title: Functional effects of NAD(P)H oxidase p22(phox) C242T mutation in human leukocytes and association with thrombotic cerebral infarction.
abstract: BACKGROUND
  Previous study showed that polymorphism of the NAD(P)H oxidase p22(phox) gene is associated with atherosclerosis, although others could not confirm such association. We investigated the association between p22(phox) C242T polymorphism and thrombotic cerebral infarction and the role of this polymorphism on superoxide-production activity in human neutrophils and promyelocytic HL-60 cells as a model system.
  
  
  METHODS
  PCR-RFLP analysis revealed that genotype and allele frequencies of C242T polymorphism in 120 patients with thrombotic cerebral infarction and 177 control subjects. The superoxide-production activity in neutrophils was determined by cytochrome c reduction assay. To clarify the role of p22(phox) C242T polymorphism on NAD(P)H oxidase activity, we used transgenic HL-60 cells transfected with expression plasmids carrying p22(phox) cDNAs with or without C242T polymorphism.
  
  
  RESULTS
  Genotype and allele frequencies of C242T polymorphism in patients and control subjects were not significantly different. The superoxide-production activity in neutrophils with T allele was higher than in neutrophils without T allele. Moreover, expression analysis showed that superoxide-production activity in p22(phox) C242T-expressing HL-60 cells were significantly higher than in p22(phox)-expressing HL-60 cells.
  
  
  CONCLUSIONS
  We conclude that C242T of p22(phox) gene is not involved in thrombotic cerebral infarction but more likely in increased NAD(P)H oxidase activity in phagocytes.
authors.affiliation: Department of Neurology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Shimo-Nakanishi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hasebe
authors.forename: Asuka
authors.initials: A
authors.lastname: Suzuki
authors.forename: Hideki
authors.initials: H
authors.lastname: Mochizuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Nomiyama
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Isao
authors.initials: I
authors.lastname: Nagaoka
authors.forename: Yoshikuni
authors.initials: Y
authors.lastname: Mizuno
authors.forename: Takao
authors.initials: T
authors.lastname: Urabe
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution; D015152:Blotting, Northern; D002544:Cerebral Infarction / Q000209:etiology / Q000235:genetics*; D045304:Cytochromes c / Q000378:metabolism; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D018922:HL-60 Cells; D006801:Humans; D015151:Immunoblotting; D020767:Intracranial Thrombosis / Q000150:complications / Q000201:enzymology / Q000235:genetics*; D007962:Leukocytes / Q000378:metabolism*; D008297:Male; D026901:Membrane Transport Proteins / Q000235:genetics*; D008875:Middle Aged; D009154:Mutation; D009252:NADPH Dehydrogenase / Q000235:genetics*; D019255:NADPH Oxidases / Q000235:genetics*; D009504:Neutrophils / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D013481:Superoxides / Q000378:metabolism; D014162:Transfection
chemical_list: D026901:Membrane Transport Proteins; D010750:Phosphoproteins; D013481:Superoxides; D045304:Cytochromes c; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase