Abscesos cerebrales por Dermacoccus nishinomiyaensis como primera manifestación de enfermedad granulomatosa crónica / Dermacoccus nishinomiyaensis brain abscesses as the first manifestation of chronic granulomatous disease

La enfermedad granulomatosa crónica es una inmunodeficiencia primaria poco frecuente, que secaracteriza por defectos en alguna de las subunidades del complejo enzimático nicotinamida adeninadinucleótido fosfato oxidasa, que ocasiona un déficit en la generación de anión superóxido por losfagocitos. Dentro de este grupo, la forma ligada al X es la más frecuente. Se reporta el caso de una paciente de sexo femenino de 2 años con enfermedad granulomatosa crónica autosómica recesiva, con mutación en gen CYBA, quien presentó manifestación inicial de la enfermedad con abscesos cerebrales ocasionados por un germen oportunista (Dermacoccus nishinomiyaensis). Esta infección permitió la sospecha diagnóstica temprana, por lo que recibió el tratamiento y la profilaxis en forma oportuna. Actualmente, se encuentra libre de infecciones, a la espera del trasplante de células progenitoras hematopoyéticas.

Chronic granulomatous disease is a rare primary immunodeficiency characterized by defects in one of the subunits of the nicotinamide adenine dinucleotide phosphate oxidase enzyme complex, which causes a deficiency in the capacity of phagocytes to generate superoxide anion. Within this group, the X-linked form is the most frequent. Here we report the case of a 2-year-old female patient with autosomal recessive chronic granulomatous disease, with a mutation in the CYBA gene, whose initial manifestation was brain abscesses caused by an opportunistic microorganism (Dermacoccus nishinomiyaensis). The infection led to an early diagnostic suspicion, so treatment and prophylaxis were administered in a timely manner. Currently, she is infectionfree, awaiting hematopoietic progenitor cell transplantation.

Enfermedad granulomatosa crónica: fisiopatología genética y molecular. Pruebas diagnósticas vigentes / Chronic granulomatous disease: genetic and molecular pathophysiology. Effective diagnostic tests

Introducción: La enfermedad granulomatosa crónica es una inmunodeficiencia primaria congénita del sistema inmune innato, originada por defectos en el complejo enzimático nicotinamida adenina dinucleótido fosfato oxidasa presente en células fagocíticas. Estos defectos funcionales causan incapacidad para producir especies reactivas del oxígeno en los fagocitos, que afectan la eliminación de algunos microorganismos patógenos dentro del fagolisosoma. El diagnóstico de esta enfermedad se realiza actualmente mediante la prueba de 1,2,3-dihidrorodamina asistida por citometría de flujo multiparamétrica, o la tinción de fagocitos con nitroazul de tetrazolio asistida por microscopio óptico. Objetivos: Describir los aspectos fisiopatológicos y moleculares de la enfermedad granulomatosa crónica; y discutir aspectos relacionados con las pruebas de diagnóstico antes mencionadas. Métodos: Se realizó una investigación bibliográfica-documental a partir de artículos científicos publicados desde 1933 hasta 2018, para ello fueron consultadas las bases de datos SciELO, PubMed y Springer. Desarrollo: Se exponen las características fisiopatológicas de la enfermedad granulomatosa crónica, así como la relación entre las mutaciones genéticas más abundantes en la población afectada y la gravedad de las manifestaciones clínicas que presentan los pacientes. Además, se analizan críticamente los beneficios y las deficiencias de dos técnicas que se utilizan actualmente para diagnosticar la enfermedad. Conclusiones: La enfermedad granulomatosa crónica puede generar consecuencias inmunológicas e inflamatorias graves, que se hallan en consonancia con las características genéticas expresadas en el complejo enzimático dañado. El diagnóstico de la enfermedad resulta más confiable, exhaustivo y específico, mediante la citometría de flujo y su prueba de 1,2,3-dihidrorodamina(AU)

Introduction: Chronic granulomatous disease is a congenital primary immunodeficiency of the innate immune system, caused by defects in the nicotinamide adenine dinucleotide phosphate oxidase enzyme complex present in phagocytic cells. These functional defects cause inability to produce reactive oxygen species in phagocytes, affecting the elimination of some pathogenic microorganisms within the phagolysosome. The diagnosis of this disease is currently made by means of the 1,2,3-dihydrorodamine test assisted by multiparametric flow cytometry, or the staining of phagocytes with nitro-blue tetrazolium assisted by light microscopy. Objectives: To characterize molecular and pathophysiologically the chronic granulomatous disease; and to discuss aspects related to the aforementioned diagnostic tests. Methods: In this work, a bibliographic-documentary research was carried out from scientific articles published from 1933 to 2018, for which the SciELO, PubMed and Springer databases were consulted. Development: The pathophysiological characteristics of chronic granulomatous disease are exposed, as well as the relationship between the most abundant genetic mutations in the affected population, and the severity of the clinical manifestations presented by the patients. In addition, the benefits and deficiencies of two techniques currently used to diagnose the disease are critically analyzed. Conclusions: Chronic granulomatous disease can generate severe immunological and inflammatory consequences, which are in line with the genetic characteristics expressed in the damaged enzyme complex. The diagnosis of the disease is more reliable, exhaustive and specific, using flow cytometry and its 1,2,3-dihydrorodamine test(AU)

Enfermedad granulomatosa crónica: diagnóstico mediante el ensayo de dihidrorodamina / Chronic granulomatous disease: diagnosis by the dihydrorhodamine assay

Resumen: Introducción: La enfermedad granulomatosa crónica (EGC) se caracteriza por una alteración de la función oxidativa de neutrófilos, presentando herencia ligada al cromosoma X (EGC LX) y autosómica recesiva (EGC AR). El ensayo de dihidrorodamina (DHR) es utilizado para el diagnóstico y detección de portadoras, además proporciona información sobre patrones de herencia. Objetivo: Detectar casos de EGC en niños con infecciones recurrentes y evaluar a sus familiares femeninos mediante el ensayo de DHR, para identificar portadoras y obtener información acerca de posibles patrones de herencia. Pacientes y Método: Fueron incluidos 107 pacientes (<18 años de edad) con sospecha clínica de EGC como neumonías, linfadenopatías y abscesos, remitidos por médicos de hospitales públicos, del 2014 al 2017. Además, se incluyeron seis mujeres, familiares de los niños con EGC. A las muestras de los pacientes se aplicó el ensayo DHR, expresando los resultados como índice de estimulación de neutrófilos (IE). Resultados: La mediana de edad de los pacientes fue de 3 años y 62/107 fueron varones. El IE promedio fue 39,7 ± 13,8 y 101/107 niños exhibieron un cambio completo de fluorescencia de DHR. En 2/107 niños no se observó dicho cambio (IE = 1,0), lo cual indica posible EGC LX, y un tercer niño mostró un leve cambio (IE = 4,8), compatible con EGC AR. En 5/6 mujeres se encontró un patrón bimodal, indicando un estado de portadora. Conclusiones: Fueron detectados tres casos de EGC y cinco portadoras mediante el ensayo de DHR, realizado por primera vez en Paraguay. También se obtuvo información sobre los posibles patrones de herencia, EGC LX en dos familias y un caso probable de EGC AR.

Abstract: Introduction: Chronic granulomatous disease (CGD) is characterized by an alteration of the neutrophil oxidative function. Its inheritance patterns are linked to the X chromosome (X-linked CGD) and autosomal recessive (AR CGD). The dihydrorhodamine (DHR) assay is used for the diagnosis and detection of carriers and provides information on inheritance patterns. Objective: To detect CGD cases in chil dren with recurrent infections and to evaluate their female relatives through the DHR assay to iden tify carriers and obtain information about possible inheritance patterns. Patients and Method: 107 patients (<18 years of age) with clinical suspicion of CGD such as pneumonia, lymphadenopathies, and abscesses were included, referred by physicians from public hospitals between 2014 and 2017. Six female relatives of children with CGD were also included. The DHR assay was performed on all patient samples and the results were expressed as neutrophils stimulation index (SI). Results: The median age of patients was 3 years and 62/107 of them were male. The average SI was 39.7±13.8 and a complete shift of DHR was found in 101/107 children. In 2/107 children, no DHR shift was observed (SI=1.0) indicating possible X-linked CGD, and a third child showed a slight DHR shift (SI=4.8) compatible with AR CGD. 5/6 female relatives presented a bimodal pattern, showing a carrier status. Conclusions: Three cases of CGD and five female carriers were detected through the DHR assay, being the first time that this technique was used in Paraguay. Information on the most likely inheri tance patterns, two X-linked CGD, and one AR CGD case was also obtained.

Función del sistema NADPH oxidasa en la formación de trampas extracelulares de los neutrófilos (NETs) / Role of nadph oxidase system in the neutrophil extracelular Traps (NETs) formation

Las trampas extracelulares de los neutrófilos son estructuras fundamentalmente compuestas de cromatina y proteínas granulares, que una vez liberadas constituyen un mecanismo de defensa que tiene la capacidad de atrapar y destruir microorganismos patógenos. El proceso que libera estas estructuras es conocido como NETosis y en el caso que provoque muerte celular, esta es diferente a la apoptosis y a la necrosis. Si bien no se conocen todos los eventos moleculares involucrados en la formación de las NETs, se sabe que dependiendo del estímulo, las especies reactivas del oxígeno son esenciales para que ocurra la descondensación de la cromatina y se lleve a cabo el proceso de NETosis(AU)

Neutrophil extracellular traps (NETs) are structures mainly composed of chromatin and granule proteins that once released constitute a defense mechanism due to their ability to trap and destroy pathogen microorganisms. The process by which these structures are released is known as NETosis and in case this may lead to cell death is different to apoptosis and necrosis. Although all the molecular events involved in the formation of NETs are poorly understood, it is known that depending on the stimulus, reactive oxygen species (ROS) are essential to the chromatin decondensation and subsequent NETs formation(AU)

Enfermedad granulomatosa crónica: tres casos clínicos con diferentes formas de presentación / Chronic granulomatous disease: three cases with different presentations

Introducción: La enfermedad granulomatosa crónica (EGC) es una forma infrecuente de inmunodeficiencia primaria que se caracteriza por una sensibilidad anormal a infecciones bacterianas y fúngicas, debida a un déficit en el complejo nicotinamida adenina dinucleótida fosfato oxidasa (NADPH) en los fagocitos. Objetivo: Describir tres casos de EGC con énfasis en su forma de presentación y realizar una revisión del tema. Casos Clínicos: Se presentan tres casos clínicos, dos de ellos con relación de parentesco (primos en primer grado). Se llegó a diagnóstico molecular en uno de los casos. Se destacan las manifestaciones clínicas: infecciones recurrentes, abscesos, adenitis y granulomas, y complicaciones, con la finalidad de facilitar la sospecha diagnóstica de EGC, debido a la importancia del diagnóstico temprano y el consejo genético. Conclusiones: La EGC es un trastorno inmunológico primario congénito infrecuente, con herencia ligada a X en su mayoría, pero también con formas autosómicas recesivas, con una forma de presentación característica y cuyo diagnóstico debe ser oportuno para evitar complicaciones, realizar profilaxis y tratamiento agresivo de las infecciones y consejo genético.

Introduction: Chronic granulomatous disease (CGD) is a rare form of primary immunodeficiency disease, characterized by an abnormal susceptibility to bacterial and fungal infections, and it is caused by a deficit in the phagocyte nicotinamide adenine dinucleotide phosphate oxidase complex (NADPH), resulting in the inability to generate reactive oxygen species that destroy micro-organisms. The diagnosis is based on clinical characteristics and analysis of phagocytes, and later confirmed by molecular studies. Its management should consider antimicrobial prophylaxis, a search for infections and aggressive management of these. Objective: To describe three cases of CGD emphasizing their forms of presentation and to conduct a review of the condition. Case reports: Three case reports, two of them first cousins, are presented. Molecular diagnosis was reached in one of the cases. Recurrent infections, abscesses, adenitis, granulomas and complications are identified to facilitate the suspected diagnosis of CGD, bearing in mind the importance of early diagnosis and genetic counseling. Conclusions: EGC is a rare congenital primary immunodeficiency disorder, mostly with X-linked inheritance, autosomal recessive form, and a specific presentation form. Its diagnosis should be timely to avoid complications. Prophylaxis and aggressive treatment of infections should be performed, as well as genetic counseling.

Chronic granulomatous disease: why an inflammatory disease?

Chronic granulomatous disease is a primary immunodeficiency caused by mutations in the genes encoding subunits of the phagocytic NADPH oxidase system. Patients can present with severe, recurrent infections and noninfectious conditions. Among the latter, inflammatory manifestations are predominant, especially granulomas and colitis. In this article, we systematically review the possible mechanisms of hyperinflammation in this rare primary immunodeficiency condition and their correlations with clinical aspects.

Caracterización clínica y genético-molecular de un paciente con enfermedad granulomatosa crónica ligada al X: reporte de una nueva mutación asociada al splicing: caso clínico / Clinical, genetic and molecular characterization of the X-linked chronic granulomatous disease: a case report of a new splicing mutation

Introducción: La Enfermedad Granulomatosa Crónica (EGC) se presenta como consecuencia de mutaciones en los genes que codifican 5 de las subunidades del sistema NADPH oxidasa humano. Su forma más común es causada por cambios en el gen CYBB que codifica gp91 phox. Objetivo: Identificar el defecto molecular que lleva a la presentación de la EGC. Caso clínico: Paciente de sexo masculino con antecedentes de enfermedad diarreica aguda y abscesos perianales recurrentes desde los 2 meses. A los 6 meses, presentó una inflamación crónica del colon y colitis bacteriana. A los 3 años tenía infecciones en las vías respiratorias inferiores y perianales. Estudio compatible con EGC. El análisis del ADNc identificó expresión anormal del ARNm, lo cual se confirmó al realizar la secuenciación. Específicamente se observó la ausencia del exón 2. Adicionalmente, los datos de la secuenciación del ADNg identificaron una alteración en el sitio aceptor de "splicing" del intrón 1, que incluye una deleción seguida de la inserción de 3 nucleótidos (c.46-14_-11delTTCT insGAA). Conclusiones: Se presenta el primer estudio molecular de un paciente con EGC por defecto de "splicing" reportado en Colombia. La definición de la mutación y su correlación con el fenotipo es importante para proveer una apropiada consejería genética al paciente y su familia.

Chronic granulomatous disease (CGD) is caused by mutations in the genes that encode five of the subunits of the human NADPH oxidase. The most common form is caused by mutations in CYBB, the human gene encoding gp 91 phox. Objective: To identify the molecular defects causing CGD. Case report: A male patient with a history of acute diarrhea and recurrent perianal abscess since two months old. At 6 months, the patient presented a chronic inflammatory disease of the colon and bacterial colitis. After three years, he developed infections in the lower and perianal respiratory tract. The cDNA analysis identified abnormal mRNA expression, which was confirmed by sequencing. Specifically the exclusion of exon 2 was observed. Additionally, gDNA sequencing identified an alteration in the acceptor splice site of intron 1, including a deletion followed by insertion of three nucleotides (c.46-14_-11delTTCT insGAA). Conclusions: The first molecular study of a patient with CGD due to splicing pattern change, reported in Colombia, is presented. The definition of the mutation and its correlation with the phenotype is essential to provide appropriate genetic counseling to patients and their families.

Genetic Analysis of 10 Unrelated Korean Families with p22-phox-deficient Chronic Granulomatous Disease: An Unusually Identical Mutation of the CYBA Gene on Jeju Island, Korea

Chronic granulomatous disease (CGD) is a rare hereditary disorder characterized by recurrent life-threatening bacterial and fungal infections. The underlying defect in CGD is an inability of phagocytes to produce reactive oxygen species as a result of defects in NADPH oxidase. Considering that CGD generally affects about 3-4 in 1,000,000 individuals, it is surprising that the prevalence of CGD on Jeju Island is 20.7 in 1,000,000 individuals. We performed genetic analysis on 12 patients from 10 unrelated families and found that all patients had an identical homozygous single-base substitution of C to T in exon 1 (c.7C>T) of the CYBA gene, which was expected to result in a nonsense mutation (p.Q3X). Because Jeju Island has long been a geologically isolated region, the high prevalence of CGD on Jeju Island is presumably associated with an identical mutation inherited from a common ancestor or proband.

Nonsense mutations of the CYBB gene in two Thai families with X-linked chronic granulomatous disease.

X-linked chronic granulomatous disease (X-CGD) is an immunodeficiency disorder characterized by defective intracellular killing of microorganisms due to the neutrophils' inability to generate superoxide ions. Although it is always caused by mutations in the CYBB gene, clinical and molecular characteristics vary in different ethnic backgrounds. Two unrelated Thai boys presented with severe persistent pulmonary infections at the age of two months. Their abnormal dihydrorhodamine (DHR) flow cytometry assays supported the diagnosis of X-CGD. Mutation analysis was performed by polymerase chain reaction (PCR) amplification and sequencing of the entire coding regions of CYBB. Mutations identified were confirmed by restriction enzyme analyses. PCR-sequencing of the entire coding regions of CYBB identified nonsense mutations, 271C>T (R91X) in exon 4 and 456T>A (Y152X) in exon 5, in probands of each family. Both of the patients' mothers were found to be carriers. This observation supports that CYBB is the gene responsible for X-CGD across different populations and nonsense mutations are associated with severe phenotypes.

Identificación de mutaciones en el gen CYBB que llevan al fenotipo de la enfermedad granulomatosa crónica ligada al cromosoma X: Reporte de una nueva mutación / Report of a new mutation in CYBB gene in two patients with X linked chronic granulomatous disease

Background: The X-linked form of chronic granulomatous disease (CGD) is a primary immunodeficiency that affects phagocytes of the innate immune system and is characterized by an increased susceptibility to severe bacterial and fungal infections. It is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of phagocyte NADPH oxidase. Aim: To identify the mutation in the CYBB gene in two unrelated patients from Chile with the diagnosis of X-linked CGD and their families. Patients and methods: The molecular genetic defects of two unrelated patients from Chile with X-linked CGD caused by defects in the CYBB gene were investigated. The underlying mutation was investigated by single strand conformation polymorphism (SSCP) analysis of PCR-amplified genomic DNA and by sequencing of the affected gene region. Results: We found an insertion c.1267_1268insA in exon 10 leading to a frameshift mutation. This mutation is a novel report. We also identified a splice site mutation in the other patient, that presented a c.1326 +1 G>A substitution in intron 10. The mutation was also detectable in his heterozygous mother. Conclusions: This is the first report of the clinical and molecular characterization of Chilean patients with mutations in CYBB gene.

Carrier screening and prenatal detection of chronic granulomatous disease in Iran

To develop an application that is simple and reliable using the nitroblue tetrazolium [NBT] method that clearly differentiates chronic granulomatous disease [CGD] patients with heterozygous carriers in groups suspected with CGD. This study was carried out in Shiraz University of Medical Sciences from October 2002 and March 2004. The study included 260 samples consisting of 123 children [2-24 months] and 106 neonates [<2 months], either suspected with bacterial infection or are immunodeficient, and 31 cord blood samples. Fifty healthy adult individuals were also diagnosed as normal control. Neutrophil reduction of NBT can be stimulated in vitro by protein kinase agonists such as phorbol myristate acetate [PMA], resulting to superoxide anion release. The PMA is an exceptionally powerful stimulant and when we used it in conjunction with adherence of glass slides, it causes transformation of nearly 100% of all normal neutrophils, and reduces NBT to formazan deposits. Of 260 blood samples, 12 unrelated CGD patients and 16 carriers of X-linked or autosomal recessive CGD patients were diagnosed. The carriers had a range of 15-75% stimulated neutrophils. We have established a PMA-stimulated NBT test for the detection of CGD patients, which clearly differentiate the CGD patients from heterozygote carriers. The results in the cord fetal blood samples indicate that this test may be used for antenatal diagnosis of affected boys, carrier females and autosomal recessive variants of CGD. The technique is simple, fast, inexpensive, and requires only a few microliters of blood

Espectro clínico e defeitos genético - moleculares de pacientes com doença granulomatosa / Clinical spectrum and molecular genetics of chronic granulomatous disease

Os fagócitos contém uma nicotinamida adenina dinucleotídeo fosfato (NADPH) oxidase associada à menbrana, a qual gera superóxido e outros reativos intermediários do oxigênio, os quais apresentam atividades microbicida, tumoricida e inflamatória. Defeitos nesta oxidase resultam na doença granulomatosa crônica (DGC), cujo quadro clínico caracteriza-se por infecções graves, recorrentes e de início precoce, o que demostra a relevância clínica deste sistema enzimático, como mecanismo de defesa. Temos como objetivo revisar as características clínicas e os defeitos genético-moleculares desses pacientes, incluindo a experiência brasileira e latino-americana.

Enfermedad granulomatosa crónica: aspectos actuales / Chronic granulomatous disease: current aspects

La enfermedad granulomatosa crónica (EGC) es una inmunodeficiencia primaria causada por mutaciones en los genes que codifican para cualquiera de las 4 subunidades que conforman a la enzima adenina dinucleótido fosfato oxidasa (NADPH oxidasa), encargada de regular la producción de especies oxidantes microbicidas que constituyen la primera vía de defensa del organismo contra los microorganismos infecciosos. Esta es una deficiencia muy heterogénea clasificada en EGC ligada al cromosoma X (subunidad gp91-phox) y autosómica recesiva, donde puede estar afectada cualquiera de las siguientes subunidades de la enzima: p22-phox, p47-phox y p67-phox. Dentro de los hallazgos clínicos más frecuentes se encuentran entre otros la linfadenopatía, hepatoesplenomegalia, neumonía. Para el tratamiento de la enfermedad se utilizan antibióticos de amplio espectro y el interferón gamma para el tratamiento de infecciones severas en la EGC ligada al cromosoma X. Actualmente se están realizando estudios para utilizar la terapia génica somática como posible cura de la enfermedad

Doença granulomatosa crônica autossômica: relato de caso e análise genético-molecular de dois irmãos brasileiros / Autosomal chronic granulomatous disease: case report and mutation analysis of two Brazilian siblings

OBJETIVO: Relatar dois casos de irmãos com doença granulomatosa crônica. A doença granulomatosa crônica é uma imunodeficiência primária caracterizada por atividade microbicida deficiente. Mutações no gene que codifica a proteína p47-phox (NCF-1) estão presentes em 30 por cento dos casos de doença granulomatosa crônica. Essa forma da doença é de herança autossômica recessiva e resulta em fenótipo de evolução mais benigno e início tardio em relação à forma ligada ao X, que corresponde a 56 por cento dos casos. DESCRIÇAO: Caso 1 - paciente feminina, iniciou infecções de repetição aos 10 anos, com impetigo, seguido de pneumonia grave 6 meses após. A gravidade da infecção pulmonar associada a abscesso hepático e sua refratariedade ao tratamento demandaram investigação laboratorial para imunodeficiência, com teste do nitroblue tetrazolium e dosagem de ânion superóxido compatíveis com doença granulomatosa crônica. A avaliação dos familiares confirmou o mesmo diagnóstico em seu irmão (Caso 2), que também iniciou infecções de repetição com impetigo aos 10 anos e pneumonia 6 meses após, porém tratada com sucesso ambulatorialmente. A análise de polimorfismo conformacional de cadeia simples revelou alteração da mobilidade eletroforética do éxon 2 do gene NCF-1. Identificou-se uma deleção dos nucleotídeos GT no éxon 2 por seqüenciamento do DNA. COMENTARIOS: Este estudo mostra a importância da avaliação de familiares, mesmo quando não apresentam história clínica típica de doença granulomatosa crônica. A identificação da mutação e sua correlação com o fenótipo dos pacientes é importante para estabelecer o prognóstico e o aconselhamento genético.

Doença granulomatosa crônica da infância: novos polimorfismos no gene NCF2 / Chronic granulomatous disease of childhood: new polymorphisms in NCF2 gene

Objetivos: Mutações no gene NCF2 resultam na forma autossônica recessiva da doença granulomatosa crônica da infância. Além de mutações conhecidas, descreveu-se em pacientes com doença granulomatosa crônica da infância duas novas substituições no gene NCF2. O objetivo deste estudo foi investigar se estas substituições constituem polimorfismos do gene NCF2. Métodos: Investigamos a freqüência de duas substituições na seqüência do gene NCF2 em 214 doadores sadios. A primeira é uma transição de C T na posição -23 da região 5' reguladora. A segunda é uma transição de A G na posição -21 da região 3' terminal do íntron 10. Extraímos DNA genômico de células do sangue periférico. O DNA foi amplificado por meio de PCR com primers específicos para o gene NCF2, analisado quanto à presença de polimorfismos conformacionais de cadeias simples, digerido com endonucleases específicas e sequenciado. O cálculo das freqüências genotípicas e alélicas seguiu a lei de Hardy e Weinberg. Resultados: Cem indivíduos foram avaliados quanto à presença da transição C T na posição -23 da região 5' reguladora; sendo 67 porcento homozigotos para o alelo c, 32 porcento heterozigotos, e apenas 1 porcento homozigoto para T. Cento e quatorze indivíduos foram analisados quanto à presença da transição A G na posição -21 da região 3' terminal do íntron 10; dos quais 36 porcento foram homozigotos para A, 43 porcento heterozigotos e 21 porcento homozigotos para G. Conclusão: Considerando as freqüências alélicas, concluímos que essas variantes correspondem a polimorfismos do gene NCF2. Suas possíveis implicações na expressão do gene NCF2 constituem objeto de pesquisa atual em nosso laboratório.

Molecular Analysis of X-linked Chronic Granulomatous Disease in Five Unrelated Korean Patients

Chronic granulomatous disease (CGD) is a fatal genetic disorder in which phagocytes fail to produce antimicrobial superoxide because of NADPH oxidase deficiency. Molecular defects in CYBB gene causing X-linked CGD are responsible for about 70% of all cases. This study was done to confirm genetic defects of CYBB gene in five Korean patients who were highly suggestive of having CGD by clinical history. We performed initial screening for five unrelated Korean patients using single strand conformation polymorphism (SSCP) and then selective sequencing for the regions involving the abnormal bands. Activated NBT tests revealed that all patients were X-linked. SSCP analysis for CYBB gene showed abnormal bands in all patients. The molecular defects of five patients were as follows: c.1663insT, c.1111-1G>T, c.39_40insG, c.927delC and c.434T>C mutation. This result will help the families with prenatal diagnosis or genetic counseling.

Caracterización clínico molecular de la enfermedad granulomatosa crónica autosómica recesiva causada por déficit de p47-phox / Clinical and molecular characterization af autosomal recessive chronic granulomatous disease caused by p47-phox deficiency

Background: The cytosolic protein p47-phox (phagocyte oxidase) is one of the essential components of the superoxide generating system in phagocytes and its defect causes approximately 30 percent of the chronic granulomatous disease (CGD) cases. Aim: Two patients were studied, belonging to the same family, without a consanguinous background, in which deficiency or absence of superoxide generation was found together with recurrent and severe infections in one case and benign infections in the second. Methods: The presence of gp91-, p67- and p47-phox in patients and controls was determined by Western Blot analysis of granulocytes. Sequencing of PCR amplified DNA was performed by an enzimatic method. Results: Western Blot analysis showed normal expression of gp91 and p67 and absence of p47-phox. The molecular genetic study demonstrated a homocygotic dinucleotide GT (GT) deletion at the beginning of exon 2 of the p47-phox gene. The same mutation has been found in European, American and Japanese patients. Conclusions: The molecular characterization of this pathology done for the first time in Chile is important for diagnostic classification, patient prognosis, and adequate genetic advice and a possible future therapy

Enfermedad granulomatosa cronica (sindrome de deficiencia de oxidasa fagocitaria): analisis de un paciente adulto y su familia / Chronic granulomatous disease (fagocytic oxydase deficiency syndrome): analysis of an adult patient and his family

La enfermedad granulomatosa cronica es un defecto congenito de los fagocitos, que se manifiesta clinicamente con infecciones severas y recurrentes principalmente en niños. La forma clasica debe sospecharse en los niños con infecciones recurrentes y severas en piel, pulmones y otros organos y la presencia de granulomas en esos sitios. No obstante, en los ultimos años, se ha descrito este problema en adultos, con una amplia gama de patrones geneticos y variadas manifestaciones clinicas. Se señala el de "sindrome de deficiencia de oxidasa fagocitaria" como el nombre mas adecuado, puesto que incluye todos los posibles defectos bioquimicos observados en estos pacientes. En el presente articulo se analizan los hallazgos en un paciente adulto con este trastorno y su familia.