Assays for histamine-releasing factors: from identification and cloning to discovery of binding partners.

When using a model to study disease, it may be advantageous to identify molecules responsible for biologic functions observed in the model to better understand the disease process being studied. The late phase reaction is used as a model for chronic inflammation, and the histamine releasing activity observed from late phase fluids was thought to be an important factor in the propagation of symptoms that remain in both the late-phase reaction and in chronic inflammation, when the offending antigen is no longer present. Purification from biologic fluids and identification may be helpful in understanding the role of the histamine-releasing factors in inflammation.

Synergistic interactions between airway afferent nerve subtypes regulating the cough reflex in guinea-pigs.

Cough initiated from the trachea and larynx in anaesthetized guinea-pigs is mediated by capsaicin-insensitive, mechanically sensitive vagal afferent neurones. Tachykinin-containing, capsaicin-sensitive C-fibres also innervate the airways and have been implicated in the cough reflex. Capsaicin-sensitive nerves act centrally and synergistically to modify reflex bronchospasm initiated by airway mechanoreceptor stimulation.

IgE and Fc{epsilon}RI regulation.

A central feature of allergic reactions is the aggregation of the high-affinity IgE receptor, FcepsilonRI, to initiate a change in the behavior of the cell expressing the receptor. It is now clear that a number of cell types can express this receptor, which broadens the biology that revolves around IgE antibody. It is also quite clear that the presence of monomeric IgE antibody alters the expression of FcepsilonRI.

Assessment of signal transducer and activator of transcription 6 as a target of glucocorticoid action in human airway epithelial cells.

Background: Activation of signal transducer and activator of transcription (STAT)6 by IL-4 and IL-13 is essential in many key epithelial responses in the asthmatic airway including expression of numerous chemokines, goblet cell differentiation and mucus production and expression of other allergic inflammatory genes. While these responses are all inhibited by glucocorticoids (GC) administered systemically or by inhalation, the inhibitory mechanisms are unknown.

Objective: To test the hypothesis that GC suppress allergic responses by blocking IL-4-induced STAT6 signalling in airway epithelial cells.

Capacitative calcium entry and TRPC channel proteins are expressed in rat distal pulmonary arterial smooth muscle.

Mammalian homologs of transient receptor potential (TRP) genes in Drosophila encode TRPC proteins, which make up cation channels that play several putative roles, including Ca2+ entry triggered by depletion of Ca2+ stores in endoplasmic reticulum (ER). This capacitative calcium entry (CCE) is thought to replenish Ca2+ stores and contribute to signaling in many tissues, including smooth muscle cells from main pulmonary artery (PASMCs); however, the roles of CCE and TRPC proteins in PASMCs from distal pulmonary arteries, which are thought to be the major site of pulmonary vasoreactivity, remain uncertain. As an initial test of the possibility that TRPC channels contribute to CCE and Ca2+ signaling in distal PASMCs, we measured [Ca2+]i by fura-2 fluorescence in primary cultures of myocytes isolated from rat intrapulmonary arteries (>4th generation).

IFN-alpha inhibits IL-3 priming of human basophil cytokine secretion but not leukotriene C4 and histamine release.

Background: Innate immune responses play a critical role in determining the course of acquired immunity, including that associated with allergic disease. Type I interferons, which are generated early in these reactions, are important soluble factors that prime for TH1-like activity.

Objective: Because human basophils secrete IL-4 and IL-13 in response to both IgE-dependent and IgE-independent stimuli, we tested whether IFN-alpha, a major type I IFN, affects the production of these TH2 cytokines and/or mediator release from these cells.

Inhibition of cytokine gene transcription by the human recombinant histamine-releasing factor in human T lymphocytes.

Human recombinant histamine-releasing factor (HrHRF) preincubation enhances the secretion of histamine, IL-4, and IL-13 from FcepsilonRI-stimulated human basophils. In GM-CSF-primed human eosinophils, HrHRF increases IL-8 production. Our recent experiments were designed to evaluate the effects of HrHRF on human T cell cytokine production.

Hypoxic constriction and reactive oxygen species in porcine distal pulmonary arteries.

To determine whether reactive oxygen species (ROS) play an essential role in hypoxic pulmonary vasoconstriction (HPV) and the cellular locus of ROS production and action during HPV, we measured internal diameter (ID) at constant transmural pressure, lucigenin-derived chemiluminescence (LDCL), and electron paramagnetic resonance (EPR) spin adduct spectra in small distal porcine pulmonary arteries, and dichlorofluorescein (DCF) fluorescence in myocytes isolated from these arteries. Hypoxia (4% O2) decreased ID, increased DCF fluorescence, tended to increase LDCL, and in some preparations produced EPR spectra consistent with hydroxyl and alkyl radicals. Superoxide dismutase (SOD, 150 U/ml) or SOD + catalase (CAT, 200 U/ml) did not alter ID during normoxia but reduced or abolished the constriction induced by hypoxia.

An in vivo guinea pig preparation for studying the autonomic regulation of airway smooth muscle tone.

The autonomic nervous system plays a primary role in regulating airway smooth muscle tone. Here, we describe the development of an in vivo guinea pig model that permits systematic studies of the autonomic control of airway smooth muscle. The model is based on preparations previously described and utilizes measurements of isometric tension in a perfused segment of extrathoracic guinea pig trachea in situ.

Association of the Src homology 2 domain-containing inositol 5' phosphatase (SHIP) to releasability in human basophils.

During the study of the biology of the Human recombinant Histamine Releasing Factor (HrHRF), we uncovered a hyperreleasable phenotype of basophils from HrHRF-responder donors. Basophils from these donors released histamne to HrHRF, IL-3 and D(2)O. While there has been a significant amount of work elucidating signal transduction events in human basophils, the reason for this hyperreleasable phenotype remained illusive.

Endogenous neurokinins facilitate synaptic transmission in guinea pig airway parasympathetic ganglia.

Neurokinin-containing nerve fibers were localized to guinea pig airway parasympathetic ganglia in control tissues but not in tissues pretreated with capsaicin. The purpose of the present study was to determine whether neurokinins, released during axonal reflexes or after antidromic afferent nerve stimulation, modulate ganglionic synaptic neurotransmission. The neurokinin type 3 (NK(3)) receptor antagonists SB-223412 and SR-142801 inhibited vagally mediated cholinergic contractions of bronchi in vitro at stimulation voltages threshold for preganglionic nerve activation but had no effect on vagally mediated contractions evoked at optimal voltage or field stimulation-induced contractions.

Central nervous system control of the airways: pharmacological implications.

Autonomic innervation of the airways is derived primarily from the parasympathetic nervous system. Preganglionic fibers originating in the brainstem project to parasympathetic ganglion neurons, which regulate airway smooth-muscle tone, glandular secretion and blood-vessel diameter. Airway preganglionic nerve activity is regulated by subsets of pulmonary and extrapulmonary afferent nerve fibers, which continuously provide polysynaptic input to brainstem preganglionic nuclei.

Evidence for differential reflex regulation of cholinergic and noncholinergic parasympathetic nerves innervating the airways.

The hypothesis that cholinergic and nonadrenergic, noncholinergic parasympathetic nerves innervating the airways are subject to differential reflex regulation was addressed. Pronounced contractile and relaxant parasympathetic reflex responses could be evoked by intravenous histamine, laryngeal mucosal application of capsaicin, inhaled capsaicin, or electrical stimulation of the vagal afferent nerves projecting to the esophagus and abdominal viscera. These data suggest that activation of multiple vagal afferent nerve subtypes can initiate both cholinergic and noncholinergic parasympathetic reflexes in the airways.

Expression of tachykinins in nonnociceptive vagal afferent neurons during respiratory viral infection in guinea pigs.

Immunohistochemistry was combined with retrograde labeling to characterize the effect of respiratory infection with Sendai virus on the number of Substance P/Neurokinin A-containing vagal afferent neurons whose cell bodies resided in the nodose ganglia and whose receptive fields were located in guinea pig trachea. Of the neurons labeled from the trachea of vehicle-inoculated guinea pigs, few stained positively for Substance P/Neurokinin A (approximately 3% of total labeled neurons). These neurons had small diameter cell bodies (mode = 16-20 microm), a feature of nociceptive-like C-fibers.

Allergic inflammation-induced neuropeptide production in rapidly adapting afferent nerves in guinea pig airways.

In the vagal-sensory system, neuropeptides such as substance P and calcitonin gene-related peptide (CGRP) are synthesized nearly exclusively in small-diameter nociceptive type C-fiber neurons. By definition, these neurons are designed to respond to noxious or tissue-damaging stimuli. A common feature of visceral inflammation is the elevation in production of sensory neuropeptides.

Nitric oxide-dependent modulation of smooth-muscle tone by airway parasympathetic nerves.

We addressed the hypothesis that noncholinergic parasympathetic nerves modulate airway smooth-muscle (ASM) tone in guinea pigs. The NO synthase inhibitor L-N(G)-nitro-arginine (L-NNA) and the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) potentiated cholinergic contractions and partly inhibited noncholinergic relaxations of the trachealis evoked by nerve stimulation in vitro or in situ. When delivered selectively to the trachea in situ, L-NNA and ODQ also increased baseline cholinergic tone of the trachealis, and L-NNA potentiated histamine-induced contractions of the trachealis in situ.

Histone deacetylation inhibits IL4 gene expression in T cells.

Background: Dysregulated expression of IL-4 has been linked with allergic diseases. IL-4 expression is controlled at the level of gene transcription by the coordinated action of multiple factors that bind regulatory promoter elements. In addition, alterations in chromatin structure are thought to play a role in regulating the expression of cytokines in the T(H)2 gene cluster, although the biochemical basis for these alterations in human T cells is not well understood.

Transient exposure of human bronchial epithelial cells to cytokines leads to persistent increased expression of ICAM-1.

Effects of several cytokines on kinetics of Intercellular Adhesion Molecule-1 (ICAM-1) and Vascular Cell Adhesion Molecule-1 (VCAM-1) expression were studied on a bronchial epithelial cell line (BEAS-2B). VCAM-I was neither constitutively expressed on BEAS-2B cells nor induced by Interferon-gamma (IFN-gamma). Tumor Necrosis Factor-alpha (TNF-alpha), Interleukin-1beta (IL-1beta), IFN-alpha, IL-4, IL-6, IL-8 or Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF).

Role of nerves in asthmatic inflammation and potential influence of gastroesophageal reflux disease.

Inflammation of the lower airways is a defining characteristic of asthma. Microaspiration of refluxate may initiate an inflammatory response in the airways of patients with gastroesophageal reflux disease (GERD), thereby precipitating asthma. Airway nerves are likely to play a role in the pathogenesis of asthma and could potentially mediate airway inflammation initiated by GERD through axonal reflexes.

Multiple mechanisms of reflex bronchospasm in guinea pigs.

The mechanisms of histamine- and bradykinin-induced reflex bronchospasm were determined in anesthetized guinea pigs. With intravenous administration, both autacoids evoked dose-dependent increases in tracheal cholinergic tone. Vagotomy or atropine prevented these tracheal reflexes.

Yin-Yang 1 activates interleukin-4 gene expression in T cells.

Interleukin-4 (IL-4) is a multifunctional cytokine that plays an important role in immune and inflammatory responses. Expression of the IL-4 gene is tightly controlled at the level of gene transcription by both positive and negative regulatory elements in the IL-4 promoter. Several constitutive nuclear factors have been identified that can interact with IL-4 promoter elements in DNA binding assays.

Inhibition of cytokine generation and mediator release by human basophils treated with desloratadine.

Background: Desloratadine is a non-sedating, clinically effective, anti-allergic therapy that has been shown to exhibit anti-inflammatory properties that extend beyond its ability to antagonize histamine at H(1)-receptor sites. This latter effect has been shown in vitro to be both IgE-dependent and -independent.

Objective: In this study, we addressed the ability of desloratadine to inhibit the in vitro generation of interleukin (IL)-4 and IL-13 from human basophils while concurrently comparing its efficacy in preventing mediator release by these cells.

Inflammation-induced plasticity of the afferent innervation of the airways.

The activation of primary afferent neurons that innervate the airways leads to homeostatic and defensive reflexes. The anatomic and physiologic characteristics of these afferent fibers do not appear to be static properties but rather appear to change rapidly in response to inflammation. The threshold for activation of airway afferent neurons to various stimuli, for example, is not fixed; these fibers can be become sensitized during inflammation.