Nelly Frossard

Neurogenic mechanisms in bronchial inflammatory diseases

Neurogenic inflammation encompasses the release of neuropeptides from airway nerves leading to inflammatory effects. This neurogenic inflammatory response of the airways can be initiated by exogenous irritants such as cigarette smoke or gases and is characterized by a bi‐directional linkage between airway nerves and airway inflammation. The event of neurogenic inflammation may participate in the development and progression of chronic inflammatory airway diseases such as allergic asthma or chronic obstructive pulmonary disease (COPD). The molecular mechanisms underlying neurogenic inflammation are orchestrated by a large number of neuropeptides including tachykinins such as substance P and neurokinin A, or calcitonin gene‐related peptide. Also, other biologically active peptides such as neuropeptide tyrosine, vasoactive intestinal polypeptide or endogenous opioids may modulate the inflammatory response and recently, novel tachykinins such as virokinin and hemokinins were identified. Whereas the different aspects of neurogenic inflammation have been studied in detail in laboratory animal models, only little is known about the role of airway neurogenic inflammation in human diseases. However, different functional properties of airway nerves may be used as targets for future therapeutic strategies and recent clinical data indicates that novel dual receptor antagonists may be relevant new drugs for bronchial asthma or COPD.

Local increase in the number of mast cells and expression of nerve growth factor in the bronchus of asthmatic patients after repeated inhalation of allergen at low-dose.

Background Repeated inhalation of allergen at low‐dose induces an increase in bronchial hyper‐responsiveness, without any associated symptom. The concomitant events in the bronchus have not been described.

Thymic stromal lymphopoietin overproduced by keratinocytes in mouse skin aggravates experimental asthma.

Atopic dermatitis (AD) is often the initial step in the “atopic march,” given that more than half of AD patients with moderate to severe AD develop asthma later in life. Both AD and asthma share a similar “atopy” phenotype that includes T helper type 2 inflammation with eosinophilia and hyper-IgE immunoglobulinemia, but the molecular mechanisms underlying the “atopic march” remain elusive. In the present study, we show that induced expression of thymic stromal lymphopoietin (TSLP) in mouse epidermal keratinocytes upon topical application of MC903 (a low calcemic analogue of vitamin D3) not only triggers AD as we previously reported but also aggravates experimental allergic asthma induced by ovalbumin sensitization and challenge. Our study, which provides a mouse model to study human “atopic march,” indicates that keratinocyte-produced TSLP may represent an important factor in the link of atopic dermatitis to asthma.

Transforming growth factor-β and its role in asthma

Abstract Transforming growth factor-β (TGF-β) is an important fibrogenic and immunomodulatory factor that may play a role in the structural changes observed in the asthmatic airways. In vitro as well as in vivo studies have evidenced a dual role for TGF-β: it can either function as a pro- or anti-inflammatory cytokine on inflammatory cells, participating into the initiation and resultion of inflammatory and immune responses in the airways. TGF-β is also involved in the remodelling of the airway wall, and has in particular been related to the subepithelial fibrosis. TGF-β is produced in the airways by inflammatory cells infiltrated in the bronchial mucosa, as well as by structural cells of the airway wall including fibroblasts, epithelial, endothelial and smooth muscle cells. By releasing TGF-β, these different cell types may then participate into the increased levels of TGF-β observed in bronchoalveolar lavage fluid from asthmatic patients. Taken together, these results suggest that TGF-β may play a role in inflammation in asthma. However, as its role is dual in the modulation of inflammation, further studies are needed to elucidate the precise role of TGF-β in the airways.

Human lung fibroblasts secrete nerve growth factor: effect of inflammatory cytokines and glucocorticoids

Nerve growth factor (NGF) has recently been suggested to contribute to inflammation and bronchial hyperresponsiveness in asthma. However, the cell types capable of NGF production in the human lung and airways, as well as the regulatory role of pro-inflammatory cytokines and of glucocorticoids on NGF secretion in pulmonary cells, have not been described. Human pulmonary fibroblasts were cultured in the presence or absence of either interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and/or glucocorticoids. NGF secretion was measured by enzyme-linked immunosorbent assay. The human pulmonary fibroblasts constitutively secreted NGF in vitro. The rate of NGF secretion was shown to be cell density-dependent, since higher NGF secretion was detected in preconfluent cells, i.e. ones with less established cell-to-cell contact (41.0+/-5.0 pg x 10(-6) cells at 80% confluence), than cells in higher densities (8.2 +/- 3.4 pg x 10(-6) cells at 100% confluence). Stimulation with the pro-inflammatory cytokines IL-1beta (0.3-30 U x mL(-1)) or TNF-alpha (0.1-30 ng x mL(-1)) dose- and time-dependently (8-72 h) elevated the NGF secretion (effective concentration causing 50% of the maximum response (EC50)=2.9 U x mL(-1) and 1.0 ng x mL(-1), respectively). Treatment with the glucocorticoid budesonide (10(-7) M) markedly reduced the constitutive secretion of NGF by 42%, and attenuated the cytokine-stimulated NGF secretion to the same level. In conclusion, human lung fibroblasts may serve as a source of nerve growth factor in the lung, positively regulated by the asthma-associated and pro-inflammatory cytokines interleukin-1beta and tumour necrosis factor-alpha, and negatively regulated by the anti-inflammatory glucocorticoids.

Nerve growth factor levels and localisation in human asthmatic bronchi.

Nerve growth factor (NGF) has recently been suggested to be an important mediator of inflammation. In support of this, serum levels of NGF have been shown to be enhanced in asthmatics. However, it has not yet been shown whether the levels of NGF are also altered locally in asthmatic airways, when compared with healthy subjects, and the localisation of potential sources of NGF in the human bronchus have not yet been described. The aim of the present study was to assess NGF levels in bronchoalveolar lavage fluid (BALF) from asthmatics and to compare them to those of control subjects. Furthermore, the authors wanted to localise potential sources of NGF in bronchial tissue, and to number NGF-immunopositive infiltrating cells in the bronchial submucosa. BALF and bronchial biopsies were obtained from seven control subjects and seven asthmatic patients by fibreoptic bronchoscopy. NGF protein levels were quantified by enzyme-linked immunosorbent assay in BALF. NGF localisation was examined by immunohistochemistry on bronchial biopsy sections. The asthmatics exhibited significantly enhanced NGF levels in BALF. Intense NGF-immunoreactivity was observed in bronchial epithelium, smooth muscle cells and infiltrating inflammatory cells in the submucosa, and to a lesser extent in the connective tissue. The asthmatics exhibited a higher number of NGF-immunoreactive infiltrating cells in the bronchial submucosa than control subjects. This study provides evidence that nerve growth factor is locally produced in the airways, and shows that this production is enhanced in asthmatics. These findings suggest that nerve growth factor is produced by both structural cells and infiltrating inflammatory cells in human bronchus in vivo, and the authors suggest that the increase in nerve growth factor protein in bronchoalveolar lavage fluid observed in asthmatic patients may originate both from structural cells, producing increased nerve growth factor levels in inflammatory conditons, and from the increase in nerve growth factor-immunopositive cells determined in the bronchial submucosa.

Ser276 phosphorylation of NF-kB p65 by MSK1 controls SCF expression in inflammation.

Transcription of the mast cell growth factor SCF (stem cell factor) is upregulated in inflammatory conditions, and this is dependent upon NF-κB, as well as the MAP kinases p38 and ERK activation. We show here that the MAPK downstream nuclear kinase MSK1 induces NF-κB p65 Ser276 phosphorylation upon IL-1ß treatment, which was inhibited in cells transfected with a MSK1 kinase-dead (KD) mutant compared to the WT control. In addition, we show by ChIP experiments that MSK1 as well as MAPK inhibition abolishes binding of p65, of its coactivator CBP, and of MSK1 itself to the κB intronic enhancer site of the SCF gene. We show that interaction between NF-κB and CBP is prevented in cells transfected by a p65 S276C mutant. Finally, we demonstrate that both transfections of MSK1-KD and MSK1 siRNA - but not the WT MSK1 or control siRNA - downregulate the expression of SCF induced by IL-1ß. Our study provides therefore a direct link between MSK1-mediated phosphorylation of Ser276 p65 of NF-κB, allowing its binding to the SCF intronic enhancer, and pathophysiological SCF expression in inflammation.

Tachykinin receptors and the airways

The tachykinins, substance P, neurokinin A and neurokinin B, belong to a structural family of peptides. In mammalian airways, substance P and neurokinin A are colocalized to afferent C-fibres. Substance P-containing fibres are close to bronchial epithelium, smooth muscle, mucus glands and blood vessels. Sensory neuropeptides may be released locally, possibly as a result of a local reflex, and produce bronchial obstruction through activation of specific receptors on these various tissues. Three types of tachykinin receptors, namely NK-1, NK-2 and NK-3 receptors, have been characterized by preferential activation by substance P, neurokinin A and neurokinin B respectively. NK-1 and NK-2 receptors were recently cloned. The determination of receptor types involved in the effects of tachykinins in the airways has been done with synthetic agonists and antagonists binding specifically to NK-1, NK-2 and NK-3 receptors. Although the existence of species differences, the conclusion that bronchial smooth muscle contraction is mainly related to activation of NK-2 receptors on bronchial smooth muscle cell has been drawn. The hypothesis of a NK-2 receptor subclassification has been proposed with NK-2A receptor subtype in the guinea-pig airways. Other effects in the airways are related to stimulation of NK-1 receptors on mucus cells, vessels, epithelium and inflammatory cells. A non-receptor-mediated mechanism is also involved in the effect of substance P on inflammatory cells and mast cells.

Regulation of inflammation by PPARs: a future approach to treat lung inflammatory diseases?

Lung inflammatory diseases, such as acute lung injury (ALI), asthma, chronic obstructive pulmonary disease (COPD) and lung fibrosis, represent a major health problem worldwide. Although glucocorticoids are the most potent anti‐inflammatory drug in asthma, they exhibit major side effects and have poor activity in lung inflammatory disorders such as ALI or COPD. Therefore, there is growing need for the development of alternative or new therapies to treat inflammation in the lung. Peroxisome proliferator‐activated receptors (PPARs), including the three isotypes PPARα, PPARβ (or PPARδ) and PPARγ, are transcription factors belonging to the nuclear hormone receptor superfamily. PPARs, and in particular PPARα and PPARγ, are well known for their critical role in the regulation of energy homeostasis by controlling expression of a variety of genes involved in lipid and carbohydrate metabolism. Synthetic ligands of the two receptor isotypes, the fibrates and the thiazolidinediones, are clinically used to treat dyslipidaemia and type 2 diabetes, respectively. Recently however, PPARα and PPARγ have been shown to exert a potent anti‐inflammatory activity, mainly through their ability to downregulate pro‐inflammatory gene expression and inflammatory cell functions. The present article reviews the current knowledge of the role of PPARα and PPARγ in controlling inflammation, and presents different findings suggesting that PPARα and PPARγ activators may be helpful in the treatment of lung inflammatory diseases.

μ-opioid receptors modulate non-cholinergic constrictor nerves in guinea-pig airways

Abstract A role of opioids as inhibitors of the non-adrenergic non-cholinergic (NANC) excitatory nerves has been studied in the guinea-pig bronchi using electrical field stimulation. Morphine and DAGO gave dose-dependent inhibition of NANC contraction, which was reversed by naloxone, whereas the cholinergic nerve and substance P responses were unaffected. [Met5]enkephalin and [Leu5]enkephalin weakly inhibited NANC contraction, whereas dynorphin-(1–13) had no effect. These results suggest that opiates inhibit NANC bronchoconstriction via μ-opioid receptors.