Biology of the Immune Response

免疫反应生物学

• 批准号: 7066803
• 负责人: DAVID NELSON
• 金额: --
• 依托单位: DIVISION OF CLINICAL 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/7066803
• 关键词: AIDS; B lymphocyte; CD40 molecule; Wiskott Aldrich syndrome; clinical research; clinical trial phase I; flow cytometry; gene expression; gene therapy; genetic disorder; human subject; human therapy evaluation; hypogammaglobulinemia; immune response; immunopathology chemotherapy; immunopathology therapy; inborn immunodeficiency; lymphocyte proliferation; microarray technology; molecular pathology; monocyte; neoplasm /cancer immunology; patient oriented research; receptor expression; sex linked trait; tumor necrosis factor alpha; yeast two hybrid system

Studies were performed to examine the human immune response in normal individuals and in patients with congenital and acquired immunodeficiency states associated with a high frequency of cancer. We have focused our current studies on patients with the Wiskott-Aldrich syndrome (WAS), X-linked hyper-IgM syndrome, selective IgA deficiency, Common Variable Immune Deficiency (CVID) and X-linked agammaglobulinemia and isolated growth hormone deficiency (XLA/GHD). We continue to study the function of the Wiskott-Aldrich Syndrome Protein (WASP). We have identified patients with spontaneous genetic reversions in the WASP gene and these studies have provided evidence for a selective advantage of gene corrected cells. This observation raises the possibility of a gene correction therapeutic strategy for the Wiskott-Aldrich syndrome. In fact, using such a strategy we have corrected the abnormal structural and functional phenotype of primary T-cells from WAS patients. Further studies on hemotopoetic stem cells are planned. We have also shown that the evaluation of cellular chimerism with regard to WASP expression in WAS patients after bone marrow transplantation can be easily accomplished by flow cytometry. Use of this approach should provide insight into the necessary elements for stem cell correction of the WAS lead to more scientifically-grounded bone marrow treatment strategies for the WAS.Two Phase I clinical trials have recently been completed: 1) recombinant CD40 ligand treatment of patients with X-linked hyper IgM syndrome and 2) BLyS treatment of selective IgA deficiency. Phase II clinical trials are currently being planned for both diseases. Studies have suggested a genetic basis for CVID and selective IgA deficiency. We view these diseases as allelic in nature and the result from a "leaky" failure of immunoglobulin class-switch recombination. To gain possible insight into aberrant gene expression, we are using cDNA microarrays to analyze differential gene expression in stimulated CVID and normal B-cells. We have identified a candidate gene for CVID and selective IgA deficiency. This gene is mutated in approximately 30% of the 100 patient samples already analyzed. We currently have in hand another 300 patient samples for analysis. Knowledge of the candidate gene reveals a pathway of function which is deserving of additional investigations which are currently being planned. X-linked agammaglobulinemia and isolated growth hormone deficiency (XLA/GHD) shares many features with the more common X-linked agammaglobulinemia (XLA) or Bruton's disease caused by mutations in Bruton's tyrosine kinase (Btk). We have shown XLA/GHD to be distinct from XLA since the former patients have no abnormalities in Btk expression including normal catalytic activity. We have shown that patients with XLA/GHD have mutations in a gene called E74-like factor 4(ELF4), or myeloid elf-1-like factor(MEF)-GeneID: 56501. This gene is mutated in all 5 affected male patients, all 3 female disease carriers are heterozygous for the mutation and the mutation was not observed in over 600 normal X-chromosomes examined. Current studies are focusing on the functional consequences of this gene mutation

进行的研究旨在检查正常个体以及患有与高发癌症相关的先天性和获得性免疫缺陷状态的患者的人体免疫反应。我们目前的研究重点是患有 Wiskott-Aldrich 综合征 (WAS)、X 连锁高 IgM 综合征、选择性 IgA 缺乏症、常见变异免疫缺陷 (CVID) 和 X 连锁无丙种球蛋白血症和孤立性生长激素缺乏症 (XLA/GHD) 的患者。我们继续研究 Wiskott-Aldrich 综合征蛋白 (WASP) 的功能。我们已经确定了 WASP 基因自发基因逆转的患者，这些研究为基因校正细胞的选择性优势提供了证据。这一观察结果提出了针对 Wiskott-Aldrich 综合征的基因校正治疗策略的可能性。事实上，利用这样的策略，我们已经纠正了 WAS 患者原代 T 细胞的异常结构和功能表型。计划对造血干细胞进行进一步的研究。我们还表明，通过流式细胞术可以轻松完成骨髓移植后 WAS 患者 WASP 表达的细胞嵌合评估。使用这种方法应该可以深入了解 WAS 干细胞校正的必要要素，从而为 WAS 提供更科学依据的骨髓治疗策略。最近完成了两项 I 期临床试验：1）重组 CD40 配体治疗 X 连锁高 IgM 综合征患者和 2）BLyS 治疗选择性 IgA 缺乏症。目前正在计划针对这两种疾病进行二期临床试验。研究表明 CVID 和选择性 IgA 缺乏症有遗传基础。我们认为这些疾病本质上是等位基因疾病，是免疫球蛋白类别转换重组“泄漏”失败的结果。为了尽可能深入了解异常基因表达，我们使用 cDNA 微阵列来分析受刺激的 CVID 和正常 B 细胞中的差异基因表达。我们已经确定了 CVID 和选择性 IgA 缺乏症的候选基因。在已分析的 100 份患者样本中，大约 30% 的基因发生突变。目前我们手上还有另外 300 份患者样本可供分析。对候选基因的了解揭示了一条功能途径，值得目前正在计划的额外研究。 X 连锁无丙种球蛋白血症和孤立性生长激素缺乏症 (XLA/GHD) 与更常见的 X 连锁无丙种球蛋白血症 (XLA) 或由布鲁顿酪氨酸激酶 (Btk) 突变引起的布鲁顿病具有许多特征。我们已经证明 XLA/GHD 与 XLA 不同，因为前者的 Btk 表达没有异常，包括正常的催化活性。我们已经证明，XLA/GHD 患者的 E74 样因子 4 (ELF4) 或髓样 elf-1 样因子 (MEF) 基因发生突变，基因 ID：56501。该基因在所有 5 名受影响的男性患者中均发生突变，所有 3 名女性疾病携带者均为突变杂合子，并且在检查的 600 多个正常 X 染色体中未观察到该突变。目前的研究重点是该基因突变的功能后果