Function of the gene of Wiskott-Aldrich syndrome

1)Wiskott-Aldrich综合征WASP基因突变5例，其中1、2例外显子1碱基(G)缺失，3例外显子2 TCA*TAA改变，4例10外显子CGA*TGA改变，5例10外显子1碱基缺失(C)。例1、例2用聚合酶链式反应-单链构象多态性方法检测WASC携带者状态，例4用Xhol法检测，例5用直接测序法检测WASPmRNAWASPmRNA的表达。患者外周血中未检测到WASPmRNA。4)抗WASP多克隆和单克隆抗体的产生及免疫印迹分析检测WASP多肽的多克隆和克隆(5A5)抗体被证明是针对WASP的多克隆和单克隆(5A5)抗体，使用这些抗体，WASP在患者的单个核细胞中不表达。5)用5A5单抗流式细胞术检测WASP在患者的淋巴母细胞系细胞质中呈阴性。6)接受骨髓移植并继发EB病毒相关性B淋巴细胞增生性疾病的IS患者成功接受供者淋巴细胞输注治疗。

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.