC) 中的骨髓 (BM) 完全不匹配 啮齿动物。 ALS 和雷帕霉素是一种极其有效的免疫抑制药物 组合; ALS 诱导组织淋巴细胞耗竭和 T 细胞凋亡 而雷帕霉素则促进激活诱导的T细胞凋亡。我们力求 证明该方案在食蟹猴中的有效性 临床应用。在目标 1 中，受体猴将接受胸腺球蛋白 （第-1天至第10天），雷帕霉素（第0天至第90天），肾同种异体移植物（第0天），用或 第 10 天没有捐赠者特定的 BM。接受者将遵循标准 肾功能测试和方案活检并连续监测抗捐赠者 同种异体反应性。组织淋巴细胞的消耗将通过以下方式确定 组织学、FACS 分析、T 细胞凋亡和免疫反应性。周边 将检测血液的嵌合状态。抗供体免疫力将通过以下方法测定 体内 DTH 测定。耐受性的诱导和稳定性将相关 免疫反应性、嵌合程度和持续时间的改变，以及 淋巴细胞耗竭的程度。此外，更高剂量的 BM 和/或 胸腺球蛋白和细胞因子 (G-CSF) 刺激外周血血沉棕黄层 将测试细胞在耐受诱导中的有效性。目标2将使用 RT-PCR和免疫组织化学鉴定抗凋亡基因A20、HO-1 和埃克并将它们的存在与耐受性联系起来。目标 3 将利用真实 时间RT-PCR来鉴定各种免疫相关基因的表达作为标记 用于亚临床排斥和基因芯片分析以识别基因模式 与耐受性相关的表达。这种非基于辐射的协议 联合使用两种广泛使用的药物：胸腺球蛋白和雷帕霉素 BM，作为一种易于应用、潜在耐受性良好的方案 临床耐受诱导。