title: In vitro allergen challenge of peripheral blood induces differential gene expression in mononuclear cells of asthmatic patients: inhibition of cytosolic phospholipase A2alpha overcomes the asthma-associated response.
abstract: BACKGROUND
  Existing treatments for asthma are not effective in all patients and disease exacerbations are common, highlighting the need for increased understanding of disease mechanisms and novel treatment strategies. The leukotriene pathway including the enzyme responsible for arachidonic acid release from cellular phospholipids, cPLA(2)alpha, is a major contributor to asthmatic responses and an attractive target in asthma therapies.
  
  
  OBJECTIVE
  The study reported here investigates (a) the differential effects of in vitro exposure of peripheral blood mononuclear cells (PBMCs) to allergen between asthma and healthy subjects, and (b) the contribution of cPLA(2)alpha to these differences in gene expression.
  
  
  METHODS
  In vitro responses of asthma (N=26) and healthy (N=11) subject PBMC samples to allergen stimulation in the presence and absence of cPLA(2)alpha inhibition or 5-lipoxygenase inhibition were compared at the gene expression level using oligonucleotide arrays and at the protein level using ELISA.
  
  
  RESULTS
  Subject samples within both asthma and healthy groups showed allergen-dependent cytokine production and allergen-dependent gene expression changes, although transcriptional profiling identified 153 genes that were modulated significantly differently by allergen between asthma and healthy subjects. Among these were genes previously associated with asthma, but the majority (about 80%) have not previously been associated with asthma.
  
  
  CONCLUSIONS
  Transcriptional profiling elucidated novel gene expression differences between the asthmatic and healthy subject samples. Although 5-lipoxygenase inhibition did not significantly affect allergen-modulated gene expression, the inhibition of cPLA(2)alpha activity affected many of the allergen-dependent, asthma-associated gene expression changes.
authors.affiliation: Wyeth Research, Cambridge, MA 02140, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Whalen
authors.forename: H
authors.initials: H
authors.lastname: Legault
authors.forename: C
authors.initials: C
authors.lastname: Hang
authors.forename: A
authors.initials: A
authors.lastname: Hill
authors.forename: M
authors.initials: M
authors.lastname: Kasaian
authors.forename: D
authors.initials: D
authors.lastname: Donaldson
authors.forename: G W
authors.initials: GW
authors.lastname: Bensch
authors.forename: G
authors.initials: G
authors.lastname: Bensch
authors.forename: J
authors.initials: J
authors.lastname: Baker
authors.forename: P S
authors.initials: PS
authors.lastname: Reddy
authors.forename: N
authors.initials: N
authors.lastname: Wood
authors.forename: M K
authors.initials: MK
authors.lastname: Ramarao
authors.forename: D K
authors.initials: DK
authors.lastname: Ellis
authors.forename: C
authors.initials: C
authors.lastname: Csimma
authors.forename: C
authors.initials: C
authors.lastname: McKee
authors.forename: J D
authors.initials: JD
authors.lastname: Clark
authors.forename: J
authors.initials: J
authors.lastname: Ryan
authors.forename: A J
authors.initials: AJ
authors.lastname: Dorner
authors.forename: M
authors.initials: M
authors.lastname: O'Toole
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology* / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism; D001249:Asthma / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D001565:Benzoates / Q000494:pharmacology; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D054513:Group IV Phospholipases A2 / Q000037:antagonists & inhibitors* / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000201:enzymology / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: C531069:4-(3-(5-chloro-2-(2-(((2,6-dimethylbenzyl)sulfonyl)amino)ethyl)-1-(diphenylmethyl)-1H-indol-3-yl)propyl)benzoic acid; D000485:Allergens; D001565:Benzoates; D016207:Cytokines; D013449:Sulfonamides; D016718:Arachidonic Acid; D054513:Group IV Phospholipases A2