Function of the gene of Wiskott-Aldrich syndrome

Wiskott-Aldrich 综合征基因的功能

基本信息

批准号：
08457219
负责人：
TSUCHIYA Shigeru
金额：
$ 0.9万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

1) Mutation of WASP gene in Wiskott-Aldrich syndrome (WAS)Mutation of WASP gene was found in 5 cases of WAS.One base (G) deletion in the exon 1 in cases 1 and 2, TCA*TAA change in the exon 2 in case 3, CGA*TGA change in the exon 10 in case 4 and one base deletion (C) in the exon 10 in case 5 were found. These mutations were suggested to give rise to truncated WASP of 63,63,53,320 and 444 amino acids, respectively.2) Detection of carrier status of WASCarriers in family members were detected by PCR-SSCP in case 1 and 2, by Xhol digestion in case 4 and by direct sequencing of PCR product in case 5.3) Expression of WASPmRNAWASPmRNA was expressed in mononuclear cells and granulocytes from healthy volunteers by Northern blot analysis. WASPmRNA was not detected in those from the patients. The mRNA was found in human leukemia sell lines in any origin, but not in human fibroblasts.4) Production of polyclonal and monoclonal antibodies to WASP and detection of WASP by Western blot analysisPolyclonal and monoclonal (5A5) antibodies were made to WASP peptides and proved to be specific to WASP.Using these antibodies WASP was shown not to express in mononuclear cells of the patients.5) Flow cytometric detection of cytoplasmic WASP using the 5A5 monoclonal antibodyWASP was clearly negative in cytoplasm of the lymphoblastoid cell lines from the patients as compared in that from normal volunteers. However, strong nonspecific binding interfered the detection of WASP in monocytes, lymphocytes and granulocytes from the patients.6) Case report of WAS who received bone marrow transplantation and subsequent EBV related B lymphoproliferative diseaseThe patient was treated by donor lymphocyte transfusion successfully.

1）在Wiskott-Aldrich综合征中WASP基因突变（WAS）在5例WAS. WAS的突变（WAS）突变是在外显子1和2中的外显子1和2中的1和2中，TCA*TAA在案例3中的TCA*TAA变化，cga*tga在exon 10中的CGA*TGA在exon 10中的CGA*TGA在4和一个基本deletion（c）中（c）中的exon（c）中有5个exon 10中的10 exon中的10个。建议这些突变产生分别导致63,63,53,320和44444种氨基酸的截短的黄蜂。2）在案例4中通过Xhol Digestion在案例4和PCR的直接测序中，在案例4和PCR中，PCR-SSCP在案例1和2中检测到家族成员中的载体状态的检测。通过Northern印迹分析来自健康志愿者的粒细胞。在患者的患者中未检测到黄蜂。在人白血病中发现了mRNA的任何来源，但在人的成纤维细胞中却没有发现。4）生产对WASP的多克隆和单克隆抗体的产生，并通过蛋白质印迹分析蛋白质分析和单克隆分析和单克隆分析（5a5）（5A5）（5AA5）对WASP的抗体进行了抗体，并在这些抗体中表现出了这些抗体。 5）5）使用5A5单克隆抗体抗体对细胞质WASP的流式细胞仪检测显然在患者的淋巴细胞细胞系的细胞质中为阴性，而正常志愿者的淋巴细胞细胞系则显然为。然而，强的非特异性结合干扰了患者的单核细胞，淋巴细胞和粒细胞中的黄蜂的检测。6）供体lymphopyte Transfisical的供体疾病的病例报告。