Induction of group IVC phospholipase A2 in allergic asthma: transcriptional regulation by TNFα in bronchoepithelial cells

摘要

pAirway inflammation in allergen-induced asthma is associated with eicosanoid release. These bioactive lipids exhibit anti- and pro-inflammatory activities with relevance to pulmonary pathophysiology. We hypothesized that sensitization/challenge using an extract from the ubiquitous fungus iAspergillus fumigatus/i in a mouse model of allergic asthma would result in altered phospholipase gene expression, thus modulating the downstream eicosanoid pathway. We observed the most significant induction in the group IVC PLAsub2/sub (phospholipase Asub2/sub) [also known as cPLAsub2/subγ (cytosolic PLAsub2/subγ) or PLA2G4C]. Our results infer that iA. fumigatus/i extract can induce cPLAsub2/subγ levels directly in eosinophils, whereas induction in lung epithelial cells is most likely to be a consequence of TNFα (tumour necrosis factor α) secretion by iA. fumigatus/i-activated macrophages. The mechanism of TNFα-dependent induction of cPLAsub2/subγ gene expression was elucidated through a combination of promoter deletions, ChIP (chromatin immunoprecipitation) and overexpression studies in human bronchoepithelial cells, leading to the identification of functionally relevant CRE (cAMP-response element), NF-κB (nuclear factor κB) and E-box promoter elements. ChIP analysis demonstrated that RNA polymerase II, ATF-2 (activating transcription factor 2)–c-Jun, p65–p65 and USF (upstream stimulating factor) 1–USF2 complexes are recruited to the cPLAsub2/subγ enhancer/promoter in response to TNFα, with overexpression and dominant-negative studies implying a strong level of co-operation and interplay between these factors. Overall, our results link cytokine-mediated alterations in cPLAsub2/subγ gene expression with allergic asthma and outline a complex regulatory mechanism./p