concanavalin-a and Asthma

Nocturnal airway obstruction occurs frequently in childhood asthma and results from increased airway inflammation. Lymphocytes are believed to be key effector cells of airway wall inflammation, releasing pro-inflammatory mediators and cytokines. A previous study showed that hydrocortisone infusion, an effective anti-inflammatory treatment, improves nocturnal and daytime FEV(1) values. This study in 16 children with moderate asthma was designed to assess whether there exists day and night differences in IL-4, IL-5, IL-8, and IFN-gamma production of concanavaline A stimulated peripheral blood mononuclear cells. Furthermore, we investigated whether substitution of low serum cortisol levels with intravenous hydrocortisone would affect those parameters. Saline (as a placebo) or hydrocortisone (30 microg/m(2) body surface area/24 h) was intravenously administered in a randomized, double blind, cross-over design. Measurements under saline or hydrocortisone infusions were separated by 1 wk. At 04:00 and 16:00 hours 10 ml blood was taken for determination of peripheral blood mononuclear cell isolation and stimulation, and an eosinophil count. Hydrocortisone infusion significantly reduced the nocturnal fall in FEV(1). Median values of IFNgamma, IL-4, IL-5, and IL-8 produced by peripheral blood mononuclear cells did not significantly differ at 04:00 and 16:00 hours, both with saline and hydrocortisone infusion. Our results suggest that FEV(1) improvement is not due to suppression of circulating peripheral blood mononuclear cell activation. We hypothesize that it is rather due to its effect on local lung tissue epithelial and/or fibroblasts thereby reducing airway inflammation and vascular leakage.

Topics: Adolescent; Airway Obstruction; Anti-Inflammatory Agents; Asthma; Child; Circadian Rhythm; Concanavalin A; Cross-Over Studies; Cytokines; Double-Blind Method; Female; Forced Expiratory Volume; Humans; Hydrocortisone; Infusions, Intravenous; Interferon-gamma; Interleukin-4; Interleukin-5; Interleukin-8; Leukocytes, Mononuclear; Male; Time Factors

A group of 48 patients with asthma and rhinitis, sensitive to house dust allergen, underwent immunotherapy with an extract of crude house dust for a period of 6-12 months. The clinical results of the therapy, as evaluated by symptom medication scores, demonstrated the significant clinical improvement of the patients treated by crude dust extract (CDE) compared to those treated by placebo. A significant (p < 0.001) reduction of specific IgE and elevation (p < 0.001) of specific IgG in the post-therapeutic patient sera was demonstrated by serological tests. A good correlation was observed between the changes in specific IgE and IgG levels.

Topics: Adolescent; Adult; Asthma; Chromatography, Affinity; Chromatography, Gel; Concanavalin A; Desensitization, Immunologic; Dust; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin E; Immunoglobulin G; Injections, Subcutaneous; Male; Middle Aged; Rhinitis; Skin Tests

Metachromatic cells obtained from asthmatic subjects demonstrate increased spontaneous and stimulated histamine release in vitro. Their ability to synthesize and store proinflammatory cytokines has focused renewed interest on their role in asthma.. The late asthmatic response provides a useful model of clinical asthma. The aim of the study was to examine metachromatic cell derived mediators and histamine releasability in vitro after in vivo allergen exposure in atopic subjects with and without asthma and relate them to the type of physiological response observed.. Bronchoalveolar lavage (BAL) cells were obtained 4 h after challenge from asthmatics exhibiting a single early response (EAR, n = 5), a dual response (LAR, n = 7), unchallenged (basal, n = 5), atopic non-asthmatic (ANA, n = 6) and non-atopic non-asthmatics (normal, n = 5). BAL histamine and tryptase concentrations and in vitro histamine release (HR) after stimulation with anti-IgE, allergen, A23187, conconavalin A and substance P were compared.. Metachromatic cell numbers were lower in normal controls compared with all asthmatic groups and in LAR compared with EAR. Metachromatic cell derived mediators were higher in asthmatic compared with normal subjects. Spontaneous HR in LAR (20.5 +/- 5.0%) was lower than EAR (29.5 +/- 3.9%) and ANA (30.2 +/- 1.4%) (P < 0.05). No differences were seen in stimulated HR between EAR and LAR. HR in ANA stimulated with anti-IgE was greater than LAR (P < 0.05). HR in ANA stimulated with anti-IgE was greater than LAR (P < 0.05). After stimulation with ionophore A23187 (1 microM), release was greater in LAR compared with basal (P < 0.05) and no different at 5 microM. All subject groups responded to substance P (SP) but was significantly more in the asthmatic subjects compared to normal controls (P < 0.05). Allergen challenge did not modify the response of asthmatic subjects to SP.. Functional differences in metachromatic cell reactivity are present in atopic subjects 4h after in vivo allergen exposure which relate to the physiological response observed after this time and suggest that there is ongoing metachromatic cell degranulation subjects who subsequently develop LAR.

Topics: Adolescent; Adult; Allergens; Antibodies, Anti-Idiotypic; Asthma; Basophils; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Chymases; Concanavalin A; Female; Histamine Release; Humans; Inflammation Mediators; Leukocyte Count; Male; Mast Cells; Serine Endopeptidases; Substance P; Time Factors; Tryptases

A double-blind, randomized, crossover chronic study was done to determine the efficacy of colchicine in 10 atopic patients with asthma. A constant dose of sustained-release theophylline and albuterol by inhalation, as needed, was administered. Compared to placebo, colchicine, 0.5 mg twice daily, significantly reduced the mean (+/- SEM) daily clinical score from 2.18 +/- 0.34 to 1.64 +/- 0.32 (p less than 0.05), and the daily number of inhalations of albuterol from 5.89 +/- 1.48 to 4.01 +/- 1.26 (p less than 0.02). Colchicine significantly (p less than 0.05) increased the concanavalin A-induced suppressor cell function from 16.2 +/- 4.6% to 39.0 +/- 10.7%, which was similar to healthy volunteers (41.1 +/- 3.5%). Furthermore, colchicine significantly (p less than 0.05) decreased serum IgE from 248 +/- 63 to 188 +/- 46 IU/ml. Colchicine had no significant effect on pulmonary function tests, the early phase reaction of antigen-induced bronchial inhalation challenge, and immediate skin test responses. Thus, colchicine has immunomodulatory effects that may perhaps have a mild benefit in the treatment of asthma.

Topics: Adolescent; Adult; Albuterol; Asthma; Bronchial Provocation Tests; Chronic Disease; Colchicine; Concanavalin A; Delayed-Action Preparations; Double-Blind Method; Drug Therapy, Combination; Female; Humans; Lung; Male; Randomized Controlled Trials as Topic; Skin Tests; T-Lymphocytes, Regulatory; Theophylline

Fractional exhaled nitric oxide levels in exhaled breath can indicate ongoing eosinophilic airway inflammation, specifically in asthma. But its utility is being explored for central airway inflammations, including chronic obstructive pulmonary disease. Normal levels of fractional exhaled nitric oxide (F. F

Topics: Adult; Animals; Animals, Newborn; Asthma; Breath Tests; Choline; Concanavalin A; Demography; Diet; Exhalation; Female; Healthy Volunteers; Humans; Lactation; Lipopolysaccharides; Lymphocyte Count; Lymphocytes; Male; Mesentery; Middle Aged; Nepal; Nitric Oxide; Pulmonary Disease, Chronic Obstructive; Rats; Rats, Sprague-Dawley; Spleen; T-Lymphocytes

Enoxaparin, a low-molecular-weight heparin, is known to possess anti-inflammatory properties. However, its clinical exploitation as an anti-inflammatory agent is hampered by its anticoagulant effect and the associated risk of bleeding.. The aim of the current study was to examine the ability of non-anticoagulant fractions of enoxaparin to inhibit the release of key inflammatory cytokines in primed peripheral blood mononuclear cells derived from allergic mild asthmatics.. Peripheral blood mononuclear cells from allergic asthmatics were activated with phytohaemag glutinin (PHA), concanavalin-A (ConA) or phorbol 12-myristate 13-acetate (PMA) in the presence or absence of enoxaparin fractions before cytokine levels were quantified using specific cytokine bead arrays. Together with nuclear magnetic resonance analysis,time-dependent and target-specific effects of enoxaparin fractions were used to elucidate structural determinants for their anti-inflammatory effect and gain mechanistic insights into their anti-inflammatory activity.. Two non-anticoagulant fractions of enoxaparin were identified that significantly inhibited T-cell activation. A disaccharide fraction of enoxaparin inhibited the release of IL-4, IL-5, IL-13 and TNF-α by more than 57% while a tetrasaccharide fraction was found to inhibit the release of tested cytokines by more than 68%. Our data suggest that the observed response is likely to be due to an interaction of 6-O-sulfated tetrasaccharide with cellular receptor(s).. The two identified anti-inflammatory fractions lacked anticoagulant activity and are therefore not associated with risk of bleeding. The findings highlight the potential therapeutic use of enoxaparin-derived fractions, in particular tetrasaccharide, in patients with chronic inflammatory disorders.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Chemical Fractionation; Concanavalin A; Enoxaparin; Female; Humans; Interleukin-13; Interleukin-4; Interleukin-5; Leukocytes, Mononuclear; Male; Middle Aged; Oligosaccharides; Phytohemagglutinins; Primary Cell Culture; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha

Dietary fat intake has been associated with obesity and obesity in its turn with attenuated airway function and asthma, but it is unclear whether or how high-fat intake per se alters immune function relevant to development of allergic asthma.. To use a non-obese mouse model of mild to moderate allergic asthma to compare effects of high-fat with isocaloric control-diet on allergic immune responses.. C57BL/6 mice weaned and maintained on control (11% fat calories) or isocaloric high-fat diet (58% fat calories) were systemically sensitized with ovalbumin and challenged in the lungs. Allergic airway inflammation was assessed by measuring lung inflammation; serum antibodies; and, cytokines in serum, bronchoalveolar lavage (BAL) fluid and in supernatants of in vitro stimulated lung draining lymph node and spleen lymphocytes.. There was a significant reduction in lung eosinophilia and IL-5 in high-fat fed mice. Lung draining lymph node cells from these mice showed reduced pro-inflammatory cytokine (MCP-1 and TNF-alpha) release after ovalbumin re-stimulation and reduced release of IL-13 after concanavalin-A stimulation, indicating a general rather than just an antigen-specific change. There was no difference in IFN-gamma release. In contrast, pro-inflammatory cytokine release was increased from splenocytes. Decreased eosinophilia was not due to increased regulatory T cell or IL-10 induction in draining lymph nodes or spleen, nor to changes in antibody response to ovalbumin. However, decreased levels of serum and BAL eotaxin were found in high-fat fed animals.. The data indicate that high-fat dietary content redirects local immune responses to allergen in the lungs and systemic responses in the spleen and serum. These effects are not due to changes in regulatory T cell populations but may reflect a failure to mobilize eosinophils in response to allergic challenge.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Concanavalin A; Cytokines; Dietary Fats; Disease Models, Animal; Lung; Lymph Nodes; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Ovalbumin; Pneumonia; Pulmonary Eosinophilia; T-Lymphocytes, Regulatory

Previously we have reported that Y-27632, a selective inhibitor of Rho-associated protein kinases (ROCKs) for RhoA, suppressed concanavalin A (Con A)-induced secretion of cytokines from peripheral T cells of normal persons. Recent studies suggested its usefulness for clinical management of bronchial asthma. We examined effect of Y-27632 on release of cytokines from T cells of asthmatic patients. Peripheral T cells of six asthmatic subjects and six normal persons were stimulated with Con A for 24 h in the presence of Y-27632. The concentrations of IL-2, interferon (IFN)-gamma, IL-4, and IL-5 in supernatant were measured. Y-27632 down-regulated secretion of IL-2 and IFN-gamma and weakly decreased secretion of IL-4 and IL-5 from Con A-activated T cells in the asthmatics. The reduced secretion of IFN-gamma and IL-4 in the asthmatics was inferior to that in the normal subjects. Our data suggested that the involvement of RhoA/ROCK pathway in TCR-mediated secretion of IFN-gamma and IL-4 in patients with bronchial asthma differed from that in healthy persons.

Topics: Adult; Amides; Asthma; Cells, Cultured; Concanavalin A; Cytokines; Dose-Response Relationship, Drug; Female; Humans; Interferon-gamma; Interleukin-2; Interleukin-4; Interleukin-5; Intracellular Signaling Peptides and Proteins; Male; Protein Serine-Threonine Kinases; Pyridines; rho-Associated Kinases; T-Lymphocytes; Th1 Cells; Th2 Cells

The collectin surfactant protein D (SP-D) confers protection against pulmonary infection and inflammation. Recent data suggest a role for SP-D in the modulation of allergic inflammation.. The aim of this study is to characterize the immune responses of SP-D-deficient (SP-D(-/-)) mice in a kinetic model of allergic inflammation. We determined whether allergic parameters were enhanced in SP-D(-/-) mice in vivo. Further, we examined whether functional immune responses in vitro such as lymphocyte proliferation (LP) and cytokine production were modulated in the absence of SP-D.. In vivo, wild-type (WT) and SP-D(-/-) mice were sensitized and challenged with the allergen ovalbumin (OVA) and assessed for allergic parameters (bronchoalveolar lavage (BAL) eosinophils, IL-13 production, pulmonary IFN-gamma, IL-10 expression) at early time points (1 and 3 days of challenge) in comparison with late time points (7 days of challenge). In vitro, spleen cells from WT and SP-D(-/-) mice were stimulated with the mitogen concanavalin A (ConA) and lipid A (LpA) and analysed for LP, IL-13 and IFN-gamma production. Toll-like receptor 4 (TLR4), ligand for LpA, was assessed by mRNA expression and immunohistochemistry in vivo.. Following allergen exposure in vivo, SP-D(-/-) mice expressed higher BAL eosinophils and IL-13 concentrations and lower IFN-gamma expression at early time points compared with WT mice. IL-10 expression was increased at early time points in SP-D(-/-) compared with WT mice. Allergen-induced TLR4 expression was increased in WT, but not in SP-D(-/-) mice. After stimulation with LpA and ConA in vitro LP was increased and IFN-gamma concentration was decreased in SP-D(-/-) mice.. SP-D may be critical for the modulation of early stages of allergic inflammation in vivo.

Topics: Animals; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cell Proliferation; Concanavalin A; Eosinophils; Female; Immunoglobulin E; Interferon-gamma; Interleukin-10; Interleukin-13; Lipid A; Lymphocyte Activation; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Models, Animal; Ovalbumin; Pulmonary Surfactant-Associated Protein D; Receptors, Cell Surface; Toll-Like Receptor 4; Toll-Like Receptors

The regulatory mechanisms of IL-5 gene transcription in human peripheral T cells are unclear because the transfection efficiency of plasmid constructs into nontransformed T cells is very low.. Concanavalin A (ConA)-stimulated blastocytes derived from peripheral blood lymphocytes of asthmatic subjects were transiently transfected with the human IL-5 gene promoter/enhancer-luciferase gene construct, pIL-5 (-511)Luc, and cultured with THP-1 cells (human monocytoid cells) and anti-CD3 monoclonal antibody (mAb). IL-5 level in the culture medium was determined by an enzyme-linked immunosorbent assay. Transcriptional activity of the IL-5 gene was measured by luciferase reporter analysis.. ConA-blast lymphocytes of asthmatic patients produced a significant amount of IL-5 upon combined stimulation with anti-CD3 mAb and THP-1 cells, but not with anti-CD3 mAb alone. Costimulation with anti-CD28 mAb also enhanced the anti-CD3 mAb- induced IL-5 production. Accordingly, luciferase activity induced by anti-CD3 mAb stimulation in pIL-5(-511)Luc-transfected ConA-blast lymphocytes was increased 1.9- and 3.4-fold by the addition of anti-CD28 mAb and THP-1 cells, respectively. Serial 5' deletion analysis of the reporter gene demonstrated that the cis-regulatory element located at -119 to -80 is critical for anti-CD3 mAb-induced IL-5 gene transcription.. Our present findings provide a useful model reflecting IL-5 gene transcription in human peripheral T cells in vivo, and clearly demonstrate that an interaction with monocytes enhances IL-5 gene transcription.

Topics: Adult; Antibodies, Monoclonal; Asthma; Cell Adhesion Molecules; Concanavalin A; Down-Regulation; Gene Deletion; Gene Expression Regulation; Humans; Interleukin-5; Middle Aged; Monocytes; Promoter Regions, Genetic; Receptors, Antigen, T-Cell; Stimulation, Chemical; T-Lymphocytes; Transcription, Genetic

During the baseline period of a clinical trial comparing different dosage schedules of inhaled steroids, asthmatic children (aged 6-10 years) were expected to meet the inclusion criterion of airways hyper-responsiveness (PD(20) methacholine < 80 micro g) after withdrawal of inhaled corticosteroids for 2-8 weeks. However, many children failed to do so.. It has been shown that young wheezing children may outgrow their symptoms. We investigated if differences between children with and without airways hyper-responsiveness after withdrawal of inhaled corticosteroids were compatible with differences between transient and persistent wheezers found in other studies.. Seventy-eight children entered the study, of which 41 developed airways hyper- responsiveness after withdrawal of inhaled corticosteroids, and 37 did not. These two groups of children were compared with respect to differences in demographic, clinical, and immunological features (IL-4, IL-5, IL-10, and IFN-gamma produced by Con A stimulated peripheral mononuclear cells (PBMCs) and serum IL-4, IL-5 and soluble intercellular adhesion molecule-1 (sICAM-1)).. Hyper-responsive children had more atopic features (positive RAST, high IgE, eczema), lower levels of FEV1 and lower concentrations of sICAM-1 than non-hyper-responsive children. Apart from a borderline significantly higher IL-4 production in the hyper-responsive group, other immunologic parameters were comparable. Multivariate logistic regression analysis showed that high serum IgE, low FEV1, and low sICAM-1 levels were independently associated with the presence of airways hyper-responsiveness after stopping inhaled corticosteroids. Atopy was associated with higher concentrations of IL-4 in the hyper-responsive group.. After withdrawal of inhaled corticosteroids many children previously diagnosed with asthma did not develop airways hyper-responsiveness. We conclude that hyper-responsive children share features with persistent wheezers as found in previous studies, whereas the non-hyper- responsive children may represent transient wheezers.

Topics: Animals; Animals, Domestic; Asthma; Birth Weight; Breast Feeding; Breath Tests; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Cell Adhesion Molecules; Child; Concanavalin A; Female; Glucocorticoids; Humans; Hypersensitivity, Immediate; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-5; Leukocytes, Mononuclear; Logistic Models; Lymphocyte Activation; Male; Methacholine Chloride; Nitric Oxide; Respiratory Function Tests; Smoking; Unnecessary Procedures

The goal of our study was to examine spontaneous and stimulated apoptosis of peripheral blood MNC from allergic patients, sensitized to Der p I antigen as compared to cells from non-atopic subjects. Furthermore we aimed to investigate which populations of mononuclear cells (lymphocytes, monocytes) undergo the apoptosis and to determine relations between apoptosis and serum levels of sFas/APO-1, ICE/caspase-1 or TNF-alpha.. The study included 17 patients with perennial, allergic asthma and/or allergic rhinitis [6 male and 11 female; mean age 29,5 years; (range 15-49)]. Apoptosis was assessed by fluorescence technique and confirmed by flow-cytometric method and DNA ladder. Serum levels of sFas, ICE/caspase-1 or TNF-alpha were determined by immunoassays (ELISA).. Apoptotic index of unfractionated mononuclear cells (MNC) and lymphocytes (but not monocytes) were significantly higher in allergic patients as compared to non-allergic subjects after 48 and 72 hours of culture (p<0.05). Incubation of cells with ConA (10 microg/ml) resulted in a significant increase in the proportion of apoptotic cells in all populations once the apoptotic index for MNC and lymphocytes (but not monocytes) was again significantly higher in allergic as compared to non-allergic subjects after 24, 48 and 72 hour of culture. In allergic patients, mean serum sFas level, was significantly lower then in non-allergic group (mean value 624.8 pg/ml +/- 25.67 versus 802.0 pg/ml +/- 31.91; p = 0.003) and in both groups sFas level correlated inversely with apoptosis of MNC. The mean ICE/caspase-1 concentration was significantly higher in sera of allergic patients as compared to non-allergic group (mean value 27.71 pg/ml +/- 3.79 vs 23.54 pg/ml respectively; p<0.01). ICE/caspase-1 levels in allergic patients correlated with apoptotic index of mononuclear cells (r = 0.57; p<0.001).. An increased spontaneous and mitogen-induced apoptosis of MNC from peripheral blood of atopic patients as well as different serum levels of sFas and ICE/caspase-1 correlating with apoptosis, suggest different regulation of apoptotic process in peripheral blood mononuclear cells of patients with allergic asthma and/or rhinitis.

Topics: Adolescent; Adult; Apoptosis; Asthma; Caspase 1; Concanavalin A; fas Receptor; Female; Flow Cytometry; Humans; Leukocytes, Mononuclear; Male; Middle Aged; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha

Mechanisms that underlie the regulation of IL-5 gene expression in human peripheral T cells remain incompletely defined because of the low efficiency of transfection of plasmid constructs into non-transformed T cells. To elucidate the cellular and molecular mechanisms of IL-5 production, concanavalin A (ConA)-stimulated blastocytes derived from peripheral blood lymphocytes of asthmatic patients were employed in this study. Transcriptional activity of the synthetic human IL-5 promoter in ConA-stimulated blastocytes correlated with the production of IL-5. Deletion analysis of the reporter gene showed that the cis-regulatory element located at - 119 to - 80 is critical for inducible IL-5 promoter activity. Electrophoretic mobility shift assay revealed that an oligonucleotide probe corresponding to the element (- 119 to - 90) gave two specific bands. The slower migrating band was absolutely dependent on stimulation and was composed of a co-operative complex of the transcription factors, nuclear factor of activated T cells (NFAT) and activating protein-1 (AP-1). The faster migrating band was also inducible and was identified as AP-1-less NFAT. Mutation of either the NFAT or AP-1 element abrogated the slower migrating band and at the same time abolished transcriptional activity of the human IL-5 promoter/enhancer gene. Cyclosporin A equivalently suppressed DNA-binding activity of the composite NFAT/AP-1 site, promoter activity and protein production of IL-5. In conclusion, these data suggests that the composite NFAT/AP-1 binding element (- 115 to - 100) plays a crucial role in IL-5 synthesis by peripheral T cells of asthmatic patients.

Topics: Adult; Asthma; Cells, Cultured; Concanavalin A; DNA-Binding Proteins; Electrophoretic Mobility Shift Assay; Gene Expression Regulation; Humans; Interleukin-5; NFATC Transcription Factors; Nuclear Proteins; Promoter Regions, Genetic; T-Lymphocytes; Transcription Factor AP-1; Transcription Factors; Transcription, Genetic

IL-5 is strongly involved in the eosinophilic inflammation in bronchial asthma and atopic dermatitis. We have previously reported that IL-5 synthesis in atopic and nonatopic asthmatics is significantly enhanced compared to control subjects. T cell IL-5 synthesis is regulated through several transcriptional elements, one of which is the proximal human IL-5 promoter (-62 to -46). The present study was undertaken to delineate the transcriptional regulation through this element using nontransformed human T cells.. Con A blast lymphocytes which tolerate electroporation were derived from peripheral blood lymphocytes. Luciferase reporter analysis and gel shift analysis were performed.. The proximal promoter element is the overlapping binding site for the constitutive binding factor, Oct-1, and the inducible one, AP-1. The transcriptional induction was ascribed to the inducible binding, while the constitutive binding was rather inhibitory. A mutant element which lost the constitutive binding but retained the inducible binding exerted 3 times more transcriptional activity compared to the wild type element. In contrast, another mutant element which lost the inducible binding and retained the constitutive binding exhibited no transcriptional induction. Gel shift analysis clarified that the inducible binding was more prominent and the constitutive binding was less in IL-5-producing T cells derived from asthma patients compared to IL-5-nonproducing cells derived from control subjects.. The ratio of the inducible/constitutive binding to the proximal promoter element may determine the capacity of human Th cells to transcribe IL-5 gene, and its regulation may control eosinophilic inflammation.

Topics: Adult; Asthma; Base Sequence; Case-Control Studies; Concanavalin A; DNA; Humans; In Vitro Techniques; Interleukin-5; Lymphocyte Activation; Nucleic Acid Conformation; Promoter Regions, Genetic; T-Lymphocytes; Transcription, Genetic; Transfection

Asthma improves in most children during adolescence such that a small minority of patients exhibit clinically significant symptoms by the age of 20 years.. To investigate late allergic reactions, including eosinophil inflammation, associated with outgrowing mite antigen-induced bronchial asthma during adolescence, the relationship between clinical status and functional activity of interleukin (IL)-5 produced by Dermatophagoides farinae (Df)-stimulated peripheral mononuclear cells (PBMCs) in culture was assessed in mouse IL-3-dependent cells transfected with the human IL-5 receptor gene.. Activity of IL-5 spontaneously produced by PBMCs from either patients with mite-sensitive bronchial asthma or nonatopic control subjects was low. The activity of IL-5 produced by PBMCs stimulated with concanavalin A was significantly higher. Upon challenge with specific allergens, such as Df antigen, but not with irrelevant antigens, including ovalbumin, the in vitro activity was increased in patients with active disease and decreased in patients in remission.. Results suggest that the antigen-specific up-regulation of functional IL-5 activity in late allergic reactions is reduced in patients in remission and likely to result in an improvement in clinical status. The Df antigen may suppress Df-induced responses in patients with asthma in remission.

Topics: Adolescent; Adult; Animals; Antigens; Antigens, Dermatophagoides; Asthma; Cells, Cultured; Child; Concanavalin A; Epitopes; Female; Glycoproteins; Humans; Interleukin-5; Male; Mice; Mites; Monocytes

Intracellular levels of cAMP were found to regulate T cell activity. We examined whether beta2-agonists altered cytokine production and cyclic adenosine monophosphate (cAMP) accumulation in concanavalin A (ConA)-activated peripheral T cells from asthmatic patients. Procaterol and isoproterenol weakly decreased the ConA-elicited interleukin (IL)-4 and IL-5 secretion; however, the inhibitory effect of procaterol on the ConA-induced IL-2 secretion was inferior to that of isoproterenol in normal controls and was little in asthmatics. The intracellular accumulation of cAMP by procaterol was not altered compared with that by isoproterenol. Results suggest that there is a qualitative difference between procaterol- and isoproterenol-induced cAMP accumulation in T cells.

Topics: Adrenergic beta-Agonists; Asthma; Cells, Cultured; Concanavalin A; Cyclic AMP; Humans; Interleukin-2; Interleukin-4; Interleukin-5; Interleukins; Isoproterenol; Lymphocyte Activation; Procaterol; T-Lymphocytes

Immunity to Candida albicans (Candida) is characterized by a Th-1 type pattern of reactivity. Candida is rarely a cause antigen for bronchial asthma. Beta agonists have been found to inhibit secretion of IL-2 from T cells through intracellular cAMP elevation. We examined effects of isoproterenol (ISO) on Candida-stimulated T cells. Peripheral T cells obtained from six Candida-sensitive asthmatics, six Candida-sensitive non-asthmatic subjects, and six normal donors by Ficoll-Hypaque gradient centrifugation and nylon-wool column chromatography were incubated with Candida antigen or concanavalin A (Con A) in the absence or presence of ISO. Secretion of IL-2 and intracellular accumulation of cAMP were assayed by ELISA. Con A induced secretion of IL-2 in each of the three groups. Candida antigen induced IL-2 secretion in the normal and the non-asthmatic subjects, but not in the asthmatics. ISO, which reduced Con A-induced secretion of IL-2 in a dose-dependent manner, had no effect on Candida-induced secretion of IL-2. Although ISO increased the intracellular concentration of cAMP in untreated and Con A-treated T cells from all donors, cells from the normal and the non-asthmatic subjects, but not from the asthmatics, that were co-incubated with ISO and Candida had lower levels of cAMP than those treated with ISO alone. It is suggested that Candida antigen induces secretion of IL-2 and reduces ISO-inducible accumulation of cAMP in Candida-responsive IL-2 secreting cells, which may make Candida-induced secretion of IL-2 insensitive to ISO.

Topics: Asthma; Candida; Case-Control Studies; Concanavalin A; Cyclic AMP; Dose-Response Relationship, Drug; Humans; Hypersensitivity; In Vitro Techniques; Interleukin-2; Isoproterenol; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; T-Lymphocyte Subsets

Several anti-allergic anti-inflammatory drugs are used for the treatment of asthma including glucocorticosteroids (GCS), sodium cromoglycate (SCG), leukotriene (LT) inhibitors, and LT receptor antagonists. The major mechanism of the anti-inflammatory action of GCS is inhibition of cytokine production by T-lymphocytes: however, the mechanisms of anti-inflammatory effects of SCG are still unclear.. We elucidated the anti-inflammatory effects of SCG by investigating its effects on cytokine production by peripheral blood mononuclear cells (PBMCs) obtained from atopic asthmatics.. Peripheral blood mononuclear cells were obtained from seven atopic asthmatics and sensitized with Dermatophagoides farinae (DJ) or concanavalin A (ConA). We compared the effects of SCG on interleukin (IL)-5 and interferon (IFN)-gamma production by sensitized PBMCs with that of dexamethasone (Dex). Based on their clinical concentrations, we compared the effects of 10(-6) to 10(-4) M of SCG to those of 10(-6) M Dex.. Stimulation with ConA increased the production of IL-5 and IFN-gamma. Dex significantly inhibited the production of both cytokines but SCG showed no inhibitory effects. On the other hand, Df stimulation increased IL-5 production only. Dermatophagoides farinae-induced overproduction of IL-5 was inhibited by SCG and Dex.. Our results suggested that SCG has antigen-specific anti-allergic inflammatory effects.

Topics: Adult; Animals; Anti-Inflammatory Agents; Antigens; Antigens, Dermatophagoides; Asthma; Concanavalin A; Cromolyn Sodium; Cytokines; Dexamethasone; Female; Glycoproteins; Humans; Hypersensitivity, Immediate; Interferon-gamma; Interleukin-5; Leukocytes, Mononuclear; Male; Mites

In view of reports that CD8+ T cells may produce T(H2)-type cytokines and our own finding that levels of intracellular IL-4 are higher in CD8+ than CD4+ T cells in healthy nonatopic subjects, we have hypothesized that the capacity of CD8+ T cells to produce IL-4 may be increased in atopic asthma, a disease characterized by high production of T(H2) cytokines.. Levels of IL-4 and interferon-gamma were measured by ELISA in cell lysates and in 20- and 48-hour cultures of concanavalin A-stimulated purified peripheral blood CD8+ T cells in seven patients with mild atopic asthma and seven healthy nonatopic subjects.. Resting CD8+ T cells in patients with asthma contained significantly more IL-4 than those of healthy nonatopic subjects (median, 26 pg/10(6) cells; range, 17 to 84 pg/10(6) cells vs 16 pg/10(6) cells; 10 to 28 pg/10(6) cells), with no difference in intracellular interferon-gamma levels. In the healthy control subjects, but not in the patients with asthma, levels of intracellular IL-4 correlated negatively with levels of interferon-gamma in resting CD8+ T cells (r[s] = -0.9411, p = 0.005). Stimulation with concanavalin A produced a consistent and significant increase in secretion of interferon-gamma, but not IL-4, with no difference between the two groups of subjects.. The results of this study suggest that CD8+ T cells from patients with asthma may be an important source of the T(H2)-type cytokine IL-4. This capacity appears to be acquired in vivo, possibly by conditioning by IL-4 produced in the inflamed airways.

Topics: Asthma; CD8-Positive T-Lymphocytes; Cells, Cultured; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Humans; Interferon-gamma; Interleukin-4; Th2 Cells

The effect of T-440, a selective type IV phosphodiesterase inhibitor, on cytokine production by peripheral blood mononuclear cells (PBMC) of atopic asthmatics was investigated. T-440 suppressed allergen-induced interleukin (IL)-5 production with a high potency (IC50 = 0.039 microgram/ml) and allergen-induced proliferation of PBMC (IC50 = 0.30 microgram/ml). T-440 also suppressed IL-2, IL-4 and IL-5 production by concanavalin A-activated PBMC in concentration-dependent manner. The IC50 values for the suppression of cytokine synthesis were 0.11 microgram/ml for IL-2, 0.57 microgram/ml for IL-5 and 7.7 micrograms/ml for IL-4. cAMP-elevating agents, such as PGE2, forskolin and dibutyryl cAMP, suppressed IL-2, IL-4 and IL-5 production by concanavalin A-stimulated PBMC in a manner similar to that of T-440. T-440 inhibited cAMP-phosphodiesterase activity and raised the intracellular cAMP level of PBMC in a concentration-dependent manner, suggesting that the increase of intracellular cAMP caused by T-440 results in the reduction of cytokine production. We conclude that T-440 suppressed cytokine production by peripheral T lymphocytes via the protein kinase A pathway and may be an effective modality to treat atopic diseases associated with eosinophilic inflammation.

Topics: Adult; Asthma; Cells, Cultured; Concanavalin A; Cyclic AMP; Humans; Interleukin-2; Interleukin-4; Interleukin-5; Leukocytes, Mononuclear; Naphthalenes; Phosphodiesterase Inhibitors; Pyridones

Asthma and chronic bronchitis are associated with airway remodelling, and airway macrophages are present in bronchial inflammation. TGF-beta and fibronectin released by alveolar macrophages possess a fibrogenic potency. The potential role of alveolar macrophages in airway remodelling was studied in asthma and chronic bronchitis by the release of TGF-beta and fibronectin. Alveolar macrophages were isolated by bronchoalveolar lavage in 14 control subjects, 14 asthmatics and 14 chronic bronchitics. The spontaneous and lipopolysaccharide (LPS)- or concanavalin A (Con A)-induced release of TGF-beta and fibronectin was measured by ELISA. Alveolar macrophages from chronic bronchitics spontaneously release greater amounts of TGF-beta and fibronectin than those from asthmatic and control subjects. Alveolar macrophages from asthmatics release greater amounts of TGF-beta and fibronectin than those from control subjects. The spontaneous release of TGF-beta is significantly correlated with that of fibronectin. Fibronectin release was significantly reduced after LPS stimulation, and TGF-beta release was significantly increased after LPS stimulation, except in chronic bronchitis patients. Con A increased the release of TGF-beta in cells from normal subjects. This study suggests that activated macrophages play a role in airway remodelling in chronic bronchitis and to a lesser extent in asthma.

Topics: Adult; Asthma; Bronchitis; Chronic Disease; Concanavalin A; Female; Fibronectins; Humans; Lipopolysaccharides; Macrophages, Alveolar; Male; Middle Aged; Transforming Growth Factor beta

In atopic patients, allergen-sensitized T lymphocytes specifically proliferate in the presence of T cell-growth factor, interleukin 2 (IL-2). Lecithin-bound iodine (LBI), which has been used as a therapeutic modality for patients with bronchial asthma (BA), effectively inhibited Dermatophagoides farinae (Df) mite antigen-induced IL-2 responsiveness in concentrations comparable to LBI concentrations in blood. IL-2-responding T cells were more sensitive to LBI than antigen-presenting cells, whereas LBI suppressed the release of interleukin 1 (IL-1) elicited by Df antigen. In addition, ovalbumin (OVA)-induced IL-2 responsiveness in egg sensitive patients and purified protein-derivative (PPD)-induced IL-2 responsiveness were similarly inhibited by LBI. The IL-2 responsiveness induced by concanavalin A (Con A), however, was not changed. On the basis of these results, LBI may act as a slight immunosuppressant to inhibit the induction of allergen-specific lymphocytes and to improve the clinical status in allergic diseases.

Topics: Adult; Allergens; alpha-Linolenic Acid; Antigens, Dermatophagoides; Asthma; Child, Preschool; Concanavalin A; Dermatitis, Atopic; Epitopes; Glycoproteins; Humans; Hypersensitivity; Infant; Interleukin-1; Interleukin-2; Iodine; Lymphocyte Activation; Ovalbumin; Phosphatidylcholines; T-Lymphocytes; Tuberculin

Metachromatic cells are increased in the airways of asthmatic patients. We obtained metachromatic cells from asthmatic airways using an induced sputum technique. The histamine release following ConA, anti-IgE and anti-IgE + IL3 from those cells were evaluated before and following the addition of cromoline Na and nedocromil Na. Metachromatic cells had a higher rate of spontaneous histamine release when compared to peripheral basophils. Cromoline Na and nedocromil Na inhibited histamine release from triggered metachromatic cells but not from peripheral basophils. It is concluded that airway metachromatic cells from asthmatics behave differently than peripheral basophils.

Topics: Adult; Anti-Asthmatic Agents; Antibodies, Anti-Idiotypic; Asthma; Basophils; Bronchi; Concanavalin A; Cromolyn Sodium; Female; Forced Expiratory Volume; Histamine Release; Humans; Immunoglobulin E; Interleukin-3; Male; Nedocromil; Sputum

Enhanced activities of peripheral blood cells are a common characteristic of patients with asthma.. Here we tested whether this could be due to a dysfunction in one or more signal transduction systems.. The production of 1,2-diacylglycerol (1,2-DAG) and arachidonic acid was compared in mononuclear blood cells from patients with asthma (n = 10) and healthy controls (n = 12).. Using three different stimuli (concanavalin A, aluminium fluoride or the calcium ionophore A23187) no difference in the production of both 1,2-DAG and arachidonic acid could be found between patients and controls before allergen challenge. Concanavalin A-induced 1,2-DAG production could be inhibited completely in the presence of isoprenaline; concanavalin A-induced arachidonic acid production, partially. The inhibitory effect of adenylate cyclase activation on the production of 1,2-DAG and arachidonic acid was identical in patients and controls. Following allergen challenge, there was a tendency to an increased production of 1,2-DAG and arachidonic acid in controls, whereas in patients there was a tendency to a decreased production.. Enhanced cellular activities found in patients with asthma are not caused by an intrinsic dysfunction in production of 1,2-DAG and arachidonic acid.

Topics: Adult; Allergens; Animals; Arachidonic Acid; Asthma; Calcimycin; Concanavalin A; Diglycerides; Drug Interactions; Dust; Female; Fluorides; Humans; Isoproterenol; Leukocytes, Mononuclear; Male; Mites

The effects of IL-2, IL-3, and IL-4 activities on occurrence of allergic asthma induced by spores of mushroom have been studied by measuring the IL-2, IL-3, and IL-4 activities in culture supernatants of spleen lymphocytes from guinea pigs being stimulated with Con A. The IL-4 activity (14.9 +/- 0.18 u) in the culture supernatant was higher than control group (6.7 +/- 1.5 u). The IL-2 and IL-3 activities were all similar to that of the control group. The IL-4 synthesis was similarly raised as the IL-4 activity. These results indicated that there is a relationship between the occurrence of allergic asthma and the increase of IL-4.

Topics: Animals; Asthma; Cells, Cultured; Concanavalin A; Guinea Pigs; Hypersensitivity; Interleukin-2; Interleukin-3; Interleukin-4; Lymphocytes; Mice; Polyporaceae; Reference Values; Spleen; Spores, Fungal; Tumor Cells, Cultured

In order to identify and characterize the wheat grain allergens involved in bakers' asthma, proteins were sequentially extracted from whole-meal flour. The polypeptide composition of the individual solubility fractions (albumin/globulin, gliadin and glutenin) was analyzed by sodium dodecyl sulfate-gel electrophoresis (SDS-PAGE), and high-resolution two-dimensional gel electrophoresis with immobilized pH gradient 4-9 in the first dimension (IPG-Dalt). The resolved polypeptides were transferred onto an immobilizing polyvinylidene difluoride membrane and incubated with a pooled serum from four asthmatic bakers. Bound IgE was demonstrated by autoradiography using 125I-labeled anti-human IgE. Our study demonstrated that the serum of the bakers allergic to flour contained IgE antibodies which bound to numerous polypeptides of all three solubility fractions. The highest percentage of IgE binding was observed with certain albumin and/or globulin polypeptides, whereas the gliadins and glutenins exhibited considerably less allergenicity. SDS-PAGE revealed that the protein which bound the highest percentage of IgE from the sera of the allergic bakers is a 27 kDa albumin. More detailed investigations using IPG-Dalt demonstrated that this allergen is not a single polypeptide but consists of several polypeptide spots that differ in their isoelectric points. Quantitative studies using computer-assisted laser densitometry revealed that the amount of patients' IgE bound by these particular polypeptides differed considerably between the seven wheat cultivars examined, ranging from 13% to 53% of the total radioactive uptake.

Topics: Albumins; Allergens; Asthma; Concanavalin A; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Gliadin; Globulins; Glutens; Horseradish Peroxidase; Humans; Immunoenzyme Techniques; Immunoglobulin E; Occupational Diseases; Plant Lectins; Plant Proteins; Radioallergosorbent Test; Silver Staining; Triticum

Topics: Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis, Allergic Bronchopulmonary; Aspergillus fumigatus; Asthma; Concanavalin A; Humans; Immunoglobulin G

Occupational asthma due to Western Red Cedar (WRCA) is attributed to sensitization to plicatic acid (PA), but does not appear to be dependent on PA-specific IgE antibodies. Exposure to PA induces histamine release in vivo and in vitro, so if IgE is not important, other mechanisms of histamine release must presumably operate in WRCA. To explore the possible role of histamine-releasing factors in WRCA, peripheral blood mononuclear cells were obtained and cultured with PA, PA-albumin conjugate plicatic acid-human serum albumin (PA-HSA), grass pollen or Concanavalin A using a standard histamine releasing factor (HRF) generation protocol. Supernatants were dialysed to remove endogenous histamine and then assayed for histamine releasing activity using human basophils as targets and a Con A-induced bulk supernatant as an internal HRF standard. In contrast to some previous reports, spontaneous HRF release from the peripheral blood mononuclear cells (PBMC) of WRCA patients (n=9) and atopic asthmatic subjects (n=5) was not elevated compared with the non-asthmatic controls (n=11; five atopic and six non-atopic). Both PA and PA-HSA induced the production of small amounts of HRF by PBMC of WRCA patients, but a similar degree of HRF generation was also observed in PBMC from the atopic asthmatic, atopic nonasthmatic, and non-atopic subjects. In contrast, grass pollen induced the production of HRF by PBMC from the subjects with positive skin tests to grass pollen but not by PBMC of non-atopic subjects, confirming that our methods and assay were capable of detecting antigen-specific HRF production. Since neither PA nor PA-HSA induced significantly elevated HRF production from PBMC of WRCA patients, it seems unlikely that PA-induced HRFs play a substantial role in the pathogenesis of WRCA.

Topics: Adult; Allergens; Antibody Specificity; Asthma; Basophil Degranulation Test; Biological Factors; Bronchial Provocation Tests; Concanavalin A; Dust; Female; Histamine Release; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Leukocytes, Mononuclear; Lignans; Male; Middle Aged; Naphthols; Occupational Diseases; Plant Lectins; Poaceae; Pollen; Serum Albumin; Trees; Wood

This study had two purposes. First, to examine a possible functional heterogeneity of IgE regulating basophil histamine release and the effect of using two different donor cells for passive sensitization experiments. Second, to investigate basophils not releasing histamine to anti-IgE by stimulating protein kinase C with the addition of the phorbol-ester, TPA. In consecutive experiments responding donor basophils were passively sensitized with plasma from non-responding subjects. Thus, the first set of experiments included passive sensitization of acid treated donor basophils from one atopic and one non-atopic patient with plasma from 29 children with exogenous asthma to grass pollen, cat dander, or dust mites. Different secretagogues (anti-IgE, Concanavalin A, and N-formyl-methionyl-leucyl-phenylalanine) induced different histamine release responses due to a cellular property of the basophils not related to the type of IgE bound to the cell membrane. It was demonstrated that the allergen-induced histamine release did not depend on the extract or type of IgE when the biological activity of each extract and serum-specific IgE levels were similar. However, the atopic donor cells released significantly (P less than 0.05) more histamine than non-atopic donor cells. Thus, histamine release depends on the type of secretagogues and a cellular property which is maybe influenced by the presence of serum factors and a certain type of IgE in the serum of atopics. The second set of experiments included 10 patients (6 atopics and 4 non-atopics) with non-histamine releasing basophils. In the presence of 10 ng/ml TPA, however, seven of 10 patients released histamine at anti-IgE challenge. Three months later two additional patients became responsive in the presence of TPA. By passive sensitization of responding donor basophils the non-responding patients were shown to possess functionally intact IgE. Thus, the discrepancies sometimes observed between clinical symptoms, serological IgE-antibody measurements and histamine release testing in allergic patients may be related to a cellular property of basophils.

Topics: Adolescent; Asthma; Basophils; Child; Child, Preschool; Concanavalin A; Female; Histamine Release; Humans; Immunization, Passive; Immunoglobulin E; Male; N-Formylmethionine Leucyl-Phenylalanine

We tested the effect of Ketotifen (4-(1-methyl-4-piperidylidene)-4H- benzo[4,5] cyclohepta[1,2-b]thiophen-10(9H)-one hydrogen (fumarate) on the induction of allergen-induced IL-2 responsiveness in lymphocytes from patients with atopic dermatitis and/or bronchial asthma. Ovalbumin (OVA)- and/or Dermatophagoides farinae(Df)-induced IL-2 responsiveness was increased in almost all patients (1-15 years old) before Ketotifen treatment. Two to 12 months administration of Ketotifen (0.06 mg/kg/day) decreased activity of the response in 7 out of 9 cases corresponding to improvement of clinical symptoms. In in-vitro studies, antigen presenting cells (adherent cells) from the patient pretreated with 5, 50 and 500 ng/ml doses of Ketotifen for 12 h failed to present OVA or Df antigen to T-cells for induction of IL-2 responsiveness. Antigen-pulsed adherent cells also failed to induce the response of the T-cells pretreated with 50 and 500 ng/ml doses of Ketotifen but not with a 5 ng/ml dose. A 50 ng/ml dose of Ketotifen did not affect T-cells for induction of the response. In contrast, the treated adherent cells are capable of presenting PPD antigen or Con A for the induced response. The combined data indicate that induction of IL-2 responsiveness of peripheral blood lymphocytes on stimulation with nominal antigen may reflect an immune response to allergen in patients with allergy and a weak immunosuppressive effect of Ketotifen seems to block the response in the pathogenic process of allergic diseases.

Topics: Adolescent; Allergens; Asthma; Child; Child, Preschool; Concanavalin A; Dermatitis, Atopic; Female; Humans; Infant; Interleukin-2; Ketotifen; Male; Ovalbumin; T-Lymphocytes

Releasability of human basophils and mast cells is an important parameter in allergic disorders. We compared IgE- and non-IgE-mediated releasability of human peripheral blood basophils with that of mast cells obtained from lung parenchyma (isolated by mechanical or enzymatic dissociation) and from bronchoalveolar lavage of normal and asthmatic donors. In a first study, the response to anti-IgE, Staph A, Con A, f-met peptide, and Ca2+ ionophore A23187 of basophils obtained from 52 donors was compared with that of mast cells isolated enzymatically (PMCE) or mechanically (PMCM) from lung parenchyma obtained during surgery. The histamine content of basophils (1.1 +/- 0.1 pg/cell) was significantly lower than that of PMCE (4.1 +/- 0.3 pg/cell; p less than 0.001) and PMCM (3.7 +/- 0.3; p less than 0.001). The maximal percent anti-IgE-induced histamine secretion in basophils (41.3 +/- 3.6) was higher than in PMCE (17.5 +/- 1.8) and in PMCM (13.8 +/- 1.5). Similarly, the response to Staph A and Con A was higher in basophils (29 +/- 3.9 and 31.6 +/- 4.9, respectively) than in PMCE (3.5 +/- 0.6 and 3.3 +/- 0.8, respectively) and PMCM (5.1 +/- 1.3 and 8.8 +/- 2.2, respectively). A positive correlation between the maximal percent of histamine release induced by anti-IgE and Staph A was found in basophils (rs = 0.61; p less than 0.001), whereas there was a negative correlation between the reactivity of PMCE (rs = 0.67; p less than 0.001) and PMCM (rs = -0.40; p less than 0.001) to anti-IgE and their reactivity to Staph A.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Aged; Asthma; Basophils; Bronchoalveolar Lavage Fluid; Calcimycin; Cell Separation; Concanavalin A; Female; Histamine Release; Humans; Immunoglobulin E; Lung; Male; Mast Cells; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Staphylococcal Protein A

This study was undertaken to investigate the effect of ketotifen on the immune system of asthmatic children receiving prophylactic treatment with ketotifen. E. rosettes, mitogenic response to phytohemagglutinin (PHA) and concanavalin A (Con A) as well as Con A induced suppression were measured in 15 asthmatic patients before and 1-3 months after treatment with ketotifen (1 mg x 2/day), and in 20 normal healthy controls. No significant difference in immunological parameters studied were found between the pre and post treatment periods in the asthmatic patients, neither was any difference demonstrated between the asthmatic and control group studied.

Topics: Adolescent; Asthma; Child; Child, Preschool; Concanavalin A; Female; Humans; Immunity, Cellular; Ketotifen; Male; Phytohemagglutinins; Rosette Formation; T-Lymphocytes, Regulatory

Asthmatic patients have a deficiency of concanavalin A-(Con A) induced suppressor cell function. We tested whether oral colchicine 0.5 mg twice daily for 7 days could correct this immunoregulatory abnormality. Peripheral blood mononuclear cells were incubated with Con A and then suppression of proliferation was measured by coculture of these cells with healthy volunteers' mononuclear cells and phytohaemagglutinin. Sixteen asthmatic patients had significantly (P less than 0.002) decreased Con A-induced suppressor cell function (17.0 +/- 17.2%, mean +/- s.d.) as compared to 13 healthy volunteers (37.9 +/- 14.9%). Oral colchicine significantly (P less than 0.05) increased, though only partially corrected, these 16 asthmatic patients' Con A-induced suppressor cell function (28.1 +/- 14.3%). Asthmatic patients had an increased number of monocytes (691 +/- 289 vs 388 +/- 271/mm3 for normals, P less than 0.01) and a normal number of lymphocytes, Leu 4+ total T cells, Leu 3+ helper/inducer T cells, and Leu 2+ suppressor/cytotoxic T cells as well as a normal Leu 3/Leu 2 ratio. Oral colchicine significantly (P less than 0.005) decreased the number of monocytes (451 +/- 255/mm3) without significantly affecting the number of lymphocytes, Leu 4+, Leu 3+, or Leu 2+ T cells, or the Leu 3/Leu 2 ratio. These results are consistent with the hypothesis that the deficiency of Con A-induced suppressor cell function in asthmatic patients may be due, in part, to an increased number and/or abnormal activity of monocytes. If so, then oral colchicine may have partially corrected the deficiency of Con A-induced suppressor cell function by decreasing the number and/or modulating the activity of monocytes.

Topics: Adult; Asthma; Colchicine; Concanavalin A; Cyclic AMP; Female; Humans; Male; Middle Aged; Monocytes; T-Lymphocytes; T-Lymphocytes, Regulatory

IgE-dependent basophil histamine release does not necessarily correlate with the amount of cell-bound IgE, thus it has been suggested that basophil "releasability" is an important, but yet undefined, factor in this secretory process. Because mast cell, and possibly basophilic leukocyte, mediator release contributes to airway reactivity, any enhancement of this secretory process would favor asthma provocation. To evaluate IgE-dependent basophil histamine releasability in asthma, suspensions of leukocytes were isolated from patients with an allergic and nonallergic component to their airway disease and stimulated with concanavalin A (Con A) (0.03 to 10.0 micrograms/ml) and anti-IgE (10 to 1,000 ng/ml). Basophil histamine release to Con A and anti-IgE was significantly greater in both allergic and nonallergic asthmatic patients when compared with normal subjects. In contrast, basophil histamine release to the calcium ionophore A23187 was similar in leukocytes from normal subjects and asthmatic patients, suggesting the observed abnormality in secretion may be limited to an IgE-dependent process. To further determine if basophil histamine releasability in asthma correlated to measures of airway reactivity, bronchial provocation with histamine was performed. An inverse correlation was found between the provocative dose of inhaled histamine required to produce a 20% decrease, PD20, in the FEV1 and the leukocyte histamine release to Con A (p less than 0.05) and anti-IgE (p less than 0.05). Thus, we have new evidence that enhanced IgE-dependent release of leukocyte histamine correlates with airway reactivity in asthma. The mechanism of basophil releasability and its relationship to the pathogenesis of airway reactivity in asthma have yet to be established.

Topics: Adult; Antibodies, Anti-Idiotypic; Asthma; Basophils; Bronchi; Bronchial Provocation Tests; Calcimycin; Concanavalin A; Female; Histamine; Humans; Immunization, Passive; Immunoglobulin E; Leukocytes; Male; Reference Values

Pulmonary macrophages from normal subjects and asthmatic patients were examined for the presence of sugar residues on their surface. The technique of bronchoalveolar lavage was employed to obtain cell samples. Ultrastructural and cytophotometric methods were used for studying the patterns of lectin binding by these two groups of macrophages. Three lectins, Concanavalin A (Con A), Wheat germ agglutinin (WGA) and Ricinus communis agglutinin (RCA), were used in this investigation. Pulmonary macrophages from both normal and asthmatic persons revealed a high level of Con A, WGA and RCA binding. The distribution of the electron dense reaction product on the macrophage surfaces was relatively uniform. Quantitative cytophotometric studies showed that the level of binding of Con A by macrophages from both groups was approximately the same. Similar results were obtained with WGA--the difference between macrophages from normal and asthmatic persons was not statistically significant. In the case of RCA, macrophages from asthmatic patients showed a higher level of lectin binding than macrophages from normal persons. The conclusion is made that macrophages from asthmatic persons have more D-galactose residues on their surface.

Topics: Adolescent; Adult; Aged; Asthma; Concanavalin A; Humans; Lectins; Lung; Macrophages; Male; Middle Aged; Photomicrography; Plant Lectins; Wheat Germ Agglutinins

Extrinsic asthmatic patients have been reported to have a deficiency of concanavalin A (Con A)-induced suppressor cell function. We tested whether in vitro colchicine and oral theophylline can correct this immunoregulatory abnormality. Asthmatic patients' mononuclear cells were incubated with Con A and/or colchicine and then suppression of proliferation was measured by coculture of these cells with healthy volunteers' mononuclear cells and phytohaemagglutinin. The Con A induced suppressor cell function of 29 theophylline treated patients (26 +/- 16%, mean +/- s.d.) was significantly (P less than 0.002) increased as compared to 21 untreated patients (12 +/- 10%) but significantly (P less than 0.01) decreased as compared to 45 healthy volunteers (39 +/- 17%). A pharmacological concentration (10(-8) M) of colchicine had no significant effect on Con A-induced suppressor cell function of 19 untreated patients (from 12 +/- 9% to 9 +/- 22%) but significantly (P less than 0.05) increased Con A-induced suppressor cell function of 20 theophylline treated patients (from 26 +/- 17% to 36 +/- 19%). Thus asthmatic patients have decreased Con A-induced suppressor cell function which is partially corrected by oral theophylline and almost completely corrected by oral theophylline plus in vitro colchicine. This synergistic effect raises the possibility that oral colchicine together with theophylline may be useful in treating patients with extrinsic asthma.

Topics: Adolescent; Adult; Asthma; Cells, Cultured; Colchicine; Concanavalin A; Female; Humans; Male; Middle Aged; Mitosis; T-Lymphocytes, Regulatory; Theophylline

Concanavalin-A (Con-A)-induced suppressor activity against the proliferative response of autologous lymphocytes to phytohaemagglutinin (PHA) was examined in the peripheral-blood lymphocytes from fourteen patients with bronchial asthma, ten patients with allergic rhinitis and eleven patients with atopic dermatitis and compared with eleven simultaneously studied healthy normals. Eight of fourteen patients (57%) with bronchial asthma, eight of ten patients (80%) with allergic rhinitis and five of eleven patients (45%) with atopic dermatitis demonstrated deficient Con-A-induced suppressor function. Abnormal suppressor-cell functions could play an important role in the pathogenesis of atopic states.

Topics: Asthma; Concanavalin A; Dermatitis, Atopic; Humans; Immunoglobulin E; In Vitro Techniques; Lymphocyte Activation; Phytohemagglutinins; Rhinitis; T-Lymphocytes, Regulatory

The effect of Fanetizole mesylate or CP-48,810, a new immunostimulating drug, on suppressor cell function and IgE synthesis in vitro was evaluated in atopic patients with allergic rhinitis and/or asthma and eczema. In the absence of the drug, histamine (10(-3)M) stimulated blood mononuclear cells from 23 atopic patients suppressed concanavalin A-induced lymphocyte proliferation by a mean (+/- S.E.M.) of 9.3% +/- 3.5 (compared to 25.1% +/- 2.7 for histamine stimulated mononuclear cells from non-atopic controls). The addition of the drug (2.5 X 10(-4)M) in vitro significantly increased histamine suppressor cell activity of atopic patients to 26.6% +/- 3.9 (compared to 24.7% +/- 2.8 for control cells in the presence of the drug). In order to determine a possible mechanism through which CP-48,810 might enhance histamine-induced suppressor activity, we examined the effects of the drug on the production of histamine-induced suppressor factor (HSF) by lymphocytes and the production of prostaglandin E2 by blood monocytes in the presence of HSF. Supernatants generated from histamine (10(-4)M) stimulated patient lymphocytes caused a 9.0% +/- 1.8 suppression of concanavalin A-induced lymphocyte proliferation (compared to 25.0% +/- 3.1 caused by supernatants from normal subjects). If the drug (2.5 X 10(-4)M) was added at the beginning of culture, HSF activity in supernatants derived from atopic lymphocytes increased significantly to 20.2% +/- 1.8 (compared to 23.3% +/- 3.9 for drug treated control supernatants). Prostaglandin E2 production by atopic monocytes exposed to HSF was less than that of normal monocytes in the absence of the drug.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adjuvants, Immunologic; Adolescent; Adult; Asthma; Cells, Cultured; Child; Concanavalin A; Dermatitis, Atopic; Dinoprostone; Eczema; Female; Histamine; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; Lymphocyte Activation; Male; Middle Aged; Monocytes; Prostaglandins E; Rhinitis; T-Lymphocytes, Regulatory; Thiazoles

Concanavalin A (Con A)-induced suppressor action against the proliferative response of autologous lymphocytes to phytohemagglutinin (PHA) was examined in the peripheral blood lymphocytes of 32 patients with bronchial asthma, 10 patients with allergic rhinitis and then compared with 12 simultaneously studied healthy normals. Twenty-two of 32 patients (68.8%) with bronchial asthma, 8 of 10 patients (80%) with allergic rhinitis demonstrated deficient Con-A-induced suppressor function. Abnormal suppressor cell functions could play an important role in the pathogenesis of respiratory allergic states.

Topics: Adolescent; Adult; Asthma; Child; Child, Preschool; Concanavalin A; Female; Humans; Immunoglobulin E; Male; Middle Aged; Rhinitis, Allergic, Perennial; T-Lymphocytes, Regulatory

In 8 allergic asthmatic patients and 12 healthy volunteers, we investigated the effects of alveolar macrophages (AM) on lymphoproliferative responses to polyclonal T-cell mitogens of autologous peripheral blood mononuclear cells (PBMC). The AM were obtained by bronchoalveolar lavage. Peripheral blood mononuclear cells (PBMC) and mitomycin-treated autologous bronchoalveolar cells (BAC) were cocultured at various AM-to-PBMC ratios, and were stimulated or not by phytohemagglutinin and concanavalin A. Half of AM expressed HLA-DR antigens in both the asthmatic and the control group. The BAC from normal subjects were able to modify the lymphoproliferative responses of autologous PBMC to cell mitogens. A dose-dependent effect was observed related to the number of BAC added to PBMC--suppressive at high ratios but enhancing at low ratios. In allergic bronchial asthma, there was a decreased suppressor cell activity of BAC. Among BAC the adherent cells were responsible for this effect. This abnormality could be a part of more general decreased functional activity of AM in allergic bronchial asthma.

Topics: Adolescent; Adult; Asthma; Cells, Cultured; Concanavalin A; Female; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Lymphocyte Activation; Macrophages; Male; Middle Aged; Monocytes; Phytohemagglutinins; Pulmonary Alveoli

The influence of oxyfedrine (beta-adrenoagonist) and carbocholine (cholinoagonist) on the functional activity of Con A-induced suppressor T-lymphocytes in healthy persons and in bronchial asthma patients was studied. Oxyfedrine at a concentration of 10 micrograms/ml (10(-5) M) was shown to induce a significant increase in the suppressing activity of both normal lymphocytes and those obtained from bronchial asthma patients. The repeated incubation of lymphocytes with carbocholine at a concentration of 5 micrograms/ml (10(-6) M) led to the removal of the suppressing effect of normal lymphocytes and to the increase of the activating effect of lymphocytes from bronchiae asthma patients.

Topics: Asthma; Carbachol; Cells, Cultured; Concanavalin A; Humans; Lymphocyte Activation; Oxyfedrine; Propiophenones; T-Lymphocytes, Regulatory

We have examined in vitro cell-mediated lymphocyte responses to Concanavalin A, (Con. A), and the effects of histamine and indomethacin upon these responses, in normal subjects, and patients with extrinsic and intrinsic asthma, and chronic bronchitis. Lymphocytes from both intrinsic and extrinsic asthmatics are particularly sensitive to histamine-induced suppression of their response to Con. A, and this increased sensitivity was reversed by indomethacin. In these respects, lymphocytes from intrinsic and extrinsic asthmatics behave in an identical fashion, but differ significantly from lymphocytes from both normal subjects and patients with fixed airways obstruction (chronic bronchitis). It is suggested that there is a common immunological mechanism in extrinsic asthma and intrinsic asthma.

Topics: Adult; Aged; Asthma; Bronchitis; Concanavalin A; Humans; Indomethacin; Lymphocyte Activation; Middle Aged

The authors studied the distribution of T-lymphocytes (T-Et and T-Ea) and the nonspecific T-suppressor activity induced by ConA in a group of 104 patients (adults and children) with extrinsic asthma. Comparison with the control group revealed a diminution in the T-Et population (p less than 0.01), no significant change in the T-Ea population and a diminished nonspecific T-suppressor activity (p less than 0.001). The possible role of the T functional deficiency in the development of allergic diathesis is discussed.

Topics: Adolescent; Adult; Asthma; Child; Child, Preschool; Concanavalin A; Female; Humans; Leukocyte Count; Lymphocyte Activation; Lymphokines; Male; Suppressor Factors, Immunologic; T-Lymphocytes

Several immunologic measurements were evaluated in 128 asthmatic patients (81 pediatric and 47 adults) in an effort to examine the question of altered cell-mediated immunity in asthmatics. Both pediatric and adult patients showed alterations in some but not all measurements. The results indicate that there are differences among subgroups of asthmatics and in some cases between pediatric and adult patients.

Topics: Adolescent; Adult; Antigens, Bacterial; Asthma; Beclomethasone; Child; Child, Preschool; Concanavalin A; Humans; Immunity, Cellular; Immunoglobulin A; Immunoglobulin E; Leukocyte Count; Lymphocytes; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; Rosette Formation; Skin Tests; Streptodornase and Streptokinase; Tetanus Toxoid

The role of general and suppressor T-cell function was investigated in a group of twelve children with atopic asthma and ten non-atopic control children. Studies of active E rosettes, lymphocyte stimulation, and delayed type skin responsiveness revealed no statistically significant group differences. Data obtained employing a concanavalin A-induced, suppressor T-cell assay revealed that the asthmatics generated significantly less suppressor cell activity than did the normal control group. There was no correlation between lack of suppressor cell function and serum IgE levels. The results of this study support the concept of suppressor T-cell dysfunction in atopic disease.

Topics: Asthma; Child; Concanavalin A; Female; Humans; Immunity, Cellular; Immunoglobulin E; Male; Phytohemagglutinins; Rosette Formation; Skin Tests; T-Lymphocytes, Regulatory; Theophylline

The effects of cyclophosphamide (200 microgram/ml), levamisole (1 microgram/ml) and Concanavalin A (10 microgram/ml) on the in vitro IgE biosynthesis of lymphocytes from 2 allergic children and six healthy adults were studied. The capability of the IgE biosynthesis was enhanced by cyclophosphamide in nine allergic children, suppressed by levamisole in three and not changed by Concanavalin A. The three patients who showed in vitro suppressed IgE synthesis by levamisole had been treated with levamisole for more than three months but only one revealed decreased serum IgE.

Topics: Adolescent; Asthma; Cells, Cultured; Child; Concanavalin A; Cyclophosphamide; Eczema; Female; Humans; Immunoglobulin E; Levamisole; Lymphocytes; Male; T-Lymphocytes

The T-cell regulations of IgG and IgE biosynthesis were studied by treating purified T-cells from normal adult and asthmatic patients with various concentrations of concanavalin A (Con A) and the co-cultivated with allogeneic mononuclear cells in the presence of pokeweed mitogen for seven days. After being treated with 20 microgram/ml or more of Con A for 18 hours, T-cells from both normal and allergic individuals were able to suppress IgG, but not IgE, production of allogeneic lymphocytes. The results suggest that the T-cell regulation of IgE production is different from that of IgG synthesis and allergic patients do retain suppressor T-cell function(s).

Topics: Adolescent; Asthma; Child; Child, Preschool; Concanavalin A; Female; Humans; Immunoglobulin E; Immunoglobulin G; Lymphocyte Activation; Male; T-Lymphocytes

A technically simple procedure is described for simultaneous determination of numbers and functions of human blood lymphocyte subpopulations. The method is based on stimulation with mitogens (PHA, Con A and PWM) with simultaneous rosetting to detect E receptors and C3 receptors. Con A and PWM activate the affinity of C3 receptors in normal lymphocytes increasing the number of cells rosetting with zymosan-complement particles, and also the number of cells giving rosettes with both zymosan-complement particles and sheep red blood cells (B cells and 'D' cells respectively). PHA activates E receptors, increasing the number of T cells detected. The lymphocytes of 33 patients with allergic diseases of the skin and respiratory tract showed a decrease in C3 rosettes during stimulation with Con A and PWM, which suggests a defect of helper and suppressor functions of lymphocytes in these patients.

Topics: Asthma; B-Lymphocytes; Berylliosis; Concanavalin A; Eczema; Humans; Leukocyte Count; Lymphocyte Activation; Mitogens; Phytohemagglutinins; Pokeweed Mitogens; Rosette Formation; T-Lymphocytes

Various polymorphonuclear leukocyte (PMN) functions are dependent on an intact intracellular cytoskeleton consisting of the microtubules and the microfilaments. To investigate the microtublule system in PMNs we observed the spontaneous, Colchicine and Diamide induced cap-formation by fluorescence microscopy ion PMNs obtained from children with bacterial and viral infections demonstrated with 47 +/- 1% a significantly increased number of spontaneous capped PMNs compared to 22 +/- 1% capped cells obtained from controls. Furthermore, 52 +/- 2% PMNs of patients on immunosuppressive therapy exhibited spontaneous surface capping. There was no significant elevation in the number of capped PMNs (30 +/- 2%) obtained from children with viral infections. Colchicine and Diamide increased the number of capped cells in control PMNs as well as in PMNs from patients to 69 +/- 1% and 67 +/- 1%, respectively. Since the increased spontaneous cap formation in PMNs is associated with a defect of microtubule assembly, the various leukocyte function defects described in patients with bacterial infections, bronchial asthma or on immunosuppressive therapy may have to be considered the consequence of an altered microtubule system.

Topics: Asthma; Bacterial Infections; Child; Colchicine; Concanavalin A; Cytoskeleton; Diamide; Humans; Immunosuppression Therapy; Leukocytes; Microtubules; Receptors, Concanavalin A; Virus Diseases

Concanavalin A (Con A)-induced histamine release from cells of subjects with extrinsic asthma, intrinsic asthma or urticaria and normal individuals was examined utilizing a single isotopic enzymatic assay for histamine. Maximum histamine release by Con A occurred with 0.9 to 4.5 microgram/ml. The mean percentage of maximum histamine release by Con A from cells of donors with extrinsic asthma was 36(+/- 19.3)% while that from cells of normal individuals was 21.4(+/- 23.7)%. However, there was no significant difference between the two groups. The higher reactivity to Con A of cells from individuals with intrinsic asthma or urticaria was not observed compared to that of the normal group. The histamine release by Con A was not correlated with IgE level in the plasma (r = 0.35). It was observed that compound 48/80 inhibited the enzymatic reaction in the isotopic, enzymatic assay.

Topics: Asthma; Concanavalin A; Histamine N-Methyltransferase; Histamine Release; Humans; Immunoglobulin E; Urticaria

Lymphocytes from sixteen patients with extrinsic bronchial asthma and sixteen normals were investigated for their in vitro reactivity to stimulation with phytohaemagglutinin (PHA) and concanavalin A (Con A). No significant differences in maximum lymphocyte responses to PHA or Con A, either in the presence of foetal calf serum or autologous plasma, were found. The significance of these results in relation to T suppressor cell control of IgE formation is discussed.

Topics: Adolescent; Adult; Asthma; Concanavalin A; Female; Humans; Lymphocyte Activation; Male; Phytohemagglutinins