title: The rs361525 polymorphism does not increase production of tumor necrosis factor alpha by monocytes from alpha-1 antitrypsin deficient subjects with chronic obstructive pulmonary disease - a pilot study.
abstract: BACKGROUND
  Polymorphisms in the TNF-A gene have been associated with chronic obstructive pulmonary disease (COPD) in some case-control studies. Previous work has shown that COPD/chronic bronchitis subjects with alpha-1 antitrypsin deficiency with the rs361525 TNF-α single nucleotide polymorphism have 100 times more TNF-in spontaneous sputum than disease matched controls. Our objective was to determine if the presence of this polymorphism increased TNF-α production by blood monocytes from COPD subjects.
  
  
  RESULTS
  Monocytes from 18 COPD/alpha-1 antitrypsin deficient subjects, with and without the rs361525 polymorphism, were cultured in the presence or absence of lipopolysaccharide. Cell-free supernatants were analyzed by ELISA and real-time PCR performed using cDNA from extracted RNA. Baseline expression of TNF-α messenger RNA was no different between the groups. No difference in messenger RNA or secreted protein was observed over time in un-stimulated cells. TNF-α messenger RNA expression and protein was not higher in lipopolysaccharide-stimulated monocytes from subjects with the polymorphism compared to cells from patients with the wild-type allele.
  
  
  CONCLUSIONS
  This small pilot study did not provide an explanation for the findings of earlier observations of the association of the rs361525 polymorphism with TNF-α in airways secretions. Possible reasons for the lack of concordance include the study of blood rather than tissue cells, the use of a single stimulant rather than biological secretions and the need for far greater subject numbers to overcome intra-subject variation in monocyte TNF-α production.
authors.affiliation: Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK. jenniegane@doctors.org.uk.|Centre for Translational Inflammation Research, New Queen Elizabeth Hospital, University of Birmingham Laboratories, Institute of Inflammation and Ageing, Edgbaston, Birmingham, B15 2TT, UK. jenniegane@doctors.org.uk.
authors.forename: Jennie M
authors.initials: JM
authors.lastname: Gane
authors.affiliation: Lung Investigation Unit, University Hospital Birmingham NHS Foundation Trust, Edgbaston, Birmingham, UK. Rob.Stockley@uhb.nhs.uk.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Stockley
authors.affiliation: Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK. e.sapey@bham.ac.uk.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Sapey
mesh_terms: D000368:Aged; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000378:metabolism; D010865:Pilot Projects; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D029424:Pulmonary Disease, Chronic Obstructive / Q000453:epidemiology / Q000235:genetics* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics*; D019896:alpha 1-Antitrypsin Deficiency / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics* / Q000378:metabolism
chemical_list: D014409:Tumor Necrosis Factor-alpha