CORE--PATIENT MUTATION ANALYSIS

患者突变分析核心是血液处理，组织培养 和突变分析核心 在这里，我们收集并处理 WAS和XLT患者的外周血，包括定量和 血细胞亚群的表型分型，单核细胞的分离， 淋巴细胞，冷冻保存，以及生成，维持和 EBV转化的B细胞系的表征，其将用于 项目1和项目2的调查。 核心有助于诊断 在新转诊患者中进行血小板测定， 缺乏家族史，女性亲属杂合性分析。 核心鉴定诊断患者的WAS基因突变， 分离DNA并扩增外显子区域，分析扩增的区域， 随后进行单链构象多态性分析（SCCP）， 异常区域的测序。 核心表征和量化 通过以下组合在患者细胞中表达突变的WASP蛋白： 蛋白质印迹和磷酸成像。 核心将决定 加入蛋白酶抑制剂，一种潜在的治疗方法， 提高部分患者培养细胞中WASP蛋白水平。 最后，核心维护患者血液样本和细胞的数据库 线以及临床和分子信息参与 患者和已发表WAS缺陷的单独数据库。

The Patient Mutation Analysis Core, is a blood processing, tissue culture and mutation analysis core. In this facility, we collect and process peripheral blood of WAS and XLT patients including quantifying and phenotyping blood cell subpopulations, isolation of mononuclear cells and lymphocytes, cryopreservation, and generation, maintenance and characterization of EBV-transformed B-cell lines that will be used in investigations in Projects 1 and 2. The Core contributes to diagnosis of WAS in newly referred patients by performing platelet sizing and, where family history is lacking, heterozygosity analysis of female relatives. The Core identifies the WAS gene mutation of diagnosed patients by isolating DNA and amplifying exon regions, analyzing amplified regions by single chain conformation polymorphism analysis (SCCP) followed be sequencing of the abnormal region. The Core characterizes and quantifies the mutated WASP protein expressed in patient cells by a combination of Western blotting and Phosphor-Imaging. The Core will determine whether addition of protease inhibitors, a potential therapeutic treatment, will increase the level of WASP protein in cultured cells of some patients. Finally the Core maintains a database of patient blood samples and cell lines as well as clinical and molecular information on participating patients and a separate database of published WAS defects.