Clinical features of Wiskott-Aldrich syndrome: an analysis of 13 cases / 中国当代儿科杂志

OBJECTIVE@#To study the clinical features of Wiskott-Aldrich syndrome (WAS) in children.@*METHODS@#A retrospective analysis was performed for the clinical data of 13 children with WAS.@*RESULTS@#All 13 children were boys, with a median age of onset of 3 months (range 1-48 months) and a median age of 24 months (range 1-60 months) at the time of diagnosis. Of the 13 children, only 3 had typical WAS and the remaining 10 children had X-linked thrombocytopenia (XLT). The mean WAS score was 2 (range 1-3), the mean platelet count was 20.5×10/L [range (13-46)×10/L], and the mean platelet volume was 8.1 fl (range 6.7-12.1 fl). Lymphocyte subsets and immunoglobulins were measured for 4 children, among whom 1 (25%) had a reduction in both the percentage of CD3T cells per lymphocyte and lymphocyte per nuclear cells, 1(25%) had a reduction in CD3CD56 NK cells. Among these 4 children, 1 (25%) had an increase in IgG, 2 (50%) had a reduction in IgM, 1 (25%) had a reduction in IgA, and 4 (100%) had an increase in IgE. A total of 14 gene mutations belonging to 13 types were found in 13 children, among which there were 9 missense mutations (65%), 2 splicing mutations (14%), 2 nonsense mutation (14%), and 1 frameshift mutation (7%). The median follow-up time was 39 months (range 3-62 months), and all 13 children survived.@*CONCLUSIONS@#Children with WAS often have a young age of onset, and most of them are boys. Major clinical features include thrombocytopenia with a reduction in platelet volume. Missense mutation is the main type of gene mutation.

Analysis of Correlation of WAS Gene Mutations with Clinical Phenotype / 中国实验血液学杂志

OBJECTIVE@#To investigate the gene mutation of patients with WAS gene defect and its correlation with clinical manifestations.@*METHODS@#Thirty-one patients consulted in Children's Hospital of Soochow University from January 2013 to February 2018 were enrolled in this study. The hot pot mutations of WAS gene in 31 patients were detected and related clinical phenotypes were analyzed retrospectively.@*RESULTS@#All patients were male. The median onset age was 1 month (range, 0-83 months). Nine mutants were reported as novel mutations among 25 mutants detected in 31 patients, including c.1234_1235dupCC, c.1093-1097delG, c.28-30dupC, c.436G>T, c.273 + 10_273 + 11dupCC, c.995_996insG, c.1010T>A, c.332_333delCC and c.683C>T mutations. There were 25 cases of classic WAS which mutations included missense mutation, deletion mutation, insertion mutation, splicing mutation and nonsense mutation, 2 cases of X-linked thrombocytopenia (XLT) were induced by missense mutation, 1 case of intermittent X-linked thrombocytopenia (IXLT) was induced by splicing mutation, 2 cases of X-linked pancytopenia were induced by missense mutation. Intravenous immunoglobulin (IVIG) and glucocorticoid therapy in IXLT patient was effective, and remission could be sustained, platelets could be increased in the short-term in treated XLT patients, but only a small part of classic WAS patients(8.0%) showed transient response to it, the IVIG and glucocorticoid therapy did not improve the status of platelet in XLP patients. Immune laboratory examination showed that CD3 was decreased in 60.0% patients, CD19 was decreased in 12.0% patients, and CD56CD16 in 4 patients was decreased, accounting for 16.0%. Out of 24 patients, 22 patients were alive after treated with hematopoietic stem cell transplantation (HSCT), 4 patients who were not given HSCT died of brain bleeding and severe infection, 1 patient diagnosed as IXLT got remission and survived.@*CONCLUSION@#WAS gene defect is an important basis for the diagnosis of WAS and related diseases. IVIG plus glucocorticoid therapy is less effective for fewer patients, the HSCT is an effective treatment for WAS.

Analysis of WAS gene mutation in a Chinese family affected with Wiskott-Aldrich syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutation of the WAS gene in a Chinese family affected with Wiskott-Aldrich syndrome.</p><p><b>METHODS</b>Peripheral blood samples were collected from the proband and his family members. All exons and flanking regions of the WAS gene were subjected to PCR amplification - Sanger sequencing as well as restriction endonuclease analysis. Plasma level of B-cell activating factor (BAFF) was also determined for all family members.</p><p><b>RESULTS</b>A hemizygous mutation (c.257G>A) of the WAS gene was identified in all patients from the family, for which the patient's mother was heterozygous. The same mutation was not found among healthy members of the family. Compared with unaffected members, all patients had a higher level of BAFF.</p><p><b>CONCLUSION</b>The c.257G>A mutation of the WAS gene probably underlies the Wiskott-Aldrich syndrome in this family.</p>

Síndrome de Wiskott-Aldrich com plaquetas de volume normal / Wiskott-Aldrich syndrome with normal-sized platelets

A Síndrome de Wiskott-Aldrich (WAS) é uma imunodeficiência congênita ligada ao cromossomo X, caracterizada por mutações no gene WAS, responsável pela proteína WASP. As principais manifestações clínicas são trombocitopenia com plaquetas de volume reduzido, eczema, infecções recorrentes e maior incidência de doenças autoimunes e neoplasias. Relatamos o caso de um paciente do sexo masculino com sintomas clássicos desta síndrome (eczema, trombocitopenia e infecções recorrentes), porém com plaquetas de volume normal. Existem poucos relatos desta síndrome em pacientes com plaquetas de volume normal, o que atrasou o encaminhamento do paciente ao imunologista, o qual foi tratado como portador de Síndrome de Evans e dermatite atópica até os quatro anos de idade. A confirmação diagnóstica foi por teste genético. O diagnóstico precoce possibilita profilaxia com antibioticoterapia e uso de imunoglobulina endovenosa, devido ao risco de infecções graves, e encaminhamento para transplante de células-tronco hematopoiéticas, que até o momento é o único tratamento curativo. A suspeita clínica deve existir em pacientes com trombocitopenia inexplicável, mesmo se as plaquetas tiverem o tamanho normal, associada às outras manifestações da doença.

Wiskott-Aldrich syndrome (WAS) is an X-linked congenital immunodeficiency characterized by mutations in the WAS gene of the WASP protein. The main clinical manifestations are thrombocytopenia with small-sized platelets, eczema, recurrent infections and a higher incidence of autoimmune diseases and cancer. We report the case of a male patient with classical symptoms of this syndrome (eczema, thrombocytopenia and recurrent infections), however presenting platelets with normal size. There are few reports of this syndrome in patients with normal-sized platelets, which delayed our patient's referral to the immunologist. The patient received treatment for Evans syndrome and atopic dermatitis until he was four years old. Confirmation of WAS diagnosis was made by genetic testing. Early diagnosis allows prophylactic treatment with antibiotics and the use of intravenous immunoglobulin due to the risk of serious infections, in addition to referral for hematopoietic stem cell transplantation, which is the only curative treatment available so far. WAS should be suspected when patients develop unexplained thrombocytopenia even with normal-sized platelets, especially in the presence of other manifestations.

Use of positive pressure in preoperative and intraoperative of bariatric surgery and its effect on the time of extubation / Utilização da pressão positiva no pré e no intraoperatório de cirurgia bariátrica e seus efeitos sobre o tempo de extubação / Utilización de la presión positiva en el pre- y en el intraoperatorio de cirugía bariátrica y sus efectos sobre el tiempo de extubación

BACKGROUND AND OBJECTIVE: To investigate the influence of intraoperative and preoperative positive pressure in the time of extubation in patients undergoing bariatric surgery. METHOD: Randomized clinical trial, in which 40 individuals with a body mass index between 40 and 55 kg/m2, age between 25 and 55 years, nonsmokers, underwent bariatric surgery type Roux-en-Y gastric bypass by laparotomy and with normal preoperative pulmonary function were randomized into the following groups: G-pre (n = 10): individuals who received treatment with noninvasive positive pressure before surgery for 1 h; G-intra (n = 10): individuals who received positive end-expiratory pressure of 10 cm H2O throughout the surgical procedure; and G-control (n = 20): not received any preoperative or intraoperative intervention. Following were recorded: time between induction of anesthesia and extubation, between the end of anesthesia and extubation, duration of mechanical ventilation, and time between extubation and discharge from the post-anesthetic recovery. RESULTS: There was no statistical difference between groups. However, when applied to the Cohen coefficient, the use of positive end-expiratory pressure of 10 cm H2O during surgery showed a large effect on the time between the end of anesthesia and extubation. About this same time, the treatment performed preoperatively showed moderate effect. CONCLUSION: The use of positive end-expiratory pressure of 10 cm H2O in the intraoperative and positive pressure preoperatively, influenced the time of extubation of patients undergoing bariatric surgery. .

JUSTIFICATIVA E OBJETIVO: investigar a influência do uso da pressão positiva nas vias aéreas intraoperatória e pré-operatória no tempo de extubação de pacientes submetidos à cirurgia bariátrica. MÉTODO: Trata-se de ensaio clínico randomizado, no qual 40 indivíduos com índice de massa corporal entre 40 e 55 kg/m2, idade entre 25 e 55 anos, não tabagistas, submetidos à cirurgia bariátrica do tipo derivação gástrica em Y de Roux por laparotomia e com prova de função pulmonar pré-operatória dentro da normalidade foram randomizados nos seguintes grupos: G-pré (n = 10): indivíduos que receberam tratamento com pressão positiva não invasiva antes da cirurgia, durante uma hora, G-intra (n = 10): indivíduos que receberam Positive End-expiratory Pressure de 10 cm H2O durante todo o procedimento cirúrgico e G-controle (n = 20): não receberam qualquer tipo de intervenção pré ou intraoperatória. foram anotados os seguintes tempos: tempo decorrido entre a indução anestésica e a extubação, entre o término da anestesia e extubação, tempo de ventilação mecânica, e tempo entre a extubação e a alta da Recuperação Pós-Anestésica. RESULTADOS: Não houve diferença estatística entre os grupos, porém quando aplicado ao Coeficiente de Cohen, o uso da Positive End-expiratory Pressure de 10 cm H2O no intraoperatório mostrou um efeito grande sobre o tempo entre o término da anestesia e a extubação. Sobre este mesmo tempo, o tratamento realizado no pré-operatório apresentou efeito moderado. CONCLUSÃO: O uso da Positive End-expiratory Pressure de 10 cm H2O no intraoperatório e da pressão positiva no pré-operatório, pode influenciar o tempo de extubação de pacientes submetidos à cirurgia bariátrica. .

JUSTIFICACIÓN Y OBJETIVO: Investigar la influencia del uso de la presión positiva en las vías aéreas intraoperatoria y preoperatoria en el tiempo de extubación de pacientes sometidos a la cirugía bariátrica. MÉTODO: Se trata de un ensayo clínico aleatorizado, en el cual 40 individuos con IMC entre 40 y 55 kg/m2, edad entre 25 y 55 años, no fumadores, sometidos a cirugía bariátrica del tipo derivación gástrica en Y de Roux por laparotomía y con prueba de función pulmonar preoperatoria dentro de la normalidad fueron aleatorizados en los siguientes grupos: G-pre (n = 10): individuos que recibieron tratamiento con presión positiva no invasiva antes de la cirugía durante una hora; G-intra (n = 10): individuos que recibieron PEEP de 10 cm H2O durante todo el procedimiento quirúrgico y G-control (n = 20): no recibieron ningún tipo de intervención pre- o intraoperatoria. Fueron anotados los siguientes tiempos: tiempo trascurrido entre la inducción anestésica y la extubación, entre el fin de la anestesia y la extubación, tiempo de ventilación mecánica, y tiempo entre la extubación y el alta de la sala de recuperación postanestésica. RESULTADOS: No hubo diferencia estadística entre los grupos, sin embargo cuando se aplicó el coeficiente de Cohen, el uso de la PEEP de 10 cm H2O en el intraoperatorio mostró un efecto importante sobre el tiempo entre el término de la anestesia y la extubación. Sobre ese mismo tiempo, el tratamiento realizado en el preoperatorio presentó un efecto moderado. CONCLUSIÓN: El uso de la PEEP de 10 cm H2O en el intraoperatorio y de la presión positiva en el preoperatorio puede influir en el tiempo de extubación de pacientes sometidos a cirugía bariátrica. .

The gene mutation analysis of a Wiskott-Aldrich syndrome family with normal mean platelet volume / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the history of a Wiskott- Aldrich syndrome (WAS) family with normal mean platelet volume (MPV), analyse the WASP gene mutation of to better understand its clinical characteristics.</p><p><b>METHODS</b>A four- generation WAS family histories of 22 members were investigated. Peripheral blood samples were collected from propositus and his mother to analyse all exon mutations of WASP gene using sanger sequencing.</p><p><b>RESULTS</b>The MPV of both propositus and his elder brother were normal. The patient's clinical score was 5, antibodies to PM-Scl, PCNA and PO were positive with very high level of ASO, the patient co- suffered from autoimmune disease, anemia, abnormal renal function, fungal infection and scleritis. A homozygous mutation (C>T) was found at 173 bp of exon 2, corresponding to amino acids Pro (P) 58 abnormally changed to Leu (L). His mother was the carrier of the mutation. Of 112 blood diseases- related genes, mutation frequencies of CBL, CREBBP, DNM2 and ADAMTS13 were higher than normals.</p><p><b>CONCLUSION</b>This was the first report the phenotype 173C>T mutation of WASP without eczema, but with normal MPV and autoimmune disease in Chinese, WAS should be recognized earlier and diagnosed correctly by genomic methods.</p>

Clinical features and genotype analysis of 132 patients with Wiskott-Aldrich syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical and immunological laboratory features, gene mutations, treatment and prognosis in children with Wiskott-Aldrich syndrome (WAS).</p><p><b>METHOD</b>The clinical, laboratory characteristics, treatment and prognosis of 132 children with WAS, who visited Children's Hospital of Chongqing Medical University from April 2000 to June 2015, were analyzed retrospectively.</p><p><b>RESULT</b>All patients were male. The median age of disease onset was 15 days and the median age at diagnosis was 10 months. Of the 132 cases, 112 had classic WAS, 20 had X-linked thrombocytopenia (XLT). The median platelet count was 23×10(9)/L. All cases had the clinical characteristics of WAS including bleeding, eczema, and being susceptible to infection. The initial symptoms include hemorrhage (75.0%) and eczema (16.7%). Twenty-one cases had autoimmune diseases and one patient had leukemia. WAS protein (WASP) expression in 115 cases were measured by flow cytometry, 88 cases were negative, in 12 cases WASP decreased, in 5 cases it was normal, 10 cases had bimodal distribution. Eighty-one kinds of mutations were found in 122 families, including eight kinds of hot-spot mutations, which were 290 C> N / 291G> N (R86C / H / L), 665 C> T (R211X), 155 C> T (R41X), 168 C> T (T45 M), IVS1+ 1 g> t/ a, IVS6 + 5 g> a, IVS8 + 1 g> a and IVS8 + 1to + 6del gtga. Meantime, 29 kinds of novel mutations were found, which were 321T>C, 415C>A, 471C>T, 102-105delC, 521 del C, 1330 del A, IVS2-2 a>c, 168 C>A/1412 C> T, exon1-2 del/1412 C>T, and so on. The proportion of CD3(+) T cells (31.3%), helper T cells (37.3%) and cytotoxic T cells (38.6%) in the peripheral blood declined. The serum levels of IgG (51.1%), IgA (43.3%) and IgE (40.0%) increased, IgM (25.6%) decreased. Of the 132 cases, 72 remain survived, of whom 36 cases received hematopoietic stem cell transplantation (HSCT), 14 patients with classic WAS received intravenous immunoglobulin (IVIG) therapy. With regular IVIG therapy, the frequency of infections was reduced and the patients' symptoms were improved.</p><p><b>CONCLUSION</b>The clinical characteristics of Wiskott-Aldrich syndrome were early age of onset, microthrombocytopenia, eczema and recurrent infections. The proportion of T lymphocyte declined, the serum levels of IgG, IgA, and IgE increased, and level of IgM decreased in a part of patients. The detection of WAS gene mutation and WAS protein detection was the key diagnostic methods. Regular IVIG can gain more time for children who will receive HSCT and improve their quality of life.</p>

Mutation analysis of WASP gene and prenatal diagnosis of Wiskott-Aldrich syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and an increased incidence of autoimmunity and malignancies. The patients always have a severe clinical phenotype that can result in death if not diagnosed and treated early in life. The treatment of choice with the best outcome is hematopoietic stem cell transplantation, preferably from a matched related donor. But uncertain treatment effect and high treatment cost limit its clinical application. It is the best strategy that avoiding birth of a fetus with defect through prenatal diagnosis at present. This study aimed to analyze the mutation of WASP gene in 4 Chinese families with WAS and to provide prenatal diagnosis for the high-risk fetus.</p><p><b>METHOD</b>The probands of the four WAS families were all males, one of whom was deceased but had a family history and clinical datas integrated. All the patients were detected with blood routine tests, immunological tests and bone marrow examination. PCR and bilateral direct sequencing of PCR product was carried out in the regions of exon and exon-intron boundaries of WASP gene for 3 probands, 4 mothers and 100 unrelated healthy individuals as control. Prenatal diagnosis was provided for the two fetuses at the first trimester by mutation analysis.</p><p><b>RESULT</b>Four WASP gene mutations were detected: c.91A > G (p.E31K), c.665C > T (p.R211X), c.397G > A (p.E133K), c.952-953delCC (p. P317fsX18), among which c.952-953delCC (p. P317fsX18) was first reported. Mothers in Family 2, 3 and 4 were carriers of WASP gene mutation, but family 1 was considered as a de-novo mutation. None of the 100 unaffected subjects had the above mutants. Prenatal diagnosis indicated that the fetus in family 2 was male and carried the same mutation as the proband, so the fetus was presumably to be a patient. The parents decided to receive an induced abortion. Following the termination of the pregnancy, the result of gene analysis of the aborted tissues was consistent with prenatal diagnosis. The fetus in family 3 was normal male confirmed by normal test results six months after birth.</p><p><b>CONCLUSION</b>The 4 mutations of the WASP gene probably were causative to the families of WAS, among which c.952-953delCC was reported for the first time. Prenatal diagnosis by DNA sequencing is the effective method to avoid birth of WAS patient.</p>

Estudo de células T regulatórias em camundongos deficientes em WASP / T Cell study regulatory in mice disabled in wasp

A Síndrome de Wiskott-Aldrich (WAS) é uma imunodeficiência associada a infecçõesrecorrentes, câncer e autoimunidade. Ela é causada por mutações no gene que codifica amolécula WASP (“Wiskott-Aldrich syndrome protein”), envolvida na reorganização docitoesqueleto de actina em células de origem hematopoiética. Camundongos deficientes emWasp (WKO) apresentam migração leucocitária e proliferação linfocitária deficientes, edesenvolvem colite. Esse processo inflamatório está associado a eventos deimunodesregulação importantes, como participação exacerbada de células Th2 e diminuiçãode células T regulatórias (Treg) no timo e baço desses animais. Assim, esse trabalho visacaracterizar as células Treg tímicas e avaliar os mecanismos potencialmente relacionados àdiminuição dessas células no timo de camundongos WKO. Nas análises de CD25 e CD122,importantes para a diferenciação de células Treg, observamos uma diminuição na expressãode CD25 na população de células CD4+CD8-Foxp3+, além de uma diminuição no númeropercentual dessa população. As células Treg tímicas de animais WKO apresentaram níveisnormais de CD122 e de moléculas de ativação, mas mostraram uma diminuição na expressãoda molécula funcional CD39, responsável pela hidrólise tecidual de ATP. Saliente-se aindaque estudos sobre a morte celular ex-vivo, a transmigração in vitro mediada por S1P e alocalização intratímica de células Treg não revelaram diferenças significativas na comparaçãoentre animais WKO e seus controles. De forma interessante, precursores tímicos de célulasTreg CD4+CD8-CD25+Foxp3-também se apresentaram diminuídos em WKO, assim como ascélulas Treg Helios...

The Wiskott-Aldrich syndrome (WAS) is an immunodeficiency associated with recurrentinfections, malignancies and autoimmunity. It is caused by mutations in the gene encodingWASP (Wiskott-Aldrich syndrome protein), a molecule involved in the actin cytoskeletonrearrangements in hematopoietic cells. Wasp deficient mice (WKO) show impaired leukocytemigration and lymphocyte proliferation, and also develop colitis. This inflammatory processis associated with important immunodysregulation events, such as exacerbated participationof Th2 cells and decreased numbers of regulatory T cells (Treg) in the thymus and spleen ofthese animals. Thus, this study aims to characterize the thymic Treg cells and evaluate thepotential mechanisms involved in the decreased number of these cells in the thymus of WKOmice. In the analysis of CD25 and CD122, important molecules for the differentiation of Tregcells, we observed a decreased expression of CD25 in the CD4+CD8-Foxp3+ cells, and adecrease in the percentage number of this population. WKO Treg cells showed normal levelsof CD122 and activation molecules, but present a decreased expression of the functionalmarker CD39, molecule responsible for ATP hydrolysis in the tissues. Also, it is important topoint out that studies about ex vivo cell death, S1P-mediated in vitro transmigration andintratymic localization of Treg cells revealed no significant differences between WKOanimals and their controls. Interestingly, both CD4+CD8-CD25+Foxp3- Treg precursors andHelios+ Treg cells are reduced in WKO thymus. More over, we also observed an importantincrease in the basal expression of pSTAT-5 in Treg cells of WKO mice and their precursors.Altogether, our results indicate an important role of Wasp in thymic Treg cell development,and point to the necessity of further studies on the mechanisms involved in the generation ofTreg cells in WKO mice...

Analysis of prenatal diagnosis for seven high-risk fetuses with Wiskott-Aldrich syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the value of gene analysis of amniotic fluid exfoliated cells and WASP detection from cord blood in prenatal diagnosis of high-risk fetus with Wiskott-Aldrich syndrome.</p><p><b>METHOD</b>Seven patients with Wiskott-Aldrich syndrome were diagnosed by gene analysis and WASP detected by flow cytometry from 2008 to 2010. After detailed inquiry for medical history and gene analysis of related family members, seven pedigree trees were drawn, including 15 carriers of abnormal genes. From 2008 to 2011, seven samples of amniotic cell gotten by amniocentesis were collected from seven high-risk pregnant women with abnormal gene during 18 to 20 gestational weeks. WASP gene was amplified by polymerase chain reaction (PCR) from DNA of amniotic cell gotten and sequencing was performed directly on the PCR products forward and reversely. Embryo blood sample was collected from one high-risk fetus by needle puncture of umbilical blood vessel and WASP expression was detected by flow cytometry. Karyotyping was performed in amniotic cell gotten cultivated by orthotopic slice and G band staining. Gene analysis of WASP, WASP expression detected by flow cytometry and evaluation of immune function were reexamined in high-risk fetus after delivery.</p><p><b>RESULT</b>Amniocentesis and culture of amniotic cell succeeded in all the seven fetuses. Gene analysis and karyotyping showed that one male fetus and four female fetuses were normal and two female fetuses were carriers. WASP expression detected from embryo blood sample of the patient was normal. After delivery, the result of gene analysis, WASP detection and evaluation of immune function was the same as that of prenatal diagnosis.</p><p><b>CONCLUSION</b>Karyotyping, gene analysis and WASP detection of cord blood can provide reliable service of prenatal diagnosis for high-risk pregnant women with Wiskott-Aldrich syndrome.</p>

Síndrome de Wiskott-Aldrich / Wiskott - Aldrich syndrome

Objetivo: Descrever um caso de síndrome de Wiskott-Aldrich, enfatizandoa importância do diagnóstico precoce de uma imunodeficiênciarara, para seu tratamento adequado.Descrição do caso: Criança do sexo masculino, que aos seismeses de idade apresentou eczema em face e pescoço. Três mesesapós, evoluiu com piora, sendo internado com infecção secundária doeczema. Aos dez meses foi novamente internado por otite média comsecreção sanguinolenta, sangramento oral e lesões em pele com sufusõeshemorrágicas. Com um ano foi internado pela terceira vez, devidoà diarreia sanguinolenta, evoluindo com sepse. Exames laboratoriaisevidenciaram plaquetopenia e anemia, além de número reduzido delinfócitos T CD4+ e CD8+. A pesquisa para proteína da síndrome deWiskott-Aldrich foi ausente.Discussão: A Síndrome de Wiskott-Aldrich (WAS) é uma imunodeficiênciarara, ligada ao X, com manifestações clínicas característicasque incluem trombocitopenia com plaquetas pequenas, eczema, infecçõesrecorrentes e incidência aumentada de manifestações autoimunese malignidades. O diagnóstico precoce é muito importante para umtratamento adequado. Até o momento, a única terapia curativa é otransplante de células tronco.

Objective: Describe a case of Wiskott-Aldrich syndrome, emphasizingthe importance of early diagnosis of a rare immunodeficiency, for itsappropriate treatment.Case description: Male child, who at six months of age presentedeczema in face and neck. Three months later, progressed to worse,being admitted with secondary infection of the eczema. At ten monthswas again hospitalized due to otitis media with drainage of blood, oralbleeding and skin lesions with hemorrhagic suffusions. With one yearold was hospitalized for the third time, due to bloody diarrhea, evolvingto sepsis. Laboratory tests showed anemia and thrombocytopenia, andreduced number of CD4 and CD8 T lymphocytes. The search for theWiskott-Aldrich syndrome protein was absent.Discussion: The Wiskott-Aldrich Syndrome (WAS) is a rareimmunodeficiency, X-linked, with clinical features that includethrombocytopenia with small platelets, eczema, recurrent infections andincreased incidence of autoimmune manifestations and malignancies.Early diagnosis is very important for appropriate treatment. So far, theonly curative therapy is the transplantation of stem cells.

Analysis of clinical features and gene mutations in 6 patients with Wiskott-Aldrich syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate clinical features, laboratory alterations and gene mutations of 6 patients with Wiskott-Aldrich syndrome (WAS).</p><p><b>METHODS</b>T lymphocyte subtypes were measured by flow cytometer. The routine blood tests including platelet count and mean platelet volume were performed by complete blood analyzer Sysmex XE2100. Serum immunoglobulin was measured by immunoturbidimetry. Mutations in WAS protein (WASP) gene (including all the exons and exon-intron boundaries and 3', 5' untranslation region) of 6 patients and their family members were identified by PCR and sequencing.</p><p><b>RESULTS</b>The patients presented with petechiae, easy bruise, eczema, bloody diarrhea, recurrent infection and fever, and the clinical scores were 3 or 4. They were thrombocytopenia with smaller mean platelet volume, anemia and leukocytosis. Megakaryocyte number was normal or slightly increased in bone marrow. In the probands, the percentage of CD3+ T cells was decreased, the CD4+/CD8+ ratio was abnormal, while the fractions of CD19+ and CD16+ CD56+ cells were in normal range. In most of the patients, the serum levels of IgG and IgA were increased. Six mutations were identified in the patients, including 10250 C-->T, and five novel mutations: 6783 C-->G,10216-10221 Ins G, 9964 Del T,10192-10203 Del GCCTGCCGGGG and 10052-10059 del GCTACTG. The 6783 C-->G in exon 3 resulted in premature stop at Tyr102, and the remaining four mutations in exon 10 resulted in frame shift and premature stop.</p><p><b>CONCLUSION</b>The main characteristics of these WAS patients were thrombocytopenia with smaller mean platelet volume and immunological disturbance. Their gene mutations were deletion, insertion or nonsense mutations. All the patients had been misdiagnosed as ITP, indicating the importance of differential diagnosis.</p>

WASP gene mutation analysis of a family of X-linked thrombocytopenia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study investigated the history and gene mutations of a family with X-linked thrombocytopenia, in order to understand the clinical characteristic and molecular pathogenesis of the disease.</p><p><b>METHODS</b>A three-generation X-linked thrombocytopenia family with 13 family members was investigated using PCR-DNA direct sequencing method to screen the exons of WASP gene for mutation analysis.</p><p><b>RESULTS</b>The WASP gene sequencing of the proband revealed a missense mutation in exon 2 (G291A), resulting in a change of amino acid 86 from arginine to histidine. The patient's mother was the carrier of the heterozygosis mutation in X-chromosome.</p><p><b>CONCLUSIONS</b>WASP mutations may be attributed to the molecular mechanism of X-linked thrombocytopenia. G291A is one of the mutations of WASP.</p>

Phenotypic and Genotypic Correction of WASP Gene Mutation in Wiskott-Aldrich Syndrome by Unrelated Cord Blood Stem Cell Transplantation

We present two cases of Wiskott-Aldrich syndrome (WAS), in which nonsense mutations in the WASP gene were corrected phenotypically as well as genotypically by unrelated cord blood stem cell transplantation (CBSCT). Two male patients were diagnosed with WAS at the age of 5-month and 3-month and each received unrelated CBSCT at 16-month and 20-month of age, respectively. The infused cord blood (CB) units had 4/6 and 5/6 HLA matches and the infusion doses of total nucleated cells (TNC) and CD34+ cells were 6.24x10(7)/kg and 5.08x10(7)/kg for TNC and 1.33x10(5)/kg and 4.8x10(5)/kg for CD34+ cells, for UPN1 and UPN2, respectively. Complete donor cell chimerism was documented by variable number tandem repeat (VNTR) with neutrophil engraftment on days 31 and 13 and platelets on days 58 and 50, respectively. Immunologic reconstitution demonstrated that CBSCT resulted in consistent and stable T-, B-, and NK-cell development. Flow cytometric analysis for immunologic markers and sequence analysis of the WASP gene mutation revealed a normal pattern after CBSCT. These cases demonstrate that CBs can be an important source of stem cells for the phenotypical and genotypical correction of genetic diseases such as WAS.

A Familial Case of Wiskott-Aldrich Syndrome with a Hotspot Mutation in Exon 2 of the WAS Gene

The Wiskott-Aldrich syndrome (WAS) is a severe X-linked disorder characterized classically by thrombocytopenia, immunodeficiency, and eczema. The phenotype observed in this syndrome is caused by mutation in the WAS gene. Peripheral blood DNAs were isolated from an 18-month-old boy with WAS and his mother, maternal uncle, and maternal grandmother. Genetic analysis for the detection of a mutation of WAS gene was performed by polymerase chain reaction-single strand conformational polymorphism analysis (PCR-SSCP) and direct sequencing of the PCR product. In PCR-SSCP, the patient and his maternal uncle had an abnormal shift band, which was not found in normal controls, and his mother and maternal grandmother showed heterozygous bands. In direct sequencing analysis, the patient with WAS had CGC-->CAC point mutation in exon 2 that resulted in an amino acid change in codon 86 (Arg86His). The present study identified a gene mutation responsible for WAS at a mutation hotspot of the WAS gene in a Korean family.

Diagnostic Approach of Wiskott-Aldrich Syndrome / 소아과

OBJECTIVE: A probable diagnosis of Wiskott-Aldrich syndrome(WAS) should be considered in any boy presenting with unusual bleeding, congenital thrombocytopenia and small platelets. The definitive diagnosis of WAS is usually made by the detection of WASP gene mutation or a decrease or absence of the WAS protein(WASP) in blood cells using molecular genetic analysis. However, these methods are too time-consuming and difficult. In this study, therefore, we tried to compare various diagnostic methods and establish the molecular screening steps for the definitive diagnosis of WAS. METHODS: Peripheral blood was drawn from WAS patient with clinical characteristic symptoms, and analyzed with automated complete blood cell count analysis, immunological analysis, and molecular analysis. The morphologic change of WAS patient's blood cell membrane was examined by scanning electron microscopy(SEM). Protein analysis for the expression of WASP protein in PBMC cells was evaluated by FACS and Western immunoblot. Genetic analysis for the detection of a mutation of the WASP gene was performed by polymerase chain reaction-single strand conformational polymorphism analysis(PCR-SSCP) and direct sequencing of PCR products. RESULTS: In addition to microthrombocytopenia, our investigation revealed morphologic defects in WAS lymphocytes by SEM and abnormal mobility shifting in WAS patients by PCR-SSCP. Sequencing the WASP gene detected a specific single base mutation in exon 2, resulting in missense substitution of adenine for guanine 208(G208A, Gly70Arg). FACS and Western immunoblot demonstrated absent expressions of WASP in WAS patients and reduced expression of WASP in carrier when compared with normal controls. CONCLUSION: From these results, we suggested the following diagnostic approaches in patients suspected of having WAS. Protein-based analysis such as FACS and Western immunoblot should be employed as the first line of investigation. The second line genetic analyses should employ second- step approaches with localization of mutation by screening exons, typically by PCR-SSCP, followed by direct sequencing.

Identification of two novel WASP gene mutations in 3 boys with Wiskott-Aldrich syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>The Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency caused by mutations in the WAS protein (WASP) gene. The disease is characterized by recurrent infections, eczema, and thrombocytopenia with small platelets, and it is known to be associated with extensive clinical variability, and mutation studies indicated that genotypes are also highly variant among WAS patients. The present study was conducted to identify the mutation types of Wiskott-Aldrich syndrome protein (WASP) gene in 3 boys suffering from Wiskott-Aldrich syndrome.</p><p><b>METHODS</b>Based on the typical clinical manifestations of Wiskott-Aldrich syndrome including thrombocytopenia, eczema, and recurrent infections and scanning electron micrographs, 3 patients were suspected of having WAS. The WASP gene of the 3 patients and their mothers were detected by PCR-direct sequencing analysis.</p><p><b>RESULTS</b>By sequence analysis using sense and antisense primer separately, the authors found two novel WASP gene mutations. For the twin brothers, a C deletion at nucleotide 984 was detected in exon 10 of WASP gene (984delC). The consequence of the C deletion involved frameshift mutation after H317 and premature stop at 444 (H317fsX444). Their mother was a carrier of the mutated WASP gene. For another WAS patient, a nonsense mutation with nucleotide substitution of G to T at position 1388 (1388G-->T) in exon 11 of WASP gene, led to premature translational termination at amino acid position 452 (E452X). His mother had not been found to have WASP gene mutation.</p><p><b>CONCLUSION</b>Genetic analysis is useful in definite diagnosis of Wiskott-Aldrich syndrome patients and in carrier detection and prenatal diagnosis, especially of atypical or sporadic WAS patients.</p>