Tolerance In Organ Transplantation And Autoimmune Disease By T Cell Immunotoxin

T 细胞免疫毒素对器官移植和自身免疫性疾病的耐受性

• 批准号: 7594527
• 负责人: David M. Neville
• 金额: $ 113.17万
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 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/7594527
• 关键词: Adverse effects; Allografting; Antibodies; Antigen Presentation; Appearance; Autoimmune Diseases; B-Lymphocytes; Back; Binding; Binding Sites; Blood; Blood Glucose; CD3 Antigens; Cell Transplants; Chronic; Clinical; Clinical Research; Clinical Treatment; Cyclosporine; Cyclosporins; Diabetes Mellitus; Diphtheria Toxin; Epitopes; Exhibits; Glycosylated Hemoglobin; Graft Survival; Gusperimus; HIV; Hour; Human; Immunization; Immunosuppression; Immunotoxins; Institution; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Islets of Langerhans Transplantation; Kidney Transplantation; Knowledge; Macaca mulatta; Malignant Neoplasms; Mediating; Methylprednisolone; Modeling; Monkeys; Monoclonal Antibody HuM291; Muromonab-CD3; Numbers; Organ; Organ Transplantation; Phase I Clinical Trials; Pichia; Process; Production; Proteins; Protocols documentation; Reagent; Recombinant Antibody; Recombinants; Reporting; Resistance; Side; Solid Neoplasm; Specificity; Streptozocin; Structure; Survivors; T-Cell Depletion; T-Cell Leukemia; T-Lymphocyte; Therapeutic immunosuppression; Toxin; Transplantation; Viral; Yeasts; cell killing; cell type; chemical conjugate; day; desire; graft versus host disease induction; in vivo; islet; killings; lymph nodes; mutant; preclinical study; response

The general aim of this project is to study the effects of immunotoxin induced transient T-cell depletion on the modulation of T cells from immunization responses towards tolerizing responses, and to use this knowledge for the experimental and clinical treatment of T cell driven autoimmune diseases, graft-versus-host disease, and the induction of tolerance to mismatched organ and cell transplants. We previously showed that a 2-3 day course of anti-rhesus CD3 immunotoxin constructed with CRM9, a binding site mutant of diphtheria toxin, depletes lymph node and blood T cells by 99% after a 48 hour lag period. This process was associated with a marked prolongation of survival of mismatched functioning rhesus kidney transplants, 50% over 9 months without further immunosuppressive therapy. Rejections, were preceded by the appearance of anti-graft antibodies (done with S. Knechtle and J. Thomas). To circumvent this adverse T cell mediated B cell response that is apparently initiated during the delay in immunotoxin induced T cell killing, we have added a short course of agents that are believed to block antigen presentation such as deoxyspergualin (DSG). Out of 5 bilaterally rephrectomized recipients long term tolerance was achieved in 3 without further immunosuppresion as judged by a graft survival of 4 years or longer. Congeneic pancreatic islet cell transplants in monkeys suffering from spontaneous insulin dependent diabetes have been shown to reverse diabetes as judged by returning non-fasting blood glucose and glycosylated hemoglobin values to normal in the absence of exogenous insulin. A short course of immunotoxin, methyl prednisolone and cyclosporine provide induction of stable operational tolerance to islets that has lasted over one year in one case. This is the first reported reversal of diabetes using a tolerizing protocol that does not depend on chronic immunosuppression, a process previously shown to compromise transplanted islet function (done with J. Thomas). Streptozotocin induced diabetes in monkeys has been successfully reversed by using anti-T cell immunotoxin in combination with 14 days of DSG and allografted pancreatic islets (done with J. Thomas). Three out of seven are long term tolerant survivors (6-7 years). A recombinant divalent anti-human anti-T cell immunotoxin has been developed that has 20-fold the potency of the previous chemical conjugates. This is expressed in Pichia pastoris. This material has now been produced for a phase I clinical trial in T cell leukemia and an INDA has been filed. In order to aid the production of immunotoxin and other bioactive proteins in Pichia pastoris, new strains have been constructed that exhibit toxin resistance and increased heterologous protein production in the presence of an active unfolded-protein response. Recently, a recombinant anti-CD3 immunotoxin active against monkey T cells has been developed. This was done in order to facilitate pre-clinical studies in organ transplantation and autoimmune diseases. A side benefit of this study was that it required explorations of different recombinant antibody geometries for optimal activity. A fold-back diabody structure was found to be most optimal toward monkey CD3. Model building studies predicted that this structure should have very high binding activity towards targets that contain homodimeric epitopes rotated by 180 degrees. This prediction was found to be true in the case of a human solid tumor epitope and clinical studies at another institution are planned.

该项目的总体目标是研究免疫毒素诱导的短暂 T 细胞耗竭对 T 细胞从免疫反应到耐受反应的调节的影响，并将这些知识用于 T 细胞驱动的自身免疫性疾病、移植物抗宿主病以及诱导对不匹配的器官和细胞移植的耐受性的实验和临床治疗。我们之前表明，用 CRM9（白喉毒素的结合位点突变体）构建的抗恒河猴 CD3 免疫毒素的 2-3 天疗程，在 48 小时滞后期后可将淋巴结和血液 T 细胞消耗 99%。这一过程与功能不匹配的恒河猴肾移植的存活率显着延长有关，在不进行进一步免疫抑制治疗的情况下，9个月内存活率提高了50%。在拒绝之前，出现了抗移植物抗体（由 S. Knechtle 和 J. Thomas 完成）。为了避免这种不利的 T 细胞介导的 B 细胞反应（这种反应显然是在免疫毒素诱导的 T 细胞杀伤延迟期间启动的），我们添加了短期的药物，据信可以阻断抗原呈递，例如脱氧精胍菌素 (DSG)。根据移植物存活 4 年或更长时间来判断，5 名双侧肾切除受者中有 3 名在没有进一步免疫抑制的情况下实现了长期耐受。患有自发性胰岛素依赖型糖尿病的猴子的同种胰岛细胞移植已被证明可以逆转糖尿病，这是通过在不存在外源胰岛素的情况下将非空腹血糖和糖化血红蛋白值恢复到正常来判断的。短期疗程的免疫毒素、甲基泼尼松龙和环孢菌素可诱导对胰岛的稳定操作耐受，在一个病例中这种耐受持续了一年多。这是首次报道使用不依赖于慢性免疫抑制的耐受方案逆转糖尿病，此前已证明慢性免疫抑制会损害移植的胰岛功能（由 J. Thomas 完成）。通过使用抗 T 细胞免疫毒素并结合 14 天的 DSG 和同种异体移植胰岛（由 J. Thomas 完成），链脲佐菌素诱导的猴子糖尿病已成功逆转。 七分之三是长期耐受的幸存者（6-7 年）。一种重组二价抗人抗 T 细胞免疫毒素已被开发出来，其效力是之前化学缀合物的 20 倍。这在毕赤酵母中得到表达。该材料现已生产用于 T 细胞白血病的 I 期临床试验，并已提交 INDA。为了帮助巴斯德毕赤酵母产生免疫毒素和其他生物活性蛋白，我们构建了新菌株，在存在活性未折叠蛋白反应的情况下表现出毒素抗性并增加异源蛋白产量。 最近，开发出了一种对猴 T 细胞具有活性的重组抗 CD3 免疫毒素。 这样做是为了促进器官移植和自身免疫性疾病的临床前研究。 这项研究的一个附带好处是，它需要探索不同的重组抗体几何形状以获得最佳活性。 发现折回双抗体结构对于猴 CD3 是最优化的。 模型构建研究预测，该结构对包含旋转 180 度的同二聚体表位的靶标应具有非常高的结合活性。 这一预测在人类实体瘤表位的情况下被发现是正确的，并且计划在另一个机构进行临床研究。

项目成果

T-cell depletion and graft survival induced by anti-human CD3 immunotoxins in human CD3epsilon transgenic mice.

人 CD3ε 转基因小鼠中抗人 CD3 免疫毒素诱导 T 细胞耗竭和移植物存活。

• DOI: 10.1097/00007890-200205270-00023
• 发表时间: 2002
• 期刊: Transplantation
• 影响因子: 6.2
• 作者: Weetall,Marla;Digan,MaryEllen;Hugo,Ronald;Mathew,Sheba;Hopf,Christine;Tart-Risher,Nicole;Zhang,Jin;Shi,Victor;Fu,Fumin;Hammond-McKibben,Denise;West,Susan;Brack,Richard;Brinkmann,Volker;Bergman,Reinhard;NevilleJr,David;Lake,
• 通讯作者: Lake,

Chimeric fusion proteins--diphtheria toxin-based.

嵌合融合蛋白——基于白喉毒素。

• 发表时间: 2001
• 期刊: Current opinion in investigational drugs (London, England : 2000)
• 作者: Frankel,AE;Powell,BL;Vallera,DA;NevilleJr,DM
• 通讯作者: NevilleJr,DM

Effects of xenogeneic thymic transplantation in baboons.

异种胸腺移植对狒狒的影响。

• DOI: 10.1016/s0041-1345(00)02244-2
• 发表时间: 2001
• 期刊: Transplantation proceedings
• 影响因子: 0.9
• 作者: Wu,A;Yamada,K;Awwad,M;Kitamura,H;Wain,J;Neville,DM;Cooper,DK;Sykes,M;Sachs,DH
• 通讯作者: Sachs,DH

Xenogeneic thymus transplantation in a pig-to-baboon model.

猪狒狒模型中的异种胸腺移植。

• DOI: 10.1097/01.tp.0000044137.97841.99
• 发表时间: 2003
• 期刊: Transplantation
• 影响因子: 6.2
• 作者: Wu,Anette;Yamada,Kazuhiko;Neville,DavidM;Awwad,Michel;Wain,JohnC;Shimizu,Akira;Gojo,Satoshi;Kitamura,Hiroshi;Colvin,RobertB;Cooper,DavidKC;Sykes,Megan;Sachs,DavidH
• 通讯作者: Sachs,DavidH

Minimization of aggregation of secreted bivalent anti-human T cell immunotoxin in Pichia pastoris bioreactor culture by optimizing culture conditions for protein secretion.

通过优化蛋白质分泌的培养条件，最大限度地减少毕赤酵母生物反应器培养中分泌的二价抗人 T 细胞免疫毒素的聚集。

• DOI: 10.1016/j.jbiotec.2005.07.004
• 发表时间: 2006
• 期刊: Journal of biotechnology
• 影响因子: 4.1
• 作者: Woo,JungHee;Liu,YuanYi;NevilleJr,DavidM
• 通讯作者: NevilleJr,DavidM