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: 7733342
• 负责人: Frances Hakim
• 金额: $ 70.14万
• 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/7733342
• 关键词: Acute; Acute Graft Versus Host Disease; Aging; Allogenic; Apoptotic; Autoimmunity; Autologous; Autologous Stem Cell Transplantation; B-Lymphocytes; Biological Assay; Biological Markers; Biopsy; Blood Cells; Bronchial Lavages; Bronchiolitis Obliterans; CD34 gene; CD4 Positive T Lymphocytes; CD8B1 gene; Cells; Chronic; Clinical; Clinical Research; Clinical Trials; Collaborations; Competence; Complication; Confocal Microscopy; Cytotoxic T-Lymphocytes; DNA Sequence Rearrangement; Data; Databases; Development; Effector Cell; End Point; Enzyme-Linked Immunosorbent Assay; Excision; Family; Fibrosis; Flow Cytometry; Frequencies; Functional disorder; Generations; Goals; Hematopoietic Stem Cell Transplantation; Homeostasis; Homologous Transplantation; Immune; Immune system; Immunohistochemistry; Immunotherapy; Infection; Inflammatory; Infusion procedures; Institutes; Interleukin-4; Interleukin-7; Leukocytes; Leukotriene Receptor; Leukotrienes; Link; Lipopolysaccharides; Lung diseases; Lupus; Lymphocyte; Lymphocyte Depletion; Lymphocyte Subset; Lymphokines; Marrow; Memory; Modality; Monitor; Monoclonal Antibody HuM291; Monokines; Morbidity - disease rate; Muromonab-CD3; Natural History; Natural regeneration; Oral; Pathogenesis; Pathway interactions; Patients; Peripheral; Peripheral Blood Lymphocyte; Pharmaceutical Preparations; Phase I Clinical Trials; Plasma; Polymerase Chain Reaction; Population; Predisposition; Process; Production; Protocols documentation; Recovery; Relative (related person); Sampling; Secure; Services; Staining method; Stains; Stem cell transplant; Support of Research; Systemic Lupus Erythematosus; T memory cell; T-Cell Receptor; T-Lymphocyte; TALL-1 protein; TNF gene; Testing; Therapeutic; Therapy Clinical Trials; Time; Tissues; Transplantation; Transplantation Immunology; Treatment Protocols; United States National Institutes of Health; Upper arm; Vaccines; Work; base; chemotherapy; chronic graft versus host disease; cytokine; follow-up; graft vs host disease; immune function; mortality; multidisciplinary; peripheral blood; pre-clinical; reconstitution; response; tumor

The specific projects in which the Preclinical Service has become involved were initiated by transplantation protocols implemented by the clinical staff of Experimental Transplantation and Immunology Branch. We have cryopreserved patient lymphocytes from peripheral blood and marrow, tumor and CGVHD tissue biopsies, as well as plasma, and created a searchable database linking samples to protocol and transplant time points. We have then examined these samples for lymphocyte subsets, cytokine content and 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), the effects of this therapy were assessed by multi-parameter flow cytometry, quantitation of T cell receptor rearrangement excision circles (TREC), and spectratyping of T cell receptor repertoire diversity. In this trial IL-7 therapy was found to significantly increase peripheral blood CD4 and CD8 T cell populations. We further analyzed patient samples to investigate mechanism of action. We determined that IL-7 treatment produced a marked increase in the relative proportion of nave and central memory T cells, at the expense of effector T cells, especially the CD45RA+CD27- effector CD8 populations. IL-7 treatment produced an increase in the percentage of peripheral blood T cells in cycle (ki-67+) and increased cellular levels of the anti-apoptotic factor bcl-2; the differential effects of IL-7 on T subset expansion were associated with the persistence of the proliferation and apoptotic protection in nave cells, as compared to effector cells. Furthermore, we determined that these shifts were associated with a dilution of TREC in the most nave CD31+CD45RA+ population of CD4 cells, consistent with a primary mechanism for IL-7 therapy of thymic-independent expansion of peripheral T cell populations. Finally, IL-7 was shown to produce a significant increase in the diversity of the overall T cell receptor repertoire. These assays contributed to an understanding of the mechanism of IL-7 effects. Furthermore they supported the use of IL-7 therapy to enhance T cell repertoire and immune functional competence in patients with limited thymopoietic capacity, such as in aging populations and in patients with immune deficits after chemotherapy. 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 by multi-parameter flow cytometry, TREC quantitation and plasma cytokine assays to evaluate recovery of thymopoiesis, reconstitution of naive, memory and effector T cell populations and normalization of immune function in patients with severe autoimmunity. These studies assessed the efficacy of the cytoreductive regimen in depleting T and B cells and the timecourse of immune recovery. These studies contribute to development of autologous transplant as a therapeutic modality in treatment of severe lupus and to an understanding of immune system homeostasis and regeneration in the context of autoimmunity. Immune populations and dysfunction in chronic graft vs host disease: In an ongoing trial (P.I. Steven Pavletic 04-C-0281), patients who have developed chronic GVHD following allogeneic transplantation are evaluated at NIH in a multi-institute natural history trial. The PCS core has evaluated T and B cell immune reconstitution and function in these patients by multi-parameter flow cytometry, TREC and cytokine assays, and by PCR analysis and confocal microscopy of tissue biopsies stained by fluorescent immunohistochemistry. These assays support the ongoing multidisciplinary natural history study of chronic GVHD, by contributing to the identification of biomarkers and to an understanding of chronic GVHD pathogenesis. We have determined that elevated levels of B cell activating factor of the TNF family (BAFF)in chronic GVHD are significantly correlated both with elevated plasma levels of inflammatory cytokines and with reduced levels of circulating B cells. Furthermore we have linked these elevated BAFF levels with persistence of transitional B cells, consistent with a reduction in negative selection of autoreactive cells. In addition, studies of peripheral blood populations and cytokines have been correlated with immunohistochemical and PCR analyses (performed by an ETIB clinical research fellow, M. Imanguli, working in the Core) of oral biopsy tissues from patients in the chronic GVHD protocol. These studies have demonstrated the involvement of cytotoxic T cell effectors in oral CGVHD. Finally, we support a therapeutic trial for bronchiolitis obliterans, a severe complication of chronic GVHD (P. I. Ronald Gress and Kirsten Williams 08-C-0097). This trial tests the efficacy of a drug blocking the leukotriene pathway to inhibit the progression of fibrosis in this obstructive lung disorder. The core provides multi-parameter flow cytometric assessment of leukotriene receptor levels in peripheral blood leukocytes and in bronchial lavage cells. 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). 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 cells by flow cytometry. These assays support branch transplant protocols by assessing the use of monokine supernatants and frequency of monokine producing cells as a potential biomarkers predicting acute GVHD. Furthermore they evaluate 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 and 07-C-0195; PIs Daniel Fowler and Michael Bishop) as well as in continuing follow-up of patients treated in earlier trials (96-C-0104, 99-C-0143, 03-C-0077; P.I.s Claude Sportes, Daniel Fowler, Michael Bishop), the process of immune reconstitution has been assessed by multiparameter flow cytometry, functional assays, quantitation of TREC, spectratyping analyses of repertoire diversity and ELISA assays of plasma cytokines. These studies have assessed the contributions of thymopoiesis and homeostatic cytokines in the recovery of CD4 and CD8 T cell populations following autologous stem cell transplantation. These data now are used as a basis of comparison in patient [summary truncated at 7800 characters]

临床前服务涉及的特定项目是由实验移植和免疫学分支的临床人员实施的移植方案启动的。我们有来自外周血和骨髓，肿瘤和CGVHD组织活检的冷冻患者淋巴细胞以及血浆，并创建了一个可搜索的数据库，将样品连接到方案和移植时间点。然后，我们检查了这些样品的淋巴细胞亚群，细胞因子含量和T细胞受体库的多样性和胸腺无质活性。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给药的效果：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），通过多参数流式细胞仪，T细胞受体重新培训圆圈（TRECERECERTINE）和SPEMETRARTINES和SPEMETRATYPRATYPRATYPRATYPRATYPRATYPRATYPRATYPRATYPRATYPRATISPINGIPSING和SPEMETRATYPRATYPRATYPRATYPRATYPRATYPRATYPRATISPINGIPSING，通过多参数流式细胞术，多参数流式细胞仪进行了效果。在这项试验中，发现IL-7治疗可显着增加外周血CD4和CD8 T细胞群体。我们进一步分析了患者样本以研究作用机理。我们确定IL-7处理产生了中殿和中央记忆T细胞相对比例的显着增加，牺牲了效应T细胞，尤其是CD45RA+CD27-效应CD8种群。 IL-7治疗在循环（KI-67+）中产生的外周血T细胞百分比增加，抗凋亡因子Bcl-2的细胞水平增加。与效应细胞相比，IL-7对T子集膨胀的差异作用与中殿细胞的增殖和凋亡保护的持久性有关。 此外，我们确定这些转移与CD31+ CD45RA+ CD4细胞种群中TREC的稀释有关，这与IL-7治疗胸腺独立于外周T细胞种群扩张的主要机制一致。最后，证明IL-7会显着增加T细胞受体库的多样性。这些测定有助于理解IL-7效应的机制。 此外，他们还支持使用IL-7治疗来增强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）的患者的T细胞库和免疫功能能力。严重的全身性红斑狼疮（SLE）患者（SLE）患者的强化淋巴结和自体CD34+干细胞移植后，免疫重建。在正在进行的试验中（P.I. Steven Pavletic 04-C-0095），通过多参数流式细胞术，TREC定量和血浆细胞因子分析来监测T和B细胞的重构，以评估胸腺甲状腺素的恢复，幼稚和效应T细胞群体的重新构造，Nemive和Hembore ticle群体的重新构成，并在免疫范围内进行了自动化的患者。这些研究评估了细胞补充方案在耗尽T和B细胞中的功效以及免疫恢复的时间。这些研究有助于自体移植作为治疗严重狼疮的治疗方式的发展，并在自身免疫性的背景下了解免疫系统稳态和再生。慢性移植与宿主病的免疫种群和功能障碍：在正在进行的试验中（P.I.史蒂文·帕维尔特式04-C-0281），在一项多基础自然历史试验中，在NIH评估了在同种异体移植后出现慢性GVHD的患者。 PCS核心通过多参数流式细胞仪，TREC和细胞因子测定法，以及通过荧光免疫组化染色的组织活检，通过多参数流式细胞仪，TREC和细胞因子测定评估了T和B细胞免疫重建和功能。这些测定法支持了慢性GVHD的持续多学科自然史研究，通过有助于鉴定生物标志物和对慢性GVHD发病机理的理解。我们已经确定，慢性GVHD中TNF家族（BAFF）的B细胞激活因子水平升高与血浆炎性细胞因子水平升高以及循环B细胞水平降低显着相关。此外，我们将这些升高的BAFF水平与过渡B细胞的持久性联系在一起，这与自动反应性细胞的负选择相一致。此外，对外周血群体和细胞因子的研究与免疫组织化学和PCR分析（由ETIB临床研究研究员M. imanguli，在核心工作）进行了慢性GVHD方案中患者的口腔活检组织的研究。这些研究表明，细胞毒性T细胞效应子参与口服CGVHD。最后，我们支持一项针对细支气管炎的治疗试验，这是慢性GVHD的严重并发症（P. I. Ronald Gress和Kirsten Williams 08-C-0097）。该试验测试了阻塞白三烯途径抑制这种阻塞性肺疾病中纤维化进展的药物的功效。该核心提供了周围血清细胞和支气管灌洗细胞中白细胞中白三烯受体水平的多参数流式细胞术评估。基于TH2和TH2/TC2治疗后的细胞因子和单因子产生基于移植方案：一个正在进行的项目已经评估了移植后的淋巴因子和单因子生产能力，重点是将免疫治疗与TH2，TH2/TC2或TH2.RAPA供体供体细胞（P. I. Dan Fowler，04-C-cy）纳入同种异体疗法，04-C-c-04-c-c-04-c-c- 04-c-cy。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产生能力已通过产生抗CD3/抗CD28刺激的抗CD3/抗CD28刺激性和细菌脂多糖（LPS）刺激上清液以及通过评估由流动细胞的频率的频率来确定。这些测定方法通过评估单算上清液的使用和单算细胞的频率作为预测急性GVHD的潜在生物标志物来支持分支移植方案。 此外，他们评估了由Th2/Tc2和Th2-Rapa细胞输注而导致的细胞因子偏移的疗效和耐用性。淋巴结序列后的免疫结构：在正在进行的几项同种异体和自体干细胞移植疗法（04-C-0055和07-C-0195; PIS Daniel Fowler和Michael Bishop）的临床试验中P.I.S Claude Sportes，Daniel Fowler，Michael Bishop），通过多参数流式细胞仪，功能测定，TREC的定量，谱分析分析的库库多样性和ELISA测定的谱分析，对免疫重建的过程进行了评估。这些研究评估了自体干细胞移植后CD4和CD8 T细胞种群恢复的胸腺波素和稳态细胞因子的贡献。 这些数据现在被用作患者比较的基础[摘要以7800个字符截断]