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: 6767770
• 负责人: Anthony P Monaco
• 金额: $ 45.02万
• 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/6767770
• 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）：慢性免疫抑制是相关的 感染，肿瘤，药物毒性，代谢作用和未能 控制拒绝。一种无毒的，容易应用的方法诱导公差是 临床移植的目标。我们产生了稳定的耐受性， 使用抗淋巴细胞血清（ALS），雷帕霉素和供体骨骼的宏观主义 完全主要的组织兼容复合物（MHC）中的骨髓（BM）不匹配 啮齿动物。 ALS和雷帕霉素是一种极有效的免疫抑制 组合; ALS诱导组织淋巴细胞耗竭和T细胞凋亡 雷帕霉素促进激活诱导的T细胞凋亡。我们寻求 证明该方案在cynomolgus猴子中的有效性 临床应用。在AIM 1中，接收者猴子将获得胸腺球质蛋白 （第-1至10天），雷帕霉素（第0至90天），肾脏同种异体移植（第0天），有OR 第10天没有捐助者的特定BM。接收者将遵循标准 肾功能测试和协议活检，并串行监测抗多onor 同种异体。组织淋巴细胞耗竭将由 组织学，FACS分析，T细胞凋亡和免疫反应性。外围 血液将被分析用于嵌合。抗抑制免疫将通过 反式体内DTH分析。耐受性的诱导和稳定性将相关 随着免疫反应性，嵌合的程度和持续时间的改变，以及 淋巴细胞耗竭的程度。另外，较高剂量的BM和/或 胸腺球蛋白和细胞因子（G-CSF）刺激外周血Budy涂层 细胞将在耐受诱导方面的有效性测试。 AIM 2将使用 RT -PCR和免疫组织化学以鉴定抗凋亡基因A20，HO-L ECK并将其存在与宽容相关联。 AIM 3将利用真实 时间RT -PCR识别各种免疫相关基因的表达为标记 用于亚临床排斥和基因芯片分析以识别基因模式 与公差相关的表达。这个基于非放射的协议 组合使用两种广泛使用的胸腺球蛋白和雷帕霉素 使用BM，作为一种易于应用，潜在耐受性良好的协议 临床耐受性诱导。