Induction of cardiac allograft tolerance in a rat heart transplant model

在大鼠心脏移植模型中诱导心脏同种异体移植耐受

• 批准号: 9549480
• 负责人: Michael Solomon
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9549480
• 关键词: Acute; Adoptive Transfer; Alloantigen; Allograft Tolerance; Allografting; Animals; Bone Marrow Transplantation; Cell Therapy; Cells; Clinical; Cyclosporine; Data; Data Analyses; Dose; Genes; Genomics; Genotype; Goals; Heart; Heart Transplantation; Histocompatibility Antigens; Immune Tolerance; Immune system; Immunologic Markers; Immunologics; Immunosuppression; Immunosuppressive Agents; Inbred BN Rats; Incidence; Individual; Infection; Injectable; Injection of therapeutic agent; Laboratories; Laboratory Animals; Life; Literature; Lymphocyte; Lymphoid Cell; Maintenance Therapy; Malignant Neoplasms; Manuscripts; Methods; Modeling; Molecular Profiling; Morbidity - disease rate; Oligonucleotide Microarrays; Organ Donor; Organ Transplantation; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phenotype; Process; Proteins; Proteomics; Protocols documentation; Publishing; RNA; Rattus; Regulatory T-Lymphocyte; Reporting; Risk; Sampling; Serum; Sirolimus; Solid; Spleen; Splenocyte; T-Lymphocyte; Techniques; Testing; Th2 Cells; Therapeutic immunosuppression; Thymus Gland; Transplant Recipients; Transplantation; base; candidate marker; computerized data processing; cost; cytokine; density; design; heart allograft; immunological intervention; improved outcome; mortality; novel; standard of care

Although the immunosuppressive drugs currently in use are effective in reducing the incidence of acute rejection after transplantation, they also put the patient at risk for life threatening infections and cancers. The long term administration of immunosuppressive agents results in an increase in both morbidity and mortality. In addition, the long term cost of these agents represents a financial burden. Establishment of tolerance to a well-functioning transplant without nonspecific immunosuppressive drugs is a major goal of organ transplantation therapy. The induction of recipient (host) tolerance to the histocompatibility antigens of the organ donor could eliminate the need for long term administration of these nonspecific immunosuppressive drugs. This would have a major impact on the quality and quantity of life of patients with long term surviving organ grafts by reducing the immunologic and non-immunologic complications associated with long term immunosuppressive therapy. Induction of immunotolerance in animal heart transplant models has for the most part involved donor-based tolerance. One well established model is injection of donor splenocytes plus a single injection of the T-cell supressing agents anti-rat lymphocyte serum or RIB 5/2. Adoptive transfer (altering the hosts immune system using lymphoid cells from another individual) of ex vivo generated alloantigen-specific regulatory T cells induces immunotolerance in a rat transplant model. This method has also been used in bone marrow transplantation and can be the basis for recipient-based immunotolerance induction. In heart transplantation, recipient-based immunotolerance induction is clinically more applicable than donor-based since donor genotype is rarely known prior to transplant. To date there is limited data on recipient-based immunotolerance induction in heart transplantation, but data exists from the bone marrow transplant literature. In addition, although allograft tolerance has been achieved using a wide variety of donor-based immunologic interventions in laboratory animals, no reliable method of confirming tolerance following cardiac transplantation has been established. Standard of care in the field of solid organ transplantation remains initial treatment with triple immunosuppression followed by long term maintenance therapy with 1-3 immunosuppressive agents. One cannot withdraw drugs confidently even if the graft seems to be tolerant. This study will apply high throughput expression profiling with the goal of studying donor and recipient-based immunotolerance induction protocols and identifying protein and gene changes that could serve as possible candidate biomarkers of tolerance. Identifying laboratory methods that will permit safe and precise confirmation of immune tolerance in the transplant patient has the potential to improve outcome substantially. The protocol was approved in 2006 and a total of 403 rats were used over the duration of the protocol which was closed in 2012. The protocol was divided into 2 parts: Part 1 (recipient-based tolerance); and Part 2 (donor-based tolerance). In Part 1 (recipient-based tolerance), Stage 1 we were able to successfully generate Th2.rapa cells from recipient BN rats. In Part 1, Stage 2 these adoptively transferred ex vivo generated BN Th2 cells were tested in culture using flow and cytokine phenotype tests, and an optimal rapamycin dose was determined. In 2009, we completed studies in Part 1 Stage 3, which was designed to determine if Th2-shifted hosts (recipient-based tolerance) have reduced rejection. In Part 2 (donor-base tolerance), Stage 1 (Induction donor-based tolerance) we successfully learned the techniques to remove the spleen of the rat (DA) that will donate the heart in stage 2. We also successfully injected its processed splenocytes into the thymus of the rat (BN) that will receive the donor heart in stage 2. In 2009, we completed studies in Part 2, Stage 2 which were designed to determine if donor-based tolerance induction reduces rejection. In 2010, total RNA was prepared from peripheral blood mononuclear cells and used for high density oligonucleotide microarrays. A manuscript was published in 2011 (PloS One, 6(4): e18885, 2011) establishing the ability of host-type Th2. Rapa cell therapy given pre-transplant to shift post-transplant cytokines towards a Th2 phenotype and prolong allograft viability when used in combination with a short course of cyclosporine therapy. As mentioned above, in 2012 the protocol was closed; however, we continue to report on the project, since as new genomic techniques become available we may do further data processing and analysis of these completed studies and their stored samples.

尽管目前正在使用的免疫抑制药物可有效降低移植后急性排斥的发生率，但它们也使患者面临着威胁生命的感染和癌症的风险。免疫抑制剂的长期给药会导致发病率和死亡率增加。此外，这些代理商的长期成本代表了财务负担。 建立对无特异性免疫抑制药物的功能良好移植的耐受性是器官移植疗法的主要目标。对器官供体的组织相容性抗原的诱导（宿主）耐受性可以消除这些非特异性免疫抑制药物的长期施用。通过减少与长期免疫抑制治疗相关的免疫学和非免疫并发症，这将对具有长期存活器官移植的患者的寿命质量和数量产生重大影响。 动物心脏移植模型中免疫耐受性的诱导大部分涉及基于供体的公差。一个良好的模型是注射供体脾细胞，以及单次注射T细胞替代剂抗鼠淋巴细胞血清或肋骨5/2。过继转移（使用来自另一个个体的淋巴样细胞改变宿主的免疫系统）产生同种抗原特异性调节性T细胞可在大鼠移植模型中诱导免疫耐受性。该方法也已用于骨髓移植中，可以成为基于受体的免疫耐诱导的基础。在心脏移植中，基于受体的免疫耐诱导在临床上比基于供体的基因型更适用，因为供体基因型在移植前很少知道。迄今为止，心脏移植中基于受体的免疫耐诱导的数据有限，但数据来自骨髓移植文献。 此外，尽管使用了实验动物中各种基于供体的免疫干预措施来实现同种异体移植的耐受性，但尚未确定心脏移植后耐受性的可靠方法。固体器官移植领域的护理标准仍然是通过三重免疫抑制的初始治疗，然后使用1-3个免疫抑制剂进行长期维持治疗。即使移植物似乎是宽容的，也无法自信地提取药物。 这项研究将采用高吞吐量表达分析，以研究供体和基于受体的免疫耐诱导方案，并鉴定蛋白质和基因变化，这些蛋白质和基因变化可以作为耐受性的候选生物标志物。识别将允许安全，精确确认移植患者免疫耐受性的实验室方法，有可能大大改善结果。 该方案于2006年获得批准，并在2012年关闭的协议期间总共使用了403只大鼠。该协议分为2部分：第1部分（基于受体的公差）；和第2部分（基于捐助者的公差）。在第1部分（基于受体的公差）中，第1阶段我们能够成功地从受体BN大鼠中产生Th2.Rapa细胞。 在第1部分中，使用流量和细胞因子表型测试在培养物中测试了这些经过的离体产生的BN Th2细胞，并确定了最佳的雷帕霉素剂量。 在2009年，我们在第1阶段第3阶段完成了研究，该研究旨在确定Th2偏移的宿主（基于受体的公差）是否减少了排斥反应。 在第2部分（供体基本容忍度）中，第1阶（基于诱导供体的容忍度），我们成功地学习了去除将在第2阶段捐赠心脏的大鼠脾脏的技术。我们还成功地注入了其处理的脾脏的脾脏（BN），该阶段将在2009年中置于2阶段的阶段。耐受性诱导减少了排斥。 2010年，从外周血单核细胞中制备总RNA，并用于高密度寡核苷酸微阵列。 一份手稿于2011年出版（PLOS One，6（4）：E18885，2011），建立了宿主型TH2的能力。 Rapa细胞疗法将移植前移植后移植后细胞因子转移到Th2表型，并延长同种异体移植的生存能力，并与短期的环孢菌素治疗相结合。 如上所述，该协议在2012年关闭；但是，我们继续报告该项目，因为随着新的基因组技术的可用，我们可能会对这些完成的研究及其存储的样本进行进一步的数据处理和分析。