Function of the gene of Wiskott-Aldrich syndrome

1)Wiskott-Aldrich综合征（WAS）的WASP基因突变5例WAS中发现WASP基因突变，例1和例2的外显子1有一个碱基（G）缺失，例3的外显子2有TCA*TAA改变，病例4中外显子10的CGA*TGA变化和一个碱基缺失（C）病例5的WASP基因外显子10中发现了10个突变，这些突变可能导致WASP基因的63、63、53、320和444个氨基酸的截短，2）病例1、2采用PCR-SSCP方法检测其家庭成员WASC携带者，在情况4中通过XhoI消化和在情况5.3中通过PCR产物的直接测序）北方印迹法检测健康人单核细胞和粒细胞中WASPmRNA的表达。WASPmRNA在患者组织中未检测到。在任何来源的人白血病细胞系中发现了这种mRNA，4）WASP多克隆抗体和单克隆抗体的制备及WASP蛋白的Western blot检测制备了WASP多肽的多克隆抗体和单克隆抗体（5A 5），并证明其对WASP具有特异性，用这些抗体检测WASP蛋白在患者的单核细胞中不表达。用5A 5单克隆抗体流式细胞术检测胞浆WASP，与正常人相比，患者淋巴母细胞系胞浆WASP明显阴性。6）骨髓移植后发生EB病毒相关B淋巴细胞增生性疾病的WASP患者1例。

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.