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目的探讨10例Wiskott-Aldrich综合征(WAS)患儿的临床及分子特点。方法总结10例拟诊WAS患儿临床资料,包括血常规、免疫功能、骨髓常规和扫描电镜检查及临床表型评分。流式细胞术(FCM)检测10例患儿外周血单个核细胞(PBMC)中WAS蛋白(WASP)表达。PCR扩增WASP基因序列并直接双向测序分析9例患儿及其亲属突变情况。结果本组均男性,多以自幼大便带血丝及皮肤瘀点瘀斑起病。均有血小板减少伴小血小板,湿疹和免疫缺陷表现。临床表型评分3例评5分,3例评4分,4例评3分。具有阳性家族史患儿临床诊断年龄明显早于无家族史者。多数患儿IgA(8/9)、IgE(8/9)和IgG(7/9)升高,除6例CD4~+T比例下降(6/9),其余患儿淋巴细胞亚群分类正常。1例淋巴细胞增殖功能降低(1/3)。骨髓常规缺乏特征性改变。5例患儿淋巴细胞扫描电镜(SEM)均可见典型微绒毛异常。10例患儿WASP均为阴性,9例行WASP基因分析发现7种不同突变,3例为新型突变(168 C>A,T45 K;747-748 del T,I 238 Fs X260;253 Ins A,C73X)。8例位于编码区,1例位于内含子。4例突变位于EVH1区,2例BR区,1例VCA区,1例GBD区。突变类型包括无义突变4例(2例155 C>T,R 41 X;2例665 C>T,R 211 X),错义突变2例(168 C>A,T45 K~*;1487G>A,D485N/5’剪切),缺失突变1例(747-748 del T,I 238 Fs X260~*),拼接位点突变1例(Ivs9+2 T>C),插入突变1例(253 Ins A,C73X~*)。8例患儿母亲为突变携带者。除预防性抗生素及静注丙球(IVIG),3例接受干细胞移植,移植后2例正常表达WASP,1例死于巨细胞病毒(CMV)所致的肺间质性病变。结论早发持续血小板减少伴血小板体积减少的男性患儿应怀疑WAS及相关疾病、WAS蛋白检测和基因分析是明确诊断的重要手段。基因分析还有利于发现携带者和进行遗传咨询。干细胞移植是目前根治WAS,尤其是重症患儿最有效的方法。
Objective To investigate the clinical and molecular characteristics of 10 children with Wiskott-Aldrich syndrome (WAS). Methods The clinical data of 10 children with suspected WAS were summarized, including blood routine, immune function, routine bone marrow and scanning electron microscopy and clinical phenotype score. Flow cytometry (FCM) was used to detect WAS protein expression in peripheral blood mononuclear cells (PBMCs) of 10 children. PCR amplification of WASP gene sequence and direct bi-directional sequencing analysis of 9 cases of children and their relatives mutation. Results This group of men, mostly with stool of blood stasis and skin petechiae onset. Thrombocytopenia with small platelets, eczema and immune deficiency performance. Clinical phenotype score 3 cases rated 5 points, 3 cases rated 4 points, 4 cases rated 3 points. Children with a positive family history of clinical diagnosis of children was significantly earlier than those without family history. The majority of children with IgA (8/9), IgE (8/9), and IgG (7/9) were elevated except for 6 cases of CD4 ~ + T (6/9), and the remaining children had normal lymphocyte subsets . One case of lymphocyte proliferation decreased (1/3). Bone marrow routine lack of characteristic changes. 5 cases of children with lymphocyte scanning electron microscopy (SEM) showed typical microvilli abnormalities. Ten cases of children with WASP were negative, nine cases of WASP gene analysis found seven different mutations, three cases of new mutations (168 C> A, T45 K; 747-748 del T, I 238 Fs X260; 253 Ins A, C73X). Eight were in the coding region and one was in the intron. Four mutations were located in EVH1, two in BR, one in VCA and one in GBD. There were 4 cases of nonsense mutations (2 cases of 155 C> T, R 41 X; 2 cases of 665 C> T, R 211 X) and 2 missense mutations (168 C> A, T45 K ~ *; 1487G> A, D485N / 5 ’cut), 1 case of deletion mutation (747-748 del T, I 238 Fs X260 ~ *), 1 case of splicing site mutation (Ivs9 + 2 T> Ins A, C73X ~ *). Eight mothers were mutants. In addition to prophylactic antibiotics and IVIG, 3 received stem cell transplantation, 2 received normal WASP and 1 died of interstitial lung disease due to cytomegalovirus (CMV). Conclusions WAS and related diseases should be suspected in the male patients with early thrombocytopenia and thrombocytopenia. WAS protein detection and gene analysis are important means to confirm the diagnosis. Genetic analysis also facilitates the discovery of carriers and genetic counseling. Stem cell transplantation is the most effective method to cure WAS, especially severe children.