STUDIES OF NATURALLY OCCURRING DEFECTS IN RESISTANCE TO INFECTION

天然存在的抗感染缺陷的研究

• 批准号: 7379391
• 负责人: HANS D OCHS
• 金额: $ 0.58万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7379391
• 关键词: STUDIES; NATURALLY; OCCURRING; DEFECTS; RESISTANCE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Primary immunodeficiency diseases are unique human models to study the molecular events that ultimately lead to an efficient immune response. With the support of this grant, our group has participated in the identification of genes involved in X-linked immunodeficiency diseases and in the functional analysis of the products of these genes. The understanding of the molecular basis for immune disorders has contributed greatly to the concept of receptor-ligand interaction, and the principles of lymphocyte activation, differentiation, signal transduction and immunoglobulin isotype switching. Three specific aims will be addressed. (1) Elucidation of the gene responsible for the Wiskott-Aldrich syndrome (WAS) and its product, the WAS protein. We have been part of a team that recently isolated the gene mutated in patients with WAS, a disorder in which most hematopoietic cell lineages are affected. Our collection of B and T cell lines derived from members of over 50 affected families will permit a representative mutation analysis of WAS and will answer the question of whether a correlation exists between clinical phenotype and type of mutation. Based on the amino acid composition derived from the known cDNA sequence, we have outlined strategies to analyze the nature of the WAS protein. We will focus on WAS gene expression and its control, its distribution within the cell, the physical and functional properties of the WAS protein, will identify other proteins that interact with WASP. (2) Explore the role of defective transcriptional activation of T cells in patients with common variable immunodeficiency (CVI). We have identified a subgroup of CVI patients who have in common a broad defect of T cell activation, suggesting that the transcription of multiple lymphokines and membrane bound activation molecules may be abnormal. Using gelshift assays, we will determine if the defect is due to abnormal binding of NF-AT or other nuclear factors related to the IL-2 gene. If decreased binding is demonstrated, we will search for defective production of proteins of the NF-AT complex and analyze its distribution in the cytoplasm and in the nuclear fraction, and its phosphorylation state. (3) Explore the feasibility of gene therapy for patients with X-linked immunodeficiencies. We have initiated collaboration with several investigators involved in the design of viral vectors that efficiently transfer the candidate genes into target cells, e.g., T and B cell lines or stem cells derived from patients.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。原发性免疫缺陷疾病是研究最终导致有效免疫反应的分子事件的独特人类模型。在这笔赠款的支持下，我们的小组参与了鉴定参与X连锁免疫缺陷疾病的基因以及对这些基因产物的功能分析。对免疫疾病的分子基础的理解对受体配体相互作用的概念以及淋巴细胞激活，分化，信号转导和免疫球蛋白同种型切换的原理产生了巨大贡献。将解决三个具体目标。 （1）阐明负责Wiskott-Aldrich综合征（WAS）及其产物的基因。 我们一直是一个最近分离出在患者中突变的基因的团队的一部分，这种疾病大多数造血细胞谱系受到影响。我们收集的B和T细胞系从50多个受影响家庭的成员中得出的B和T细胞系将允许对WAS进行代表性突变分析，并将回答有关临床表型和突变类型之间是否存在相关性的问题。基于源自已知的cDNA序列的氨基酸组成，我们概述了分析IS蛋白质性质的策略。我们将重点关注的是基因表达及其对照，其在细胞中的分布，是IS蛋白质的物理和功能特性，将识别与WASP相互作用的其他蛋白质。 （2）探讨T细胞在常见可变免疫缺陷（CVI）患者中T细胞有缺陷的转录激活的作用。我们已经确定了一个CVI患者的亚组，这些患者共同存在T细胞激活的广泛缺陷，这表明多种淋巴细胞和膜结合活化分子的转录可能是异常的。使用GelShift分析，我们将确定缺陷是由于NF-AT或与IL-2基因相关的其他核因子的异常结合所致。如果证明结合的减少，我们将搜索NF-AT复合物的蛋白质的有缺陷的产生，并分析其在细胞质和核分裂中的分布及其磷酸化状态。 （3）探讨基因治疗对X连锁免疫缺陷患者的可行性。我们已经与参与设计病毒载体设计的几位研究人员合作，这些研究人员有效地将候选基因转移到靶细胞中，例如T和B细胞系或来自患者的干细胞。