Wiskott-Aldrich综合征高危儿七例产前诊断

目的 探讨Wiskott-Aldrich综合征(WAS)高危孕妇羊水脱落细胞基因分析及脐静脉穿刺脐血WAS蛋白(WASP)检测在WAS高危儿产前诊断中的意义.方法 2008年至2010年我院经WASP流式检测和基因分析明确诊断的7例WAS患儿.详细记录先证者病史,进行家族相关亲属基因检测,建立7个WAS家系图谱.2008年至2011年对7个家系中携带异常基因的7个高危孕妇于孕18 ～ 20周经羊膜腔穿刺抽取羊水.其中部分羊水提取细胞DNA,经PCR扩增WAS基因.PCR产物进行双向序列重复测定.其中1例高危孕妇孕28周采集脐带血,采用流式细胞术检测WASP.培养羊水中胎儿脱落细胞,采用原位制片及G带染色技术进行染色体核型分析.产后采集高危儿外周血进行基因分析和WASP检测验证产前诊断结果.结果 7例WAS高危儿羊膜腔穿刺均成功,羊水细胞培养成功率100％.WAS基因和染色体核型分析结果显示1例正常男性胎儿,4例正常女性胎儿,2例为女性异常基因携带者.1例脐带血流式细胞术检测WASP显示正常表达.7例高危儿均顺利出生,产后WASP和基因分析结果均正常,与产前结果相同.结论 羊水脱落细胞核型、基因分析与脐带血WASP检测可为WAS高危孕妇提供可靠的产前诊断服务.     

单核细胞增多为特征的湿疹-血小板减少伴免疫缺陷综合征1例并文献复习

为了探讨湿疹-血小板减少伴免疫缺陷综合征（WAS）的特殊表型。回顾性分析2017年3月重庆医科大学附属儿童医院风湿免疫科收治的1例以单核细胞增多为特征的WAS病例的临床资料及实验室检查结果,并复习相关文献。结果发现,患儿出生后以血小板减少及贫血为主要表现,伴肝脾肿大及间断血便,血常规提示白细胞、中性粒细胞及单核细胞增高,血红蛋白及血小板降低,外周血及骨髓原始幼稚细胞不高,考虑幼年型粒单核细胞白血病（JMML）可能。JMML相关基因和染色体核型未见异常。由于WAS蛋白（WASP）明显表达减少,WAS基因存在一处半合子突变（c.151G>T,p.V51F）,最终确诊为WAS。WAS临床表型变异大。对于早发血小板减少的男婴,建议检测WAS蛋白和WAS基因进行筛查。     

Wiskott-Aldrich 综合征1 例临床及基因分析

目的探讨Wiskott-Aldrich综合征(WAS)的WAS基因突变的特点。方法回顾分析1例WAS患儿的临床资料,以及利用PCR测序方法检测WAS基因全部外显子及侧翼序列。结果 5个月男性患儿,发现血小板减少入院,既往湿疹时间长且反复感染;CD8+及CD4+T淋巴细胞升高,CD 19+B淋巴细胞正常;骨髓细胞学提示巨核细胞成熟障碍。WAS基因检测发现存在C.880 AG(p.Ile294Val)突变,患儿父母均未见突变。利用Polyphen2软件及SIFT软件预测该位点为致病性突变,不同物种间序列保守性分析发现该位点保守;结构预测分析发现该位点突变可能会影响正常的蛋白结构,该突变国内外未见报道。结论基因检测可早期诊断WAS,新发现C.880 AG(p.Ile294Val)突变。     

Wiskott-Aldrich综合征患儿异基因造血干细胞移植后EB病毒相关淋巴组织增殖性疾病1例报告并文献复习

目的 探讨儿童Wiskott-Aldrich综合征(WAS)异基因造血干细胞移植术(allo-HSCT)后EB病毒(EBV)相关淋巴组织增殖性疾病(PTLD)的治疗.方法 回顾分析1例行allo-HSCT的WAS患儿的临床资料,并检索复习相关文献.结果 12岁男性WAS患儿,获骨髓库人类白细胞抗原(HLA)配型10/1...     

Wiskott-Aldrich综合征临床和遗传学诊断:附9例报告

目的探讨Wiskott-Aldrich综合征(WAS)的临床和基因诊断价值。方法对9例WAS患儿作临床表型评分和血常规、免疫学、骨髓常规、扫描电镜检查,并用PCR直接测序法分析4例患儿及其母亲基因组DNA中WAS蛋白(WASP)基因序列。结果9例患儿均为典型WAS,评4分,家庭史阳性者诊断年龄早于家族史阴性者;免疫学结果差异大;骨髓常规易误诊“特发性血小板减少性紫癜”,5例WAS患儿淋巴细胞、血小板扫描电镜有典型异常,其中4例存在WASP基因突变:缺失突变2例(984delC,P317fsX444),无义突变1例(1388G>T,E452X),拼接错误1例(IVIS 1G>T)。结论对怀疑WAS患儿应仔细询问家族史,临床评分,作扫描电镜检查可提高WAS的临床诊断准确率;WASP基因分析可确诊患者,发现携带者,为干细胞移植治疗患者提供依据。     

造血干细胞移植治疗湿疹血小板减少伴免疫缺陷综合征的临床研究——单中心11年回顾性分析

目的探讨异基因造血干细胞移植(allo-HSCT)治疗湿疹血小板减少伴免疫缺陷综合征(WAS)的疗效、早期并发症、预后影响因素及脐血作为WAS移植供体选择的可行性。方法对上海儿童医学中心2006年11月-2018年9月接受allo-HSCT的29例确诊WAS患儿的临床资料进行回顾性分析。结果 29例WAS患儿接受allo-HSCT治疗,其中13例接受脐血(UCB)移植,16例接受外周血干细胞(PBSC)移植,其中同胞全相合供体(MSD)移植2例,全相合无关供体(MUD)移植8例,不全相合无关供体(MMUD)4例,亲缘不全相合供体(MMSD)移植2例。经环磷酰胺+白消安+抗人胸腺球蛋白清髓性预处理后,所有患儿均获得完全植入。总体2年无病生存率为93%(27/29)。UCB和PBSC移植中性粒细胞和血小板植入中位时间分别为12d vs.11d(P=0.215)和47d vs.26d(P=0.133)。69%患者发生aGVHD,均为2～3度皮肤GVHD,UCB移植患者与PBSC移植患者相比aGVHD发生程度及持续时间无差异(P值分别为0.445和0.345)。不全相合较全相合供体移植aGVHD持续时间更长(P=0.001),但两者间aGVHD发生程度及频率差异无显著性(P=0.875)。导管相关血行感染主要发生于围移植期,发生率26.1%。结论 allo-HSCT治疗WAS总体预后好,脐血是WAS患儿合适的造血干细胞来源。     

两种新型Wiskott-Aldrich综合征蛋白基因突变的鉴定

目的 明确3例Wiskott-Aldrich综合征(WAS)患儿WAS蛋白(WASP)基因突变的类型。方法 根据典型临床表现(血小板减少、湿疹、反复感染),及淋巴细胞和血小板扫描电镜改变,采用PCR直接测序法,对3例疑为WAS的患儿及其母亲的WASP基因进行序列分析。结果 以正义、反义引物扩增的PCR产物分别测序,发现两种新型WASP基因突变：2例WAS孪生兄弟WASP基因第10外显子,第984位核苷酸C缺失突变(984delC),导致317位密码子后移码突变,于444位密码子提前出现终止密码(H317fsX444);其母亲为此突变WASP基因携带者。另l例WAS患儿WASP基因第ll外显子,第1388位核苷酸由G替换为T(1388G→T),为无义突变,使第452位密码子提前变为终止密码(E452X)。其母亲无此突变WASP基因。结论 鉴定出两种新型WASP基因突变,WASP基因序列分析对于不典型和散发WAS的诊断及WASP突变基因携带者的检出有重要作用。     

X连锁中性粒细胞减少症1例报告并文献复习

目的总结WAS基因功能获得性突变致X连锁中性粒细胞减少症（XLN）的临床及分子特征,提高对该病的认识。方法报告1例先天性中性粒细胞减少症患儿的临床病史资料、常规免疫功能、骨髓细胞形态学特点及二代基因测序结果。对检测到的WAS基因突变行Sanger测序技术验证,采用流式细胞术分析患儿及其父母WAS蛋白表达,采用qPCR分析其WAS基因mRNA水平。分别以“X-linked neutropenia”和“X连锁中性粒细胞减少症”检索主要中英文数据库,检索时间为建库至2018年9月20日,提取文献中主要临床表型和基因型。结果男,6岁5月。自3月龄因发热发现WBC和ANC减少,后6年中WBC和ANC一直低于正常,约每月发热1次,伴反复呼吸道感染,有脓毒血症、EB病毒感染史,无湿疹。血常规示PLT计数和体积正常,WBC数量减少［（2.15±0.5）×109·L-1）］,ANC减少［(0.261±0.086)×109·L-1 ］,NK细胞减少［23.42×106·L-1（1.63%）］。骨髓增生活跃低水平,粒系成熟障碍。二代测序显示WAS基因9号外显子c.881T>C（p.I294T）错义突变,其母为该位点携带者。qPCR显示患儿WAS基因mRNA表达水平降低（为正常人的0.29倍）。流式检测显示患儿WAS蛋白表达量为正常对照的74.5%。PubMed数据库中筛选出XLN病例相关文献23篇,中文数据库0篇,总计18例（2个家系、3例散发）,报道4个突变位点（L270P、S272P、I290T、I294T）。加上本文1例（19例）进行临床资料总结,ANC数量减少占84.2%,单核细胞数量减少占72.2%,CD4+/CD8+比值倒置占56.3%,临床上主要表现为反复细菌感染和发热。结论 WAS基因功能获得性突变可导致先天性中性粒细胞减少,临床表现为反复感染,症状不突出,具有WAS mRNA表达减少、WASP蛋白仅轻微降低的分子特征。     

Wiskott-Aldrich综合征发病机制研究进展

Wiskott-Aldrich综合征(WAS)是一种少见的x连锁隐性遗传性免疫缺陷病,以血小板减少和体积缩小、湿疹、免疫缺陷三联征及易患自身免疫性疾病和恶性肿瘤为临床特点,由WAS蛋白基因突变致表达异常所致.WAS蛋白是仅在造血细胞系表达的胞质蛋白,主要参与细胞内信号传导及细胞骨架重新组合.最近研究发现,WAS蛋白基因突变除导致WAS、X连锁血小板减少症外,还可产生间断X连锁血小板减少症和X连锁中性粒细胞减少症.     

造血干细胞移植治疗Wiskott-Aldrich综合征一例报告及文献复习

目的 Wiskott-Aldrich综合征(WAS)是一种原发性免疫缺陷性疾病,严重病例预后不良.采用人类白细胞抗原(HLA)全相合的同胞骨髓移植成功治疗1例,特此总结并进行文献复习.方法 采用流式细胞仪检测WAS蛋白(WASP)表达和基因分析确诊WAS.患儿姐姐为人类白细胞抗原全相合骨髓供者,所采集骨髓单个核细胞数为4.38×108/kg,CD34+细胞3.78×106/kg患儿体重.采用白消安/环磷酰胺全清髓的预处理方案,环孢菌素单用预防移植物抗宿主病.移植后检测WASP表达和短串联重复序列(STR)作为植入证据.结果 患儿诊断:WAS,WASP(-IVS9+2T>C,WASP阴性).白消安/环磷酰胺预处理后骨髓回输;移植13 d中性粒细胞(ANC)绝对值0.8×109/L,移植15 d起血小板>50×109/L,1个月后正常.移植50 d起患儿WASP表达阳性,STR显示为供者DNA完全嵌合;随访至移植后510 d,患儿健康,WASP稳定表达.结论 结合病例和文献复习,人类白细胞抗原相合同胞骨髓移植治疗典型WAS近期预后较好.     

IVS6+5G>A found in Wiskott-Aldrich syndrome and X-linked thrombocytopenia in a Korean family

Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT) are caused by a mutation in the WAS gene on Xp11.22. We report two patients with IVS6+5G>A of WAS in a Korean family. The proband presented with classic WAS, whereas his maternal cousin had symptoms limited to XLT. Their mothers were proved to be carriers. The IVS6+5G>A mutation was reported to result in incomplete splicing of the donor site and typically associated with mild form of disease, XLT. Our observation of the intrafamilial variability of clinical manifestations of WAS further expands the genotype-phenotype correlations and suggests the presence of modifying genetic factors.     

Wiskott-Aldrich综合征6例临床分析

目的探讨Wiskott-Aldrich综合征(WAS)患儿的临床特点及治疗策略。方法对2000—2006于天津市儿童医院就诊的6例WAS患儿的临床表现进行临床表型评分,对患儿进行血常规、免疫球蛋白水平和骨髓常规检查,应用流式细胞仪对部分患儿T、B淋巴细胞比例进行检测。结果6例患儿均为典型WAS,除1例评3分外,其余5例均评4分。6例患儿家族史均为阴性。血常规检查显示平均血小板容积(MPV)减小,免疫球蛋白水平大致正常,但流式细胞检测显示,CD4 /CD8 升高或降低。肾上腺糖皮质激素治疗效果欠佳,应用大剂量静脉丙种球蛋白冲击可有短暂疗效,部分患儿需要输注血小板进行替代治疗。结论对出生后不久即有反复罹患血小板减少、湿疹和感染的男婴,应高度注意WAS的可能。对此类患儿应仔细询问家族史,注意MPV值的检测,积极控制感染,应用大剂量静脉丙种球蛋白进行冲击治疗,为患儿将来进行干细胞移植治疗创造机会。     

Analysis of clinical and molecular characteristics of Wiskott–Aldrich syndrome in 24 patients from 23 unrelated Chinese families

Zhang Z‐Y, Xiao H‐Q, Jiang L‐P, Zhou Y, Zhao Q, Yu J, Liu W, Yang X‐Q, Zhao X‐D. Analysis of clinical and molecular characteristics of Wiskott–Aldrich syndrome in 24 patients from 23 unrelated Chinese families.
Pediatr Allergy Immunol 2010: 21: 522–532.
© 2010 John Wiley & Sons A/S The clinical data of 24 children with Wiskott–Aldrich syndrome (WAS) from 23 unrelated Chinese families were reviewed in the present study. WAS protein (WASP) expression in peripheral blood mononuclear cells was examined by flow cytometry (FCM); WASP gene was amplified by PCR and directly sequenced to analyze mutations in the WASP gene in patients and their female relatives. FCM analysis of 21 patients showed that 18 cases were WASP‐negative, and three had partially WASP expression. WASP gene analysis revealed mutations in 23 patients, including five missense mutations, four nonsense mutations, four deletion mutations, three insertion mutations, six splice site mutations, and one complex mutation, among which, 20 unique mutations were detected, including seven novel mutations (168 C>A, 747–748del T, 793–797del C, 1185 ins C, Dup 1251–1267, 1277 insA and 1266 C>G; 1267–1269del C). Five WAS children underwent stem cell transplantation. After 2 months of transplantation, WASP expression was restored to normal in all five patients whereas one patient died of cytomegalovirus‐induced interstitial lung disease. WASP gene analysis can make a definite diagnosis of WAS and identify mutation carriers, beneficial for timely treatment and genetic counseling for children with WAS.     

WASP–WIP complex in the molecular pathogenesis of Wiskott–Aldrich syndrome

Wiskott–Aldrich syndrome (WAS) is an X‐linked primary immunodeficiency disease characterized by recurrent infection, thrombocytopenia, and eczema. The gene responsible for X‐linked WAS encodes the Wiskott–Aldrich syndrome protein (WASP), which is expressed in hematopoietic cells and which regulates T‐cell activation and cytoskeletal reorganization in T‐cell receptor (TCR) signaling. Here, I review my recent research on WASP and the WASP‐interacting protein (WIP) complex in T cells. I and my colleagues first established a diagnostic screening method using flow cytometry and genetic analysis, and elucidated the molecular pathogenesis in WAS patients with unique clinical manifestations. We investigated the mechanisms by which WASP is recruited to lipid rafts following TCR stimulation and to immunological synapses between antigen‐presenting cells and T cells. Subsequently, we elucidated the molecular mechanisms by which WASP is degraded by calpain and ubiquitinated by Cbl‐family proteins, which terminate WASP activation. More importantly, we found that WIP plays a critical role in WASP stability in T cells. These results provide new insights into the molecular pathogenesis of X‐linked WAS and have facilitated the identification of WIP deficiency as an autosomal recessive form of WAS.     

A unique presentation of Wiskott-Aldrich syndrome in relation to platelet size

Wiskott-Aldrich Syndrome (WAS) is a triad of immunodeficiency, eczema, and thrombocytopenia. Despite the heterogeneity of genetic and clinical findings, a correlation with small platelet size is routinely observed. Herein we describe a case with a unique phenotype that links normal mean platelet volume with the classic characteristics of this disease. The diagnosis was verified by genetic analysis showing a novel and de novo mutation. Our case illustrates that a high index of suspicion of WAS is warranted even in the setting of normal sized platelets.     

A case of Wiskott-Aldrich syndrome with de novo mutation at exon 4

Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by thrombocytopenia, eczema and immunodeficiency. Clinical features of the disease are highly varied; therefore, the diagnosis is sometimes difficult, especially in solitary cases or cases with milder forms of the disease. However, the identification of the WASP gene has made possible a definite WAS diagnosis for these cases. In this report, we present a 26-month-old boy who had received several ineffective treatments for chronic immune thrombocytopenic purpura. He was then suspected to have WAS because of the early onset of thrombocytopenia and small platelets. The diagnosis became definite with the detection of a de novo mutation at exon 4 of the WASP gene, Arg138Pro, through mutation analysis.     

Confirming or excluding the diagnosis of Wiskott-Aldrich syndrome in children with thrombocytopenia of an unknown etiology

Early diagnosis is an important factor in a better prognosis in patients with Wiskott-Aldrich syndrome (WAS), but it is not always easy to distinguish between WAS and immune thrombocytopenic purpura on clinical grounds. To confirm or to exclude a WAS diagnosis promptly for children with thrombocytopenia, the authors performed flow cytometric screening of Wiskott-Aldrich syndrome protein (WASP) for 10 children with thrombocytopenia of an unknown etiology. Five children were diagnosed with WAS, and the remaining 5 were diagnosed as having non-WAS causes of thrombocytopenia. There were no ambiguous results, and these were confirmed by genetic analysis. The authors conclude that screening by flow cytometry for WASP is recommended for boys with persistent thrombocytopenia of an unknown etiology.