Characterization of Platelet-activating Factor Acetylhydrolase in Human Bronchoalveolar Lavage

Abstract

Platelet-activating factor (PAF) is a mediator produced in human airways during acute and chronic inflammatory lung diseases. The levels of PAF are regulated by acetylhydrolase (AH), the enzyme that converts PAF to lyso-PAF. To determine whether AH was present in human bronchoalveolar lavage (BAL) fluid, BAL was obtained from normal donors (n = 18) and from adult patients with mild bronchial asthma (n = 15) or with lung fibrosis (n = 15). AH activity was consistently found in the cell-free BAL fluid. BAL-AH is an enzyme different from secretory phospholipase A2 and from plasma AH and erythrocyte AH. Furthermore, BAL-AH is inhibited as much as 95% by exposure to an oxygen radical-generating system (xanthine/xanthine oxidase). BAL-AH is significantly correlated with the number of BAL macrophages (rs = 0.63; p < 0.02). In addition, BAL macrophages release AH both spontaneously and after stimulation with tumor necrosis factor-alpha (TNF-alpha) (100 ng/ml). BAL-AH activity in patients with bronchial asthma (1.32 +/- 0.18 pmol of PAF converted to lyso-PAF/min) is significantly lower than that in normal donors (2.25 +/- 0.26 pmol/min; p < 0.001). In contrast, BAL-AH activity in patients with lung fibrosis (6.13 +/- 0.81 pmol/min) is higher than that found in normal donors (p < 0.01). The variations in BAL-AH activity in patients with bronchial asthma or lung fibrosis are due to a reduction and to an increase, respectively, in the number of active molecules rather than to changes in enzyme affinity. These data demonstrate that human BAL fluid contains an extracellular AH activity that inactivates PAF released in the airways. BAL-AH is secreted by alveolar macrophages and is highly sensitive to oxygen radical-induced damage. The secretion and inactivation of BAL-AH may influence the levels of this enzyme in BAL fluid during acute and chronic inflammatory lung diseases and, ultimately, regulate the proinflammatory activities of PAF in these disorders.

Multiple Roles of Phospholipase A2during Lung Infection and Inflammation

“…Plasma PAF-AH is produced by cells of hematopoetic origin, predominantly macrophages, and its expression can be downregulated by LPS and certain cytokines (3,118). A deficiency of PAF-AH in the BALF of asthmatic patients compared to the level in healthy patients may prolong PAF activity and represent an underlying mechanism contributing to the asthmatic pathology (120). A point mutation in the plasma PAF-AH gene (V279F) results in an inactive enzyme and has been identified in patients from Japan.…”

Secretory Phospholipases A2 Induce β-Glucuronidase Release and IL-6 Production from Human Lung Macrophages

“…In selected experiments we used macrophages isolated from the bronchoalveolar lavage of patients undergoing diagnostic bronchoscopy for lung carcinoma. The bronchoalveolar lavage procedure and the isolation of macrophages were previously described (29). The cells were then incubated in 24-well plates (Falcon, Becton Dickinson, Franklin Lakes, NJ) at 37°C in a humidified atmosphere of 5% CO 2 and 95% air.…”

Association of PLA2G7 polymorphisms with carotid atherosclerosis in hypertensive Japanese

“…5 Plasma PAF-AH (pPAF-AH) has a relatively broad substrate specificity for phospholipids, and can catalyze and inactivate not only PAF but also PAF-like oxidized lipids. 6 This enzyme activity has been reported to correlate with several inflammatory diseases such as asthma, 7,8 systemic lupus erythematosus 9 and juvenile rheumatoid arthritis. 10 Furthermore, previous reports have revealed that the polymorphism of pPAF-AH gene (PLA2G7) influences its activity.…”