Immune reconstitution following autologous and allogeneic stem cell transplant

自体和同种异体干细胞移植后的免疫重建

• 批准号: 8158336
• 负责人: Frances Hakim
• 金额: $ 62.85万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 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/8158336
• 关键词: Allogenic; Autologous; Immune; Stem cell transplant; reconstitution

The Preclinical Service Core (PSC) projects have developed from transplantation protocols implemented by the clinical staff of ETIB. Using peripheral blood and marrow, and tumor and CGVHD tissue biopsies, we have evaluated lymphocyte subsets, cytokine content, T cell receptor repertoire diversity and thymopoietic activity. All data are incorporated into protocol-specific spreadsheets, linking samples to protocol arms and transplant time points, and are accessible by branch clinicians over secure NIH networks. Effects of IL-7 administration: In association with a Phase I clinical trial of IL-7 administration (P. I. Claude Sportes: 03-C-0152, in collaboration with Crystal Mackall in POB), we determined that IL-7 significantly expanded naive and central memory CD4 and CD8 T cells, as compared to effector cells, resulting in a significant increase in the diversity of the overall T cell receptor repertoire. These data were published in 2009 (Sportes et al 2009). In further studies of the clinical effects of this therapy, we determined that IL-7 produced a transient expansion of early B cell lymphopoiesis that was evident in the peripheral blood as an increase in transitional (T1) immature B cells. These data were incorporated into a report on the clinical effects of IL-7 therapy in 2010 (Sportes et al 2010). These studies support the use of IL-7 therapy to enhance T cell numbers and repertoire in patients with limited thymopoietic capacity, such as in aging populations and in patients with immune deficits after chemotherapy. Continuing studies will focus on the role of IL-7 in conjunction with vaccine responses in older patients. Immune reconstitution following intensive lymphodepletion and autologous CD34+ stem cell transplantation in patients with severe systemic lupus erythematosus (SLE): In an ongoing trial (P.I. Steven Pavletic 04-C-0095), T and B cell immune reconstitution has been monitored to assess the efficacy of the cytoreductive regimen in depleting T and B cells and the timecourse of immune recovery. These continuing studies contribute to development of autologous transplant as a therapeutic modality in treatment of severe lupus. Immune populations and dysfunction in chronic graft vs host disease (CGVHD): In an ongoing natural history protocol (P.I. Steven Pavletic: 04-C-0281), patients who have developed CGVHD following allogeneic transplantation have been evaluated by a multidisciplinary clinical team. The PSC core has supported the study by contributing to the identification of CGVHD biomarkers and the understanding of CGVHD pathogenesis. Four PCS projects have been initiated from this study. First, we have determined that plasma levels of the cytokine BAFF (B cell activating factor of the TNF family) are elevated in CGVHD. This increase is significantly correlated both with elevated plasma levels of Interferon (IFN)-induced inflammatory cytokines and with reduced levels of circulating B cells. We have linked these elevated BAFF levels with increases in transitional B cells, consistent with a reduction in the negative selection process occurring at this maturation stage. Such a change may contribute to the development of the autoimmune symptoms that characterize CGVHD. In a second project, Matin Imanguli, an ETIB clinical research fellow working in the Core, used immunohistochemistry and PCR to determine that severe oral CGVHD of the buccal mucosa was primarily associated with infiltrating T cells expressing T-bet, a transcription factor marking Type I cytokine polarization (Th1/Tc1). Concurrently, we observed increased expression of interferon (IFN)-induced factors including MxA (evidence of Type I IFN), the chemokine MIG (CXCL9) and IL-15 in the affected tissues; these factors support the migration, Th1/Tc1 differentiation and expansion of the T effectors. These data support a model that oral CGVHD results from the interaction of IFN-driven inflammatory processes and Tc1/Th1 differentiated effectors.(Published in 2009, with ongoing studies in erythematous and fibrotic CGVHD of the skin). In a third project exploring IFN-induced processes in CGVHD, we have further correlated these findings in severely affected patients with elevated levels of IFN-induced cytokines and chemokines in the patients plasma. These three studies support the hypothesis that IFN-induced inflammatory processes may underlie many of the systemic processes in CGVHD. These studies formed the basis for an application for a Staff Scientist Innovation Award, funded in January 2010. In a fourth project, we have initiated studies into the role of regulatory T cells (Treg) in CGVHD. Treg cells, which may be involved in controlling autoimmunity, are being assessed in the circulation and in affected tissues. In addition to research on patients in the CGVHD natural history protocol, we support a therapeutic trial for bronchiolitis obliterans, a severe complication of CGVHD (P. I. Ronald Gress and Kirsten Williams: 08-C-0097). We assess leukotriene receptor (LTR) expression in leukocytes and in bronchial lavage cells to assess the role of LTR in progressive fibrosis of lung airways. Cytokine and monokine production following Th2 and Th2/Tc2 therapy based transplant regimens: An ongoing project has assessed lymphokine and monokine production capacity post transplant, focusing on allogeneic transplants incorporating immune therapy with Th2, Th2/Tc2 or Th2.rapa donor-derived cells (P. I. Dan Fowler, 04-C-0055,04-C-0131, 08-C-0088, 07-C-0195). In the early post-transplant period and at regular intervals thereafter, the cytokine and monokine production capacity of peripheral blood cells has been determined by generation of anti CD3/anti CD28-stimulated and bacterial lipopolysaccharide (LPS) stimulated supernatants and by the assessment of the frequencies of cytokine producing T cells by flow cytometry. These assays support these protocols by evaluating the efficacy and durability of the cytokine shift resulting from the infusion of Th2/Tc2 and Th2-rapa cells. Immune reconstitution following lymphodepletion: In several ongoing ETIB clinical trials of allogeneic and autologous stem cell transplantation therapies (04-C-0055, 07-C-0195, 09-C-0210); PIs Daniel Fowler and Michael Bishop) as well as in continuing follow-up of earlier trials (96-C-0104, 99-C-0143, 03-C-0077; P.I.s Ronald Gress, Claude Sportes, Daniel Fowler, Michael Bishop), the process of immune reconstitution has been assessed, focusing on the contributions of thymopoiesis and homeostatic cytokines to the recovery of CD4 and CD8 T cell populations following transplantation. These studies have demonstrated an inverse correlation between circulating plasma levels of IL-7 and CD4 and CD8 T cell populations. Furthermore, in collaboration with T. Fry in POB) they have identified IL-7 levels at 2 weeks post transplant as a potential biomarker predictive of development of acute GVHD. In collaboration with Alan Wayne and Terry Fry (01-C-0125), we have assessed the contributions of donor T cells and thymic-dependent repopulation in the recovery of T cell populations in pediatric transplant recipients. These studies demonstrate the role of T-replete transplants and reduced intensity cytoreductive regimens in producing a rapid repopulation of T cells in young recipients. Finally, when lineage-specific repopulation is important, we work with the ETIB Flow Cytometry Core facility (William Telford) to sort lymphocytes for subset-specific donor chimerism analysis. These assays include ongoing studies on patients treated for myeloid malignancy in the context of MonoMacs congenital immune deficiency (09-C-0096, PI: Dennis Hickstein: repopulation by donor monocyte, T, B and NK cells) and on patients receiving dual donor umbilical cord blood (09-C-0210, PI: Michael Bishop: NK chimerism.

临床前服务核心（PSC）项目是由Etib临床人员实施的移植协议开发的。使用外周血和骨髓以及肿瘤和CGVHD组织活检，我们评估了淋巴细胞亚群，细胞因子含量，T细胞受体库库多样性和胸腺异端活性。所有数据均纳入特定于协议的电子表格，将样品与协议臂和移植时间点联系起来，并可以通过安全的NIH网络访问分支机构。 IL-7给药的影响：与IL-7给药的I期临床试验相关（P. I. Claude Sportes：03-C-0152，与POB中的Crystal Mackall合作），我们确定IL-7显着扩展了天真的和中心的内存CD4和CD8 T细胞，与效应的细胞相比，与CYLER相比，CD4和CD8 T细胞的整体受体相比会增加。这些数据于2009年发布（Sportes等，2009年）。在对该疗法的临床作用的进一步研究中，我们确定IL-7产生了早期B细胞淋巴细胞的短暂扩张，这在外周血中很明显，这是过渡（T1）未成熟B细胞的增加。这些数据被纳入有关2010年IL-7治疗的临床作用的报告中（Sportes等，2010年）。这些研究支持使用IL-7疗法来增加胸腺无能能力有限的患者（例如衰老人群和化学疗法后的免疫缺陷患者）的T细胞数量和曲目。持续研究将重点介绍IL-7与老年患者疫苗反应的作用。严重的全身性狼疮eRythematosus（SLE）患者进行强化淋巴结凝结和自体CD34+干细胞移植后，不对的重建：在正在进行的试验中（P.I. Steven Pavletic 04-C-0095），T和B细胞进行了调节，以评估TIM的效率，并确定了cy的效率，并确定了效率。免疫恢复。这些持续研究有助于自体移植作为严重狼疮治疗的治疗方式的发展。慢性移植与宿主疾病（CGVHD）的免疫种群和功能障碍：在正在进行的自然历史方案（P.I. Steven Pavletic：04-C-0281）中，经过多学科临床团队评估了同种异体移植后发生CGVHD的患者。 PSC核心通过有助于鉴定CGVHD生物标志物和对CGVHD发病机理的理解来支持这项研究。这项研究已经启动了四个PC项目。首先，我们确定CGVHD中细胞因子BAFF的血浆水平（TNF家族的B细胞激活因子）升高。这种增加与干扰素（IFN）诱导的炎症细胞因子的血浆水平升高以及循环B细胞水平降低显着相关。我们已经将这些升高的BAFF水平与过渡B细胞的增加联系起来，这与在此成熟阶段发生的负选择过程的减少一致。这种变化可能有助于表征CGVHD的自身免疫性症状的发展。在第二个项目中，在核心工作的Etib临床研究研究员Matin Imanguli使用免疫组织化学和PCR确定颊粘膜的严重口服CGVHD主要与表达T型T型T-表达T-BET的T细胞（转录因子标记I型细胞因子极化（TH1/TC1））。同时，我们观察到干扰素（IFN）诱导的因子的表达增加，包括MXA（I型IFN的证据），趋化因子MIG（CXCL9）和IL-15在受影响的组织中；这些因素支持迁移，T1/TC1分化和T效应子的扩展。这些数据支持了一个模型，该模型是由IFN驱动炎症过程与TC1/TH1分化效应子的相互作用引起的。在探索IFN引起的CGVHD过程的第三个项目中，我们在严重影响的患者中进一步将这些发现与患者血浆中IFN诱导的细胞因子和趋化因子水平升高相关。这三项研究支持以下假设：IFN诱导的炎症过程可能是CGVHD中许多系统过程的基础。这些研究构成了申请员工科学家创新奖的基础，该奖项于2010年1月资助。在第四个项目中，我们启动了调节性T细胞（TREG）在CGVHD中的作用的研究。可能与控制自身免疫性有关的Treg细胞正在循环和受影响的组织中进行评估。除了对CGVHD自然历史方案中的患者进行研究，我们还支持一项针对细支气管炎的治疗试验，这是CGVHD的严重并发症（P. I. Ronald Gress和Kirsten Williams：08-C-0097）。我们评估白细胞和支气管灌洗细胞中的白三烯受体（LTR）表达，以评估LTR在肺气道进行性纤维化中的作用。 Cytokine and monokine production following Th2 and Th2/Tc2 therapy based transplant regimens: An ongoing project has assessed lymphokine and monokine production capacity post transplant, focusing on allogeneic transplants incorporating immune therapy with Th2, Th2/Tc2 or Th2.rapa donor-derived cells (P. I. Dan Fowler, 04-C-0055,04-C-0131, 08-C-0088，07-C-0195）。在移植后期和此后的定期间隔中，周围血细胞的细胞因子和单因子的产生能力已通过产生抗CD3/抗CD28刺激的抗CD3/抗CD28刺激和细菌脂多糖（LPS）刺激上清液以及通过评估细胞因子产生细胞的频率的频率流动cytry cytry的频率。这些测定方法通过评估由Th2/Tc2和Th2-rapa细胞输注而导致的细胞因子转移的功效和耐用性来支持这些方案。 淋巴结序列后的免疫重建：在几项正在进行的同种异体和自体干细胞移植疗法的ETIB临床试验中（04-C-0055，07-C-0195，09-C-0210）;皮斯·丹尼尔·福勒（Pis Daniel Fowler）和迈克尔·毕晓普（Michael Bishop））以及在继续进行早期试验（96-C-0104，99-C-0143，03-C-0077; P.I.S Ronald Gress，Claude Sportes，Daniel Fowler，Michael Bishop），《免疫恢复的过程》（Themos of Themot and themop cy）的贡献，该过程的贡献是移植后CD4和CD8 T细胞种群。这些研究表明，IL-7和CD4和CD8 T细胞种群的循环血浆水平之间存在逆相关性。此外，与POB中的T. Fry合作，他们在移植后2周确定了IL-7水平，作为潜在的生物标志物预测急性GVHD的发展。与Alan Wayne和Terry Fry（01-C-0125）合作，我们评估了供体T细胞的贡献和胸腺依赖性重生对小儿移植受者T细胞种群恢复的贡献。这些研究表明，T型移植和降低的强度细胞减少方案在产生年轻受体中T细胞的快速重生中的作用。最后，当谱系特异性重新群很重要时，我们与Etib流式细胞仪核心设施（William Telford）合作，对淋巴细胞进行分类以进行亚集特异性供体嵌合分析。 These assays include ongoing studies on patients treated for myeloid malignancy in the context of MonoMacs congenital immune deficiency (09-C-0096, PI: Dennis Hickstein: repopulation by donor monocyte, T, B and NK cells) and on patients receiving dual donor umbilical cord blood (09-C-0210, PI: Michael Bishop: NK chimerism.