Showing papers by "National Jewish Health published in 2001"

A 12-week study of tacrolimus ointment for the treatment of atopic dermatitis in pediatric patients.

Abstract: The safety and efficacy of 0.03% and 0.1% tacrolimus ointment for the treatment of atopic dermatitis were evaluated in a 12-week, randomized, double-blind, vehicle-controlled study of 351 children 2 to 15 years of age with moderate to severe atopic dermatitis. The mean age of patients was 6.1 years. A total of 61.5% of patients had severe atopic dermatitis at baseline. The mean percentage of body surface area affected was 47.7%, and 83.5% of patients were affected on the head and/or neck. Significantly more patients (P<.001) achieved clinical improvement of 90% or better with 0.03% or 0.1% tacrolimus ointment compared with vehicle. Significant improvements in the signs and symptoms of atopic dermatitis, percent body surface area affected, and the patient's assessment of pruritus were also observed early in treatment and were maintained throughout the study. Adverse events with a statistically significantly greater incidence in the 0.03% tacrolimus ointment treatment group compared with vehicle were limited to the sensation of skin burning, pruritus, varicella, and vesiculobullous rash ("blisters"). Varicella and vesiculobullous rash occurred at a low incidence (<5%). No adverse event occurred at a statistically higher incidence in the 0.1% tacrolimus ointment-treated group compared with vehicle. Tacrolimus ointment was equally safe for younger (2-6 years) and older (7-15 years) children. Both tacrolimus ointment concentrations (0.03% and 0.1%) were safe and significantly more effective than vehicle for the treatment of atopic dermatitis in children.

A link between chronic asthma and chronic infection

Abstract: Background: Asthma is a prevalent disease with marked effects on quality of life and economic societal burden. However, the cause of asthma and its pathophysiology are not completely defined. Recently, the possibility that chronic infection may play a role has been suggested. Objective: We sought to define the association between Mycoplasma and Chlamydia species and chronic asthma. Methods: We performed a comparison study of asthmatic patients and normal control subjects. Fifty-five patients with chronic stable asthma were compared with 11 normal control subjects by using PCR, culture, and serology for Mycoplasma species, Chlamydia species, and viruses from the nasopharynx, lung, and blood. Bronchoalveolar lavage cell count and differential, as well as tissue morphometry, were also evaluated. Computer-generated scoring for the degree of chronic sinusitis in asthmatic patients was additionally evaluated. Results: Thirty-one of 55 asthmatic patients had positive PCR results for Mycoplasma (n = 25) or Chlamydia species (n = 6), which were mainly found on lung biopsy specimens or in lavage fluid. Only 1 of 11 normal control subjects had positive PCR results for Mycoplasma species. The distinguishing phenotype between asthmatic patients with positive and negative PCR results was the significantly greater number of tissue mast cells in the group with positive results. Conclusion: A significant number of patients with chronic stable asthma demonstrate the presence of Mycoplasma species, Chlamydia species, or both in their airways, with the distinguishing feature of increased mast cell number. These findings need further delineation but may help us to understand the pathophysiology of asthma and new treatment options. (J Allergy Clin Immunol 2001;107:595-601.)

Temporal association between airway hyperresponsiveness and airway eosinophilia in ovalbumin-sensitized mice.

Abstract: The temporal association between airway inflammation and airway hyperresponsiveness (AHR) has been analyzed in BALB/c mice sensitized to, and subsequently exposed to, a single intranasal challenge of ovalbumin (OVA). In OVA-sensitized/challenged animals only a small increase in responsiveness to methacholine (MCh) was seen at 8 h, peaked at 24 to 48 h, and resolved by 96 h. An early bronchoalveolar lavage fluid (BALF) neutrophil infiltrate (peaking at 8 h postchallenge; approximately 72% total cells was observed) that returned to baseline by 48 h. BALF eosinophil numbers did not increase until 48 h (approximately 32% of total cells), peaked at 96 h (approximately 38% total cells), and remained elevated at 8 d (approximately 27% total cells). Airway tissue eosinophilia preceded changes in BALF. Eosinophil peroxidase (EPO) levels in BALF were elevated in OVA-sensitized/challenged mice at 48 h only. BALF TNF-alpha levels peaked at 8 h, whereas IL-5 and IL-4 levels peaked at 24 h. IL-13 levels were increased at both 24 and 48 h. Mucus-positive cells were not observed in the airway epithelium until 48 h. Administration of IL-5 or VLA-4 antibody prior to OVA challenge prevented the development of AHR in sensitized mice as well as BALF and tissue eosinophilia. These data identify a temporal association between Th2 cytokine production, tissue eosinophil infiltration and activation, and, importantly, both the development and resolution kinetics of AHR. Moreover, the antibody studies further support the association of eosinophilia with the pathogenesis of AHR.

Formation of supramolecular activation clusters on fresh ex vivo CD8+ T cells after engagement of the T cell antigen receptor and CD8 by antigen-presenting cells.

Abstract: Upon productive interaction of CD4 T cells with antigen-presenting cells (APCs), receptors and intracellular proteins translocate and form spatially segregated supramolecular activation clusters (SMACs). It is not known whether SMACs are required for CD8 T cell activation. CD8 T cells, unlike CD4 T cells, can be activated by a single peptide-MHC molecule, or by purified monovalent recombinant peptide-MHC molecules. We studied, by three-dimensional digital microscopy, cell conjugates of fresh ex vivo CD8 T cells (obtained from OT-1 mice, which are transgenic for T cell antigen receptor reactive with the complex of H-2Kb and the ovalbumin octapeptide SIINFEKL) and peptide-pulsed APCs. Remarkably, even in T cell:APC conjugates that were formed in the presence of the lowest concentration of peptide that was sufficient to elicit T cell proliferation and IFN-γ production; the θ isoform of protein kinase C was clustered in a central SMAC, and lymphocyte function-associated antigen 1 and talin were clustered in the peripheral SMAC. Conjugation of T cells to APCs that were pulsed with concentrations of peptide smaller than that required to activate T cells was greatly reduced, and SMACs were not formed at all. APCs expressing mutant H-2Kb (Lys227) molecules that do not bind CD8 were unable to form stable conjugates with these T cells, even at high peptide concentrations. Thus, although CD8 and CD4 T cells may display different sensitivity to the concentration and oligomerization of surface receptors, SMACs are formed and seem to be required functionally in both cell types. However, unlike CD4 T cells, which can form SMACs without CD4, CD8 T cells from OT-1 transgenic mice depend on their coreceptor, CD8, for the proper formation of SMACs .

Inhibition of phosphodiesterase 4 attenuates airway hyperresponsiveness and airway inflammation in a model of secondary allergen challenge.

Abstract: We compared for the first time the therapeutic potential of a specific phosphodiesterase 4 (PDE4) inhibitor, rolipram, with anti-VLA-4 and anti-IL-5 in a model of secondary allergen exposure of previously sensitized and challenged mice. To address these issues, mice were sensitized and challenged with ovalbumin (OVA) (primary challenge). Six weeks later, sensitized/challenged mice were reexposed to OVA (secondary challenge) and airway response (resistance [RL] and dynamic compliance [Cdyn]) to inhaled methacholine was monitored. After secondary OVA challenge, RL significantly increased as did the number of lung inflammatory cells and IL-4 and IL-5 production in bronchoalveolar lavage fluid (BALF). Administration of rolipram, in a dose-dependent manner, significantly prevented both changes in RL and Cdyn, as well as eosinophil, lymphocyte, and neutrophil accumulation in the BALF; IL-4 and IL-5 levels in BALF were also significantly reduced. In contrast, treatment with anti-VLA-4 and anti-IL-5 only prevented changes in RL and eosinophil numbers and IL-5 production in BALF. Further, goblet cell hyperplasia was suppressed only by treatment with rolipram. None of the treatments affected OVA-specific antibody levels. These studies confirm that IL-5 dependent eosinophilic inflammation plays an essential role in the development of certain aspects of airway function after rechallenge of sensitized mice and that lymphocytes and neutrophils are also important in the development of altered airway function. The use of agents that inhibit PDE4 may have an important role in the treatment of asthma in previously sensitized mice.

Distal lung dysfunction at night in nocturnal asthma.

Abstract: We have previously shown that patients with nocturnal worsening of asthma (nocturnal asthma) exhibit increased parenchymal inflammation at night. To evaluate the functional significance of this parenchymal inflammation, 10 subjects with nocturnal asthma (NA), four subjects with non-nocturnal asthma (NNA), and four normal control subjects underwent bronchoscopy with measurement of peripheral airways resistance (Rp) at 4:00 p.m. and at 4:00 a.m. Employing a wedged bronchoscope technique, Rp was measured. Flow was stopped, and the pressure reached after 10 s of decay was termed the plateau pressure. The time constant of this decay ( τ ) was measured, and the peripheral compliance (Cp) was calculated as τ /Rp. The NA group exhibited the highest Rp values at 4:00 p.m. and at 4:00 a.m. as compared with the NNA and control groups, but all groups were significantly different from each other at 4:00 p.m.: NA, 0.113 ± 0.02 cm H2O/ml/min; NNA, 0.033 ± 0.005 cm H2O/ml/min; Control subjects, 0.010 ± 0.001 cm H2O/ ml/m...

Cardioprotective effect afforded by transient exposure to phosphodiesterase III inhibitors : the role of protein kinase A and p38 mitogen-activated protein kinase

Abstract: Background — Phosphodiesterase III inhibitors (PDEIII-Is) improve the hemodynamic status of heart failure via inotropic/vasodilatory effects attributable to the increase in intracellular cAMP level. Direct cardioprotection by PDEIII-Is and its underlying mechanisms, however, have not been identified. We tested the infarct size–limiting effect of PDEIII-Is and the roles of cAMP, protein kinase (PK) A, PKC, and mitogen-activated protein kinase (MAPK) families in open-chest dogs. Methods and Results — Milrinone, olprinone (PDEIII-Is), or dibutyryl-cAMP (db-cAMP) was injected intravenously 30 minutes before 90-minute ischemia, followed by 6 hours of reperfusion. Olprinone was also examined with an intracoronary cotreatment with a PKA inhibitor (H89), a PKC inhibitor (GF109203X), an extracellular signal–regulated kinase kinase (MEK) inhibitor (PD98059), or a p38 MAPK inhibitor (SB203580) throughout the preischemic period. Either PDEIII-Is or db-cAMP caused substantial hemodynamic changes, which returned to control levels in 30 minutes. Collateral flow and percent risk area were identical for all groups. Both PDEIII-Is and db-cAMP increased myocardial p38 MAPK activity during the preischemic period, which was blocked by H89, but not by GF109203X. Both PDEIII-Is and db-cAMP reduced infarct size (19.1±4.1%, 17.5±3.3%, and 20.3±4.8%, respectively, versus 36.1±6.2% control, P <0.05 each). Furthermore, the effect of olprinone was blunted by either H89 (35.5±6.4%) or SB203580 (32.6±5.9%), but not by GF109203X or PD98059. H89, GF109203X, PD98059, or SB203580 alone did not influence infarct size. Conclusions — Pretreatment with PDEIII-Is has cardioprotective effects via cAMP-, PKA-, and p38 MAPK–dependent but PKC-independent mechanisms in canine hearts. Received January 26, 2001; revision received April 10, 2001; accepted April 11, 2001.

Effect of Polymorphism of the β2-Adrenergic Receptor on Response to Regular Use of Albuterol in Asthma

Abstract: Background: Regular use of inhaled β-adrenergic agonists may have adverse effects in some asthma patients. Polymorphisms of the β2-adrenergic receptor (β2-AR)

Role of MEKK2-MEK5 in the regulation of TNF-α gene expression and MEKK2-MKK7 in the activation of c-Jun N-terminal kinase in mast cells

Abstract: Cross-linking of the high-affinity IgE receptor (FcɛRI) on mast cells with IgE and multivalent antigen triggers mitogen-activated protein (MAP) kinase activation and cytokine gene expression. We report here that MAP kinase kinase 4 (MKK4) gene disruption does not affect either MAP kinase activation or cytokine gene expression in response to cross-linking of FcɛRI in embryonic stem cell-derived mast cells. MKK7 is activated in response to cross-linking of FcɛRI, and this activation is inhibited by MAP/ERK kinase (MEK) kinase 2 (MEKK2) gene disruption. In addition, expression of kinase-inactive MKK7 in the murine mast cell line MC/9 inhibits c-Jun NH2-terminal kinase (JNK) activation in response to cross-linking of FcɛRI, whereas expression of kinase-inactive MKK4 does not affect JNK activation by this stimulus. However, FcɛRI-induced activation of the tumor necrosis factor-α (TNF-α) gene promoter is not affected by expression of kinase-inactive MKK7. We describe an alternative pathway by which MEKK2 activates MEK5 and big MAP kinase1/extracellular signal-regulated kinase 5 in addition to MKK7 and JNK, and interruption of this pathway inhibits TNF-α promoter activation. These findings suggest that JNK activation by antigen cross-linking is dependent on the MEKK2-MKK7 pathway, and cytokine production in mast cells is regulated in part by the signaling complex MEKK2-MEK5-ERK5.

Decreased steroid responsiveness at night in nocturnal asthma. Is the macrophage responsible

Abstract: As peripheral blood mononuclear cells from patients with nocturnal asthma (NA) exhibit reduced steroid responsiveness at 4:00 a.m. as compared with 4:00 p.m., we hypothesized that NA is associated with increased nocturnal airway cell expression of GR β , an endogenous inhibitor of steroid action. Ten subjects with NA and seven subjects with nonnocturnal asthma (NNA) underwent bronchoscopy with bronchoalveolar lavage (BAL) at 4:00 p.m. and 4:00 a.m. BAL lymphocytes and macrophages were incubated with dexamethasone (DEX) at 10− 5 to 10− 8 M. DEX suppressed proliferation of BAL lymphocytes similarly at 4:00 p.m. and 4:00 a.m. in both groups. However, BAL macrophages from NA exhibited less suppression of IL-8 and TNF- α production by DEX at 4:00 a.m. as compared with 4:00 p.m. (p = 0.0001), whereas in the NNA group DEX suppressed IL-8 and TNF- α production equally at both time points. GR β expression was increased at night only in NA, primarily due to significantly increased expression by BAL macrophages (p =...

ATOPIC DERMATITIS: New insights and opportunities for therapeutic intervention

Abstract: Atopic dermatitis (AD) is a chronic inflammatory skin disease that frequently predates the development of allergic rhinitis or asthma. It is an important skin condition with significant costs and morbidity to patients and their families; the disease affects more than 10% of children. Recent studies have demonstrated the complex interrelationship of genetic, environmental, skin barrier, pharmacologic, psychologic, and immunologic factors that contribute to the development and severity of AD. The current review will examine the cellular and molecular mechanisms that contribute to AD as well as the immunologic triggers involved in its pathogenesis. These insights provide new opportunities for therapeutic intervention in this common skin condition. (J Allergy Clin Immunol 2000;105:860-76.)

Juvenile arthritis and autoimmunity to type II collagen

Abstract: Objective Joint inflammation in juvenile rheumatoid arthritis (JRA) is sometimes associated with an autoimmune response to type II collagen (CII), a cartilage-specific protein. To test the hypothesis that down-regulation of autoimmunity to CII can be accomplished in JRA by oral administration of CII, an open-label study of CII was performed in 9 patients with JRA. Methods Seven rheumatoid factor–negative JRA patients with polyarticular disease and 2 JRA patients with pauciarticular disease (1 with early onset and 1 with late onset) were treated for 3 months with oral bovine CII. Patients were examined for disease activity and underwent routine laboratory testing at monthly intervals. Two of the patients had flares of disease when treatment was discontinued, and these patients were re-treated for an additional 3 months. To test the hypothesis that oral tolerance induces an immune deviation of T cells, peripheral blood mononuclear cells from patients were collected before and after treatment and cultured with CII. Supernatants and RNA were collected and analyzed for the presence of various cytokines. Results Eight patient trials met the criteria for clinical improvement outlined by Giannini and coworkers in 1997. None of the patients had any side effects from the treatment. In 6 of the 8 patients who improved, interferon-γ production decreased after oral CII therapy, correlating with clinical improvement, while 6 patients had increases in levels of transforming growth factor β3. Conclusion These results are encouraging. The possible beneficial effect of oral CII in JRA merits further investigation.

Method to inhibit airway hyperresponsiveness using aerosolized t cell receptor antibodies

Abstract: Disclosed is a method to reduce airway hyperresponsivesness in an animal by the direct delivery to the lungs of aerosolized antibodies against T cell receptors. The method is particularly useful for treating airway hyperresponsiveness associated with allergic inflammation, is effective at extremely low doses of antibody, and does not have a substantial effect on the peripheral immune system.

Tumor Necrosis Factor- α Negatively Regulates Airway Hyperresponsiveness through γδ T Cells

Abstract: Tumor necrosis factor (TNF)- α is a potent cytokine with immunomodulatory, proinflammatory, and pathobiologic activities. Although TNF- α is thought to play a role in mediating airway inflammation and airway hyperresponsiveness (AHR), its function is not well defined. TNF- α –deficient mice and mice expressing TNF- α in their lungs because of a TNF- α transgene placed under the control of the surfactant protein (SP)-C promoter (SP-C/TNF- α –transgenic mice) were sensitized to ovalbumin (OVA) and subsequently challenged with OVA via the airways; airway function in response to inhaled methacholine was monitored. In the TNF- α –deficient mice, AHR was significantly increased over that in controls. In contrast, the transgenic mice failed to develop AHR. In addition, sensitized/ challenged TNF- α –deficient mice had significantly increased numbers of eosinophils and higher levels of interleukin (IL)-5 and IL-10 in their bronchoalveolar lavage fluid than were found for control mice. However, in SP-C/TNF- α –tra...

Safety of aerosolized INS 365 in patients with mild to moderate cystic fibrosis: results of a phase I multi-center study.

Abstract: Cystic fibrosis (CF) is characterized by defective cystic fibrosis transmembrane regulator (CFTR) expression and function, associated with abnormal ion transport and mucociliary clearance, and clinical lung disease. Triphosphate nucleotides such as uridine-5′-triphosphate (UTP) and INS 365, may be useful for CF through actions, mediated via P2Y2 extracellular receptors, on chloride and liquid secretion, and ciliary beat frequency. INS 365 may offer chemical stability advantages over UTP. In a randomized, double-blind, multicenter phase I study, we studied the safety and maximally tolerated dose of escalating, single doses of aerosolized INS 365, in adult and pediatric patients with mild to moderate CF lung disease (FEV1 ≥ 45% predicted). In four successive dose cohorts of adult patients (n = 12 per cohort, age ≥ 18 years) and four successive pediatric dose cohorts (n = 12 per cohort, age 5–12 years), patients were randomized 3:1 active/placebo (0.9% saline) to evaluate doses of 20, 40, 80, and 100 mg INS 365 delivered by nebulizer (Pari Star™). Sputum was collected pre- and post-dosing to obtain preliminary results on clinical efficacy. After each dose cohort, a Data Safety Monitoring Committee (DSMC) reviewed the data. Forty-eight adult and 36 pediatric patients completed the protocol (up to 100 mg for adults, 80 mg for pediatric patients). The predominant adverse events were cough, wheezing, chest tightness, and a decrease in FEV1 (occurring in 8/48 adults, and 5/36 pediatric patients), which occurred predominantly in the 80-mg and 100-mg dose cohorts. Though a few adult patients had a tendency to increase sputum production, there was little consistent effect noted on sputum production in this acute, single-dose study. The data suggest that aerosolized INS 365 is safe when delivered at single doses of up to 40 mg in adults and children with CF, but that higher doses are unlikely to be tolerated. Pediatr Pulmonol. 2001; 32:122–128. © 2001 Wiley-Liss, Inc.

Method for identification of cell growth or differentiation factors

Abstract: The present invention relates to novel immortalized precursor cell populations derived from embryonic stem cell populations and methods to produce such cell populations. Also disclosed is an assay to identify regulatory compounds capable of controlling cell growth for therapeutic and experimental use.

γδ T cells as regulators of airway hyperresponsiveness

Abstract: Airway responsiveness (AR) is determined by complex mechanisms reflecting lung responses to airborne stimuli. Murine studies have identified a number of potential factors modulating AR and thus have contributed to the current understanding of these mechanisms. In allergic inflammation, immune cells, in particular αβ T cells, have emerged as important contributors to increased AR. We have found that in contrast to αβ T cells, γδ T cells can have a negative regulatory effect on AR. Here, we review the current studies on γδ T cells in allergic inflammation and discuss their role in modulating AR. We propose that γδ T cells exhibit different immune properties depending on the type of stimulus and inflammation. These differential immune properties appear to be associated with specific γδ T cell subsets, which control AR to airborne stimuli. In particular, our recent data indicate that the Vγ4 + T cell subset acts as an important negative regulator of AR and contributes to maintaining normal lung function in mice.

Phosphatidyl serine receptors and uses thereof

Abstract: Disclosed are phosphatidylserine (PS) receptors, including PS receptors from human, mouse, Drosophila melanogaster and Caenorhabditis elegans. Also disclosed are homologues of such receptors, nucleic acids encoding such receptors and homologues thereof, as well as agonist and antagonist compounds that specifically associate with and affect the activation state of such receptors. Preferred agonists and antagonists of PS receptors according to the invention include antibodies, antibody fragments and binding partners that selectively bind to such a receptor. Also disclosed are methods of making and using the PS receptors, homologues thereof, and agonist and antagonist compounds of such receptors. In particular, methods for reducing inflammation, for treating an autoimmune disease, for enhancing transplantation of tissue grafts, methods of increasing anti-tumor immunity, and methods for inhibiting viral and parasitic infections are described.

Method to limit immunodeficiency virus infection and compounds therefor

Abstract: Disclosed is a method to limit infection by an immunodeficiency virus. The method includes inhibiting an immunodeficiency virus protein which regulates apoptosis in cells. Also disclosed are methods to identify compounds that regulate cellular inhibitors of apoptosis in cells infected with an immunodeficiency virus and compounds identified thereby.

Long-Term Safety of Flunisolide Hydrofluoroalkane Metered-Dose Inhaler in Adults and Adolescents with Asthma

Abstract: To determine the long-term safety of flunisolide hydrofluoroalkane (HFA) in adults and adolescents with mild-to-moderate asthma. A 1-year multicentre, randomised, open-label, parallel-group study was conducted at 24 centres in the United States. A total of 215 male or female patients aged 12 to 60 years with mild-to-moderate asthma were eligible to enter the 52-week open-label phase. Eligible patients were randomly assigned in a 3:1 ratio to receive inhaled flunisolide (85μg per puff) or beclomethasone dipropionate chlorofluorocarbon (BDP CFC) [84μg per puff] as an active control. The incidence of adverse events was comparable in the flunisolide HFA (n = 162) and BDP (n = 53) groups; most were mild or moderate and considered unrelated to treatment. The flunisolide HFA group reported fewer respiratory complaints and had a significantly lower incidence of sinusitis (14.2 vs 26.4%, p = 0.04). No clinically important differences were observed for physical or laboratory parameters. The flunisolide HFA group had consistently fewer positive smears and cultures for Candida albicans, as well as a consistently lower incidence of candidiasis. Thrush infection occurred almost eight times more frequently in the BDP group (9.4 vs 1.2% for flunisolide HFA, p = 0.004). Flunisolide HFA, at one-third the daily dose (median daily dose of 340μg) of flunisolide CFC, was well tolerated and effective when administered to adult and adolescent patients with mild-to-moderate asthma.

Improved Methodology for Threshold Detection Studies in Asthmatic Children

Abstract: This paper reports the differences between two methodologies for threshold detection of added resistive loads in children and adolescents. The first-generation apparatus utilized a series of laminar flow screens to present various total resistances, while in the second generation the apparatus utilized a servo-controlled cone that occluded an aperture to varying degrees. Protocol modifications in the second generation methodology included forced choice, attentional enhancements, and larger increments of added resistance. Two studies conducted 2 years apart provided data on the first and second generations of methodology. All participants in Study 1 (N = 33) and Study 2 (N = 33) were children with asthma. Subjects were matched for both age and asthma severity. Results showed the methodologic improvements in the second generation to be significant. Tracking and random thresholds were achieved by 85% and 82% of the subjects in Study 2 compared to 76% and 42% in Study 1, respectively. The correlation...

The Measurement of Cysteinyl Leukotrienes in Urine

Abstract: The cysteinyl leukotrienes, comprising leukotriene (LT) C4 and its major metabolites LTD4 and LTE4, are inflammatory lipid mediators derived from metabolism of arachidonic acid by 5-lipoxygenase. These leukotrienes have received considerable attention for their potential role in asthma and other inflammatory diseases. Since there is a potential role for these lipid mediators in both health and disease, the analysis of leukotrienes in biological fluids, especially urine, has generated significant interest.