Hyperresponsiveness of neutrophils from gp 91(phox) deficient patients to lipopolysaccharide and serum amyloid A

We demonstrate here that neutrophils from chronic granulomatous disease (CGD) patients release larger amounts of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) than neutrophils from control subjects. Incremental cytokine production was observed under both basal and stimulated conditions in neutrophils from two CGD (gp 91(phox)) patients. The basal production of IL-8 was over seven-fold greater in CGD patients. The two samples assayed showed 3- and 10-fold increases in TNF-alpha. Basically, the same magnitude of increment was observed in lypopolysaccharide (LPS) and serum amyloid A protein (SAA)-stimulated cells. We also found that the levels of SAA and IL-8 were higher in the serum of CGD patients than the levels found in the serum of healthy donors. The increased responsiveness of neutrophils from CGD patients may be closely related with a deficiency in the assembly of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme system, or it may be due to a frequent inflammatory condition in these patients. In the latter case, the increased serum levels of systemic inflammatory factors, among them SAA, would contribute to the sustained accumulation and activation of phagocytes. Whatever the origin, the excessive production of cytokines may lead to inappropriate activation and tissue injury and even to increased susceptibility to invasive microorganisms, impairing the quality life of CGD patients. (C) 2004 Elsevier B.V. All rights reserved.9441671434

Hyperresponsiveness of neutrophils from gp 91(phox) deficient patients to lipopolysaccharide and serum amyloid A

We demonstrate here that neutrophils from chronic granulomatous disease (CGD) patients release larger amounts of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) than neutrophils from control subjects. Incremental cytokine production was observed under both basal and stimulated conditions in neutrophils from two CGD (gp 91(phox)) patients. the basal production of IL-8 was over seven-fold greater in CGD patients. the two samples assayed showed 3- and 10-fold increases in TNF-alpha. Basically, the same magnitude of increment was observed in lypopolysaccharide (LPS) and serum amyloid A protein (SAA)-stimulated cells. We also found that the levels of SAA and IL-8 were higher in the serum of CGD patients than the levels found in the serum of healthy donors. the increased responsiveness of neutrophils from CGD patients may be closely related with a deficiency in the assembly of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme system, or it may be due to a frequent inflammatory condition in these patients. in the latter case, the increased serum levels of systemic inflammatory factors, among them SAA, would contribute to the sustained accumulation and activation of phagocytes. Whatever the origin, the excessive production of cytokines may lead to inappropriate activation and tissue injury and even to increased susceptibility to invasive microorganisms, impairing the quality life of CGD patients. (C) 2004 Elsevier B.V. All rights reserved.Univ São Paulo, Fac Pharmaceut Sci, Dept Clin & Toxicol Analyses, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pediat, Paulista Sch Med, São Paulo, BrazilState Univ Campinas, Sch Med, Ctr Invest Pediat, Campinas, BrazilState Univ Campinas, Sch Med, Dept Pediat, Campinas, BrazilUniversidade Federal de São Paulo, Dept Pediat, Paulista Sch Med, São Paulo, BrazilWeb of Scienc

Independent mutational events are rare in the ATM gene: Haplotype prescreening enhances mutation detection rate

Mutations in the ATM gene are responsible for the autosomal recessive disorder ataxia,telangiectasia (A-T). Many different mutations have been identified using various techniques, with detection efficiencies ranging from 57 to 85%. in this study, we employed short tandem repeat (STR) haplotypes to enhance mutation identification in 55 unrelated A-T families of Iberian origin (20 Spanish, 17 Brazilian, and 18 Hispanic-American); we were able to identify 95% of the expected mutations. Allelic sizes were standardized based on a reference sample (CEPH 1347-2). Subsequent mutation screening was performed by PTT, SSCP, and DHPLC, and abnormal regions were sequenced. Many STR haplotypes were found within each population and six haplotypes were observed across several of these populations. Single nucleotide polymorphism (SNP) haplotypes further suggested that most of these common mutations are ancestrally related, and not hot spots. However, two mutations (8977C>T and 8264_8268delATAAG) may indeed be recurring mutational events. Common haplotypes were present in 13 of 20 Spanish A-T families (65%), in 11 of 17 Brazilian A-T families (65%), and, in contrast, in only eight of 18 Hispanic American families (44%). Three mutations were identified that would be missed by conventional screening strategies. in all, 62 different mutations (28 not previously reported) were identified and their associated haplotypes defined, thereby establishing a new database for Iberian A-T families, and extending the spectrum of worldwide ATM mutations. (C) 2003 Wiley-Liss, Inc.David Geffen Sch Med, Dept Pathol & Lab Med, Los Angeles, CA 90095 USAUFRJ, Inst Biofis Carlos Chagas Filho, Rio de Janeiro, BrazilStanford Univ, Sch Med, Stanford Genome Technol Ctr, Palo Alto, CA USAHosp St Joan de Deu, Barcelona, SpainUNIFESP, Escola Paulista Med, Dept Pediat, São Paulo, BrazilNatl Childrens Hosp Dr Carlos Saenz Herrera, Serv Immunol, San Jose, Costa RicaHosp La Paz, Immunol Unit, Madrid, SpainUNIFESP, Escola Paulista Med, Dept Pediat, São Paulo, BrazilWeb of Scienc