RESUMO
PURPOSE: Chronic granulomatous disease (CGD) is an inherited disorder, with phagocytes failing to produce antimicrobial superoxide due to deficient NADPH oxidase activity. Mutations in the gene encoding CYBB are responsible for the majority of the CGD cases. To date, there have been no reports on large samples of children with CGD in China. Therefore, in this study, we described the clinical and molecular features of 38 suspected CGD patients from 36 unrelated Chinese families. METHODS: Clinical diagnosis was performed using dihydrorhodamine assays detected by flow cytometry. Molecular analysis was used to identify underlying CGD-causative genes. RESULTS: The mean age of onset in our 38 patients was 3.4 months, while the mean age at diagnosis was 31.7 months. Apart from recurrent pneumonia and abscesses, tuberculosis (TB) and Bacille Calmette-Guerin (BCG) infections were notable features in our cohort. Overall, 17 cases died and patient 1 did not participate in the follow-up period . In total, we identified 29 different CYBB gene mutations in 31 patients. We found NCF1 and CYBA mutations in 3 and 2 patients, respectively. In addition, we identified 31 carriers and prenatally diagnosed 4 CGD and 4 healthy fetuses. CONCLUSIONS: The results of our study demonstrate that children with BCG infections or recurrent TB infections should have immune function screening tests performed. Moreover, newborns with family histories of primary immunodeficiency diseases should avoid of BCG vaccination. Molecular analysis is an important tool for identifying patients, carriers, and high-risk CGD fetuses.
Assuntos
Doença Granulomatosa Crônica/epidemiologia , Glicoproteínas de Membrana/metabolismo , NADPH Oxidases/metabolismo , Tuberculose Pulmonar/epidemiologia , Idade de Início , Criança , Pré-Escolar , China , Análise Mutacional de DNA , Feminino , Seguimentos , Doença Granulomatosa Crônica/diagnóstico , Humanos , Lactente , Recém-Nascido , Masculino , Glicoproteínas de Membrana/genética , Mutação/genética , NADPH Oxidase 2 , NADPH Oxidases/genética , Gravidez , Risco , Tuberculose Pulmonar/diagnósticoRESUMO
Type 1 hyper-IgM syndrome (HIGM1) is a rare primary immunodeficiency disease caused by mutations in the CD40L gene. Patients often present with recurrent infections and autoimmune manifestations. We investigated the clinical and molecular characteristics of HIGM1 in thirteen patients from the Chinese mainland and examined the proportion of CD4(+)CD25(+)FoxP3(+)Treg, Th17, and Th1 cells in the peripheral blood. We identified ten distinct CD40L mutations in eleven patients: one missense mutation, one nonsense mutation, one insertion mutation (in frame), and seven deletions. Six of these mutations were novel. We observed the percentage of Tregs in the peripheral blood of HIGM1 patients decreased markedly compared with that in healthy controls, but no statistically significant differences was found in the percentages of Th17 and Th1. The identified mutations reflect the heterogeneity of the CD40L gene in HIGM1. Precise genetic diagnosis of HIGM1 will enable appropriate therapeutic interventions, reliable detection of carriers, and genetic counseling. Skewed Treg, Th17/Treg, and Th1/Treg profiles may be associated with immune responses to autoimmunity or infection, which requires replication in larger studies.
Assuntos
Ligante de CD40/genética , Síndrome de Imunodeficiência com Hiper-IgM/genética , Mutação , Linfócitos T Reguladores/imunologia , Povo Asiático , Sequência de Bases , Antígenos CD4/genética , Antígenos CD4/imunologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Estudos de Coortes , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/imunologia , Expressão Gênica , Humanos , Síndrome de Imunodeficiência com Hiper-IgM/etnologia , Síndrome de Imunodeficiência com Hiper-IgM/imunologia , Síndrome de Imunodeficiência com Hiper-IgM/patologia , Imunofenotipagem , Lactente , Subunidade alfa de Receptor de Interleucina-2/genética , Subunidade alfa de Receptor de Interleucina-2/imunologia , Masculino , Dados de Sequência Molecular , Linfócitos T Reguladores/patologia , Células Th1/imunologia , Células Th1/patologia , Células Th17/imunologia , Células Th17/patologiaRESUMO
METHODS: Two hundred three children with genetically proven primary immunodeficiency diseases (PIDs) from 197 unrelated families were enrolled from January 2005 to December 2011. RESULTS: On the basis of criteria developed by the International Union of Immunological Societies, 79 patients were diagnosed as "other well-defined immunodeficiency syndromes" (38.9%), 62 (30.6%) with "predominant antibody deficiencies," 26 (12.8%) with "congenital defects of phagocyte," 25 (12.3%) with "T- and B-cell immunodeficiency" and 11 (5.4%) with "diseases of immune dysregulation." The median time to the diagnosis was 27.9 months and the patients had a wide range of clinical presentations. In addition, a total of 23 pathogenic genes were identified and 213 mutations were detected, including 42 novel mutations. CONCLUSIONS: With the increase in the awareness of PIDs and diagnostic competence, more PID patients will be diagnosed and we will be able to more accurately identify the frequency and the distribution of PIDs in the most populous country in the world.
Assuntos
Síndromes de Imunodeficiência/genética , Síndromes de Imunodeficiência/patologia , Criança , Pré-Escolar , China/epidemiologia , Análise Mutacional de DNA , Feminino , Humanos , Síndromes de Imunodeficiência/terapia , Lactente , Masculino , Estudos RetrospectivosRESUMO
UNLABELLED: X-linked severe combined immunodeficiency (X-SCID) is a rare, life-threatening immune disorder, caused by mutations of the gene for the γ-chain (γc) of the interleukin-2 receptor, IL2RG. We analyzed the clinical, immunologic, and molecular characteristics of children with X-SCID, attempting to improve the diagnosis and treatment of X-SCID in China. METHODS: X-SCID was suspected in male infants with recurrent or persistent infections. Eleven male infants from ten unrelated Chinese families were included. The IL2RG gene was amplified and sequenced, followed by mutation analysis in these children and their female relatives. X-linked short tandem repeat (X-STR) typing was done to define the maternal lymphocyte engraftment. RESULTS: The 11 children exhibited recurrent infections and 10 of them had lymphopenia. B cells were present in all patients, T cells were markedly reduced in 10, and NK cells were markedly reduced in 9. Nine IL2RG gene mutations were identified in the 11 children, with 5 novel mutations. One patient was found to have the maternal lymphocyte engraftment. CONCLUSION: The clinical presentations and immunologic characteristics of the X-SCID patients were accordingly quite uniform despite the heterogeneity of mutations locating almost in the entire γc gene.
Assuntos
Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/diagnóstico , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genética , China , Análise Mutacional de DNA , Feminino , Humanos , Lactente , Recém-Nascido , MasculinoRESUMO
Respiratory syncytial virus (RSV) is the most common pathogen responsible for lower respiratory diseases in children. So far, there is no effective treatment or preventative vaccine available for RSV infection, although ribavirin and dexamethasone are commonly prescribed. Resveratrol has been shown to inhibit the replication of several other viruses, thus the effect of resveratrol on RSV-induced inflammatory mediators in 9HTEo cell cultures was evaluated, and possible mechanisms of action were explored and compared with dexamethasone and ribavirin. Incubation with resveratrol resulted in decreased IL-6 production and partial inhibition of RSV replication. Resveratrol treatment also inhibited virus-induced TIR-domain-containing adapter-inducing interferon-ß (TRIF) and TANK binding kinase 1 (TBK1) protein expression. These data demonstrate the ability of resveratrol to inhibit cytokine production by RSV in airway epithelial cells, indicating that it might be a therapeutic agent with both anti-inflammatory and antiviral potential for the treatment of RSV infection.
Assuntos
Proteínas Adaptadoras de Transporte Vesicular/biossíntese , Interleucina-6/biossíntese , Vírus Sinciciais Respiratórios/efeitos dos fármacos , Estilbenos/farmacologia , Replicação Viral/efeitos dos fármacos , Antivirais/farmacologia , Linhagem Celular , Dexametasona/farmacologia , Regulação para Baixo , Células Epiteliais/imunologia , Células Epiteliais/virologia , Humanos , Interleucina-8/biossíntese , Proteínas Serina-Treonina Quinases/biossíntese , Mucosa Respiratória/metabolismo , Mucosa Respiratória/virologia , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/imunologia , Vírus Sinciciais Respiratórios/fisiologia , Sistema Respiratório/metabolismo , Sistema Respiratório/virologia , Resveratrol , Ribavirina/farmacologiaRESUMO
OBJECTIVE: 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. METHOD: 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. RESULT: 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. CONCLUSION: 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.
Assuntos
Amniocentese , Doenças Fetais/diagnóstico , Diagnóstico Pré-Natal , Síndrome de Wiskott-Aldrich/diagnóstico , Feminino , Citometria de Fluxo , Humanos , Masculino , Gravidez , Síndrome de Wiskott-Aldrich/genética , Proteína da Síndrome de Wiskott-Aldrich/sangue , Proteína da Síndrome de Wiskott-Aldrich/genéticaAssuntos
Documentação , Pediatria , Criança , Documentação/normas , Humanos , Pediatria/normas , Guias de Prática Clínica como AssuntoRESUMO
Omenn syndrome (OS) is a rare autosomal recessive genetic disorder and presents symptoms of severe combined immunodeficiency characterized by erythrodermia, eosinophilia, hepatosplenomegaly, lymphadenopathy, and elevated serum IgE levels. OS has been found to be caused by mutations in RAG1 or RAG2 gene that result in partial V(D)J recombination activity. No study on OS has been reported in Chinese children so far. In this study, the genotype and phenotypes of three infants with OS from three unrelated Chinese families were investigated. All the three children had most of the characteristics of OS except normal serum IgE level. Compound heterozygosity mutations in RAG1 gene (1983 G>A; 2444 C>T and 2219 C>T; 3127 C>G) were identified in two cases, and a homozygous deletion mutation with a premature stop codon was found at residue 2302 of RAG1 gene (2302delT, I729X) in the remaining case, including three novel mutations (2302delT, I729X; 2219 C>T, R699W; and 3127 C>G, Y1001X). Spectratyping analysis of T-cell receptor ß-chain variable region (TCRVß) gene rearrangement was performed in case 1 and case 2. All the 25 TCRVß subfamilies presented monoclonal or oligoclonal peaks in case 1 and 11 TCRVß subfamilies were very weak or even absent in case 2. This was the first report about OS in Chinese children. Molecular genetic testing represents an important tool for early confirmed diagnosis and may allow accurate carrier detection and prenatal diagnosis.
Assuntos
Genes RAG-1/genética , Mutação , Imunodeficiência Combinada Severa/genética , Imunodeficiência Combinada Severa/fisiopatologia , Povo Asiático/genética , Análise Mutacional de DNA , Proteínas de Ligação a DNA/genética , Evolução Fatal , Genótipo , Heterozigoto , Proteínas de Homeodomínio/genética , Homozigoto , Humanos , Lactente , Recém-Nascido , Masculino , Proteínas Nucleares/genética , Fenótipo , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Imunodeficiência Combinada Severa/diagnóstico , Imunodeficiência Combinada Severa/patologiaRESUMO
AIM: To compare the inflammation between wild BALB/c mice and mice with severe combined immunodeficiency (SCID) with endotoxemia induced by lipopolysaccharide (LPS). METHODS: Endotoxemia models of wild and SCID mice were established by injecting LPS intraperitoneally. Serum was taken before and 3 h, 6 h, 12 h after injecting LPS, liver and lung were taken 12 h after injecting LPS. Alanine transarninase (ALT), aspartate aminotransferase (AST) and blood urea nitrogen (BUN) levels were measured by automatic biochemical analyzer. Liver and lung inflammation injury were observed by H.E staining. TNF-α, IFN-γ, IL-6 and MCP-1 levels in serum were detected by Cytometric Bead Array (CBA). RESULTS: (1) All of SCID mice (8/8) were dead at 12-24 h after injecting LPS, and only one BALB/c mouse (1/8) was dead. (2) ALT and AST levels of SCID mice 12 h after injecting LPS were higer than those of BALB/c mice(P<0.05), but there was no difference of BUN levels between them. (3) The blind liver and lung pathology scores of SCID mice were higher than those of BALB/c mice (P<0.05). (4) TNF-α, IFN-γ, IL-6 and MCP-1 levels in serum at 3 h, 6 h, 12 h after injecting LPS increased significantly, and the cytokine levels of SCID mice were higher than those of BALB/c mice(P<0.05). CONCLUSION: SCID mice don't only excessively secrete inflammatory cytokines after injecting LPS, but also lead to more severe endotoxemia and inflammation injury of organs, which are the important cause of mouse death. The above results also show that the adjustment deficiency of adaptive immunoresponse, abnormal augmentation of innate immunoresponse may be an important cause of severe SIRS of endangering life.
Assuntos
Endotoxemia/patologia , Inflamação/patologia , Alanina Transaminase/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Nitrogênio da Ureia Sanguínea , Quimiocina CCL2/sangue , Endotoxemia/induzido quimicamente , Endotoxemia/metabolismo , Inflamação/induzido quimicamente , Inflamação/imunologia , Inflamação/metabolismo , Interferon gama/sangue , Interleucina-6/sangue , Lipopolissacarídeos , Fígado/patologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Fator de Necrose Tumoral alfa/sangueRESUMO
Severe combined immunodeficiencies (SCID) are a group of rare inherited disorders with profound defects in T cell and B cell immunity. From 2005 to 2010, our unit performed testing for IL2RG, JAK3, IL7R, RAG1, RAG2, DCLRE1C, LIG4, AK2, and ZAP70 mutations in 42 Chinese and Southeast Asian infants with SCID adopting a candidate gene approach, based on patient's gender, immune phenotype, and inheritance pattern. Mutations were identified in 26 patients, including IL2RG (n = 19), IL7R (n = 2), JAK3 (n = 2), RAG1 (n = 1), RAG2 (n = 1), and DCLRE1C (n = 1). Among 12 patients who underwent hematopoietic stem cell transplantation, eight patients survived. Complications and morbidities during transplant period were significant, especially disseminated bacillus Calmette-Guérin disease which was often difficult to control. This is the first cohort study on SCID in the Chinese and Southeast Asian population, based on a multi-centered collaborative research network. The foremost issue is service provision for early detection, diagnosis, management, and definitive treatment for patients with SCID. National management guidelines for SCID should be established, and research into an efficient platform for genetic diagnosis is needed.
Assuntos
Mutação/genética , Imunodeficiência Combinada Severa/diagnóstico , Imunodeficiência Combinada Severa/genética , Agamaglobulinemia/etiologia , Agamaglobulinemia/imunologia , Povo Asiático/genética , Pré-Escolar , Estudos de Coortes , Proteínas de Ligação a DNA/genética , Endonucleases , Feminino , Transplante de Células-Tronco Hematopoéticas , Proteínas de Homeodomínio/genética , Humanos , Lactente , Recém-Nascido , Infecções/etiologia , Subunidade gama Comum de Receptores de Interleucina/genética , Janus Quinase 3/genética , Leucopenia/etiologia , Leucopenia/imunologia , Masculino , Proteínas Nucleares/genética , Receptores de Interleucina-7/genética , Imunodeficiência Combinada Severa/complicações , Imunodeficiência Combinada Severa/terapia , Resultado do TratamentoRESUMO
OBJECTIVE: Mutation in the heavy chain micro (microHC) gene causes a rare type of autosomal recessive agammaglobulinemia. Here we report the molecular and clinical characterization of a compound heterozygous mutation in the microHC gene in a patient with autosomal recessive agammaglobulinemia firstly from China. METHOD: A one-year and ten-month-old male patient and his parents were enrolled in this study. No mutation was found in BTK gene. The microHC gene of the patient and his parents were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the microHC transcripts. Sequencing was performed directly on the PCR products bidirectionally. RESULTS: Since 8 months of age, the patient had had recurrent fever and persistent cough. He suffered an acute right hemiplegia at 11 months of age and swelling and pain of left hip joint and right knee joint at one year and eight months of age. Cerebrospinal fluid routine examination showed that total cell count was 18 x 10(6)/L [normal range (0 - 15) x 10(6)/L], leukocyte count 7 x 10(6)/L [(0 - 15) x 10(6)/L] and biochemical examination showed protein 0.14 g/L (0.15 - 0.45 g/L), glucose 4.68 mmol/L (2.44 - 4.44 mmol/L) and chloride 116.3 mmol/L (120 - 132 mmol/L). Mycobacterium bovis was identified negative by cerebrospinal fluid smear examination. No obvious abnormity was detected on skull CT examination. Hydrothorax examination showed that total cell count was 848 x 10(6)/L, leukocyte count 785 x 10(6)/L and protein 30.8 g/L (< 30 g/L). Poliovirus isolation from stool sample of the patient was negative. The serum immunoglobulin (Ig) profile was IgG 181 mg/L (normal range, 5.09 - 10.09 g/L); IgM 28.8 mg/dl (400 - 1260 mg/dl) and IgA 22 mg/dl (310 - 670 mg/dl), IgE 4.6 U/ml (normal range < 150 U/ml). There were no B-cells but normal percentage of T-cells (67%) and NK cells (32%) were present in the peripheral blood. The patient had a compound heterozygous mutation in the microHC gene:on one allele, there was an alternative splice site mutation in exon 4 (1956 G > A), for which the patient's father was a carrier. Whereas on the other allele, an insert mutation between 170 and 175 in exon 1 with a premature stop codon (170 - 175 insert C, L11fs60X) was identified, for which the patient's mother was a carrier. The insert mutation in exon 1 of microHC gene was firstly reported. To detect the effect of the splice site mutation in exon 4, microHC cDNA of the patient was amplified by semi-nested PCR. The PCR products were purified and sequenced directly. A 136 bp of intron 4 was found in the transcripts and only the secreted isoform with a missense substitution is present in the patient, while synthesis of the membrane isoform is completely abolished. CONCLUSION: This is the first case with autosomal recessive agammaglobulinemia with compound heterozygous mutation in the microHC gene reported from China. A novel mutation in exon 1 was found.
Assuntos
Agamaglobulinemia/genética , Cadeias mu de Imunoglobulina/genética , Mutação , Agamaglobulinemia/congênito , Povo Asiático/genética , Sequência de Bases , DNA/genética , DNA Complementar/genética , Éxons , Heterozigoto , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Reação em Cadeia da PolimeraseRESUMO
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.