The effect of lipoprotein-associated phospholipase A2 deficiency on pulmonary allergic responses in aspergillus fumigatus sensitized mice

Abstract

Background: Lipoprotein-associated phospholipase A₂(Lp-PLA₂)/platelet-activating factor acetylhydrolase (PAF-AH) has been implicated in the pathogenesis of cardiovascular disease. A therapeutic targeting of this enzyme was challenged by the concern that increased circulating platelet activating factor (PAF) may predispose to or increase the severity of the allergic airway response. The aim of this study was to investigate whether Lp-PLA₂gene deficiency increases the risk of PAF and IgE-mediated inflammatory responsesin vitroandin vivousing mouse models.Methods: Lp-PLA₂-/- mice were generated and back crossed to the C57BL/6 background. PAF-AH activity was measured using a hydrolysis assay in serum and bronchoalveolar lavage (BAL) samples obtained from mice.Aspergillus fumigatus(Af)-specific serum was prepared for passive allergic sensitization of micein vivoand mast cellsin vitro. β- hexosaminidase release was studied in bone marrow derived mast cells sensitized withAf-specific serum or DNP-IgE and challenged withAfor DNP, respectively. Mice were treated with lipopolysaccharide (LPS) and PAF intratracheally and studied 24 hours later. Mice were sensitized either passively or actively againstAfand were studied 48 hours after a single intranasalAfchallenge. Airway responsiveness to methacholine, inflammatory cell influx in the lung tissue and BAL, immunoglobulin (ELISA) and cytokine (Luminex) profiles were compared between the wild type (WT) and Lp-PLA₂-/- mice.Results: PAF-AH activity was reduced but not completely abolished in Lp-PLA₂-/- serum or byin vitrotreatment of serum samples with a high saturating concentration of the selective Lp-PLA₂inhibitor, SB-435495. PAF inhalation significantly enhanced airway inflammation of LPS treated WT and Lp-PLA₂-/- mice to a similar extent. Sensitized WT and Lp-PLA₂-/- bone-marrow derived mast cells released β-hexosaminidase following stimulation by allergen or IgE crosslinking to equivalent levels. Wild type and Lp-PLA₂-/- mice responded to passive or active allergic sensitization by significant IgE production, airway inflammation and hyperresponsiveness afterAfchallenge. BAL cell influx was not different between these strains while IL-4, IL-5, IL-6 and eotaxin release was attenuated in Lp-PLA₂-/- mice. There were no differences in the amount of total IgE levels in theAfsensitized WT and Lp-PLA₂-/- mice.Conclusions: We conclude that Lp-PLA₂deficiency in C57BL/6 mice did not result in a heightened airway inflammation or hyperresponsiveness after PAF/LPS treatment or passive or active allergic sensitization and challenge.