calcimycin and Dermatitis--Atopic

Atopic dermatitis (AD) is multifactorial disease that is highly involved in the activity of T cells from the skin lesion. Seeds of Helianthus annuus extract have been traditionally used as anti-inflammatory reagent but few studies have been reported on leaf of H. annuus that are discarded uselessly as an immunomodulator.. Therefore, here, the regulatory effect of Helianthus annuus extract (HAE) on AD via suppression of T cell activity was investigated.. The efficacy of HAE was evaluated in T cells stimulated with CD3/CD28 antibody and PMA/A23187. And demonstration of the alleviating effect of HAE on AD in the ears of Balb/c female mice stimulated with mite extract and DNCB.. Pre-treatment with HAE abrogates IL-2 production from activated T cells. It was also found that HAE suppresses the expression of surface molecules in activated T cells. Cell viability results demonstrated that HAE is not associated with cytotoxicity in resting and activated T cells. Besides, we exhibited that regulated phosphorylation of MAPK through TAK1-IKKα-NFκB by pre-treatment with HAE leads to the suppressive effect of HAE on T cell activation. Oral administration of HAE attenuates manifestations of AD including reduced thickness of dermis and epidermis, decreased IgE level in serum, and declined mRNA levels of atopic cytokines on ear tissues. The ameliorative effect of HAE on AD was found to be associated with suppressed activity of T cells from draining lymph nodes.. Therefore, our results provide that HAE alleviates AD symptoms via modulation of T cell activity. In addition, these results suggest the immunomodulatory effect of HAE on T-cell mediated diseases.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Calcimycin; CD28 Antigens; Cytokines; Dermatitis, Atopic; Dinitrochlorobenzene; Female; Helianthus; I-kappa B Kinase; Immunoglobulin E; Immunologic Factors; Interleukin-2; Mice; Mice, Inbred BALB C; Plant Extracts; RNA, Messenger; Skin; T-Lymphocytes

Mast cells play a very important role in skin allergy and inflammation, including atopic dermatitis and psoriasis. In the past, it was found that neferine has anti-inflammatory and anti-aging effects on the skin, but its effect on mast cells has not yet been studied in detail. In this study, we used mast cells (RBL-2H3 cells) and mouse models to study the anti-allergic and inflammatory effects of neferine. First, we found that neferine inhibits the degranulation of mast cells and the expression of cytokines. In addition, we observed that when mast cells were stimulated by A23187/phorbol 12-myristate-13-acetate (PMA), the elevation of intracellular calcium was inhibited by neferine. The phosphorylation of the MAPK/NF-κB pathway is also reduced by pretreatment of neferine. The results of in vivo studies show that neferine can improve the appearance of dermatitis and mast cell infiltration caused by dinitrochlorobenzene (DNCB). Moreover, the expressions of barrier proteins in the skin are also restored. Finally, it was found that neferine can reduce the scratching behavior caused by compound 48/80. Taken together, our results indicate that neferine is a very good anti-allergic and anti-inflammatory natural product. Its effect on mast cells contributes to its pharmacological mechanism.

Topics: Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Benzylisoquinolines; Calcimycin; Calcium; Cell Line; Cell Movement; Cytokines; Dermatitis, Atopic; Dinitrochlorobenzene; Disease Models, Animal; Mast Cells; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; NF-kappa B; Phosphorylation; Signal Transduction

AST2017-01 mainly consists of Rumex crispus and -Cordyceps militaris and has been widely consumed as an herbal medicine or functional food in Korea. Here we investigated the influences of AST2017-01 and its active component, chrysophanol on human mast cell (HMC-1 cell) and human keratinocyte (HaCaT cell)-mediated inflammatory reactions. Pretreatment with AST2017-01 or chrysophanol suppressed intracellular calcium levels and histamine release in phorbol 12-myristate 13-acetate and calcium ionophore A23187 (PMACI)-treated HMC-1 cells. Levels of phosphorylated-mitogen-activated protein kinase increased by PMACI stimulation were reduced by AST2017-01 or chrysophanol pretreatment. Protein levels of IκB kinaseβ and receptor-interacting protein 2 in PMACI-treated HMC-1 cells were decreased by AST2017-01 or chrysophanol pretreatment. Pretreatment with AST2017-01 or chrysophanol significantly blocked PMACI-induced activation of caspase-1 and nuclear factor-κB. In addition, pretreatment with AST2017-01 or chrysophanol significantly decreased the PMACI-induced levels of interleukin (IL)-1β, IL-6, tumor necrosis factor-α, and thymic stromal lymphopoietin (TSLP) on HMC-1 cells. In activated HaCaT cells, pretreatment with AST2017-01 or chrysophanol significantly reduced production of TSLP and activation of caspase-1. In conclusion, these findings indicate that chrysophanol is an active component of AST2017-01 and AST2017-01 acts as a novel potent anti-inflammatory herbal medicine or functional food.

Topics: Anthraquinones; Anti-Inflammatory Agents; Calcimycin; Calcium Signaling; Caspase 1; Cell Line; Cytokines; Deoxyribonucleases, Type II Site-Specific; Dermatitis, Atopic; Herbal Medicine; Humans; Keratinocytes; NF-kappa B; Rumex; Signal Transduction

Cynanchum atratum Bunge (Apocynaceae) is a folk medicine to treat skin inflammatory diseases. However, the effects of C. atratum on atopic dermatitis have not been elucidated. In this study, we evaluate the effects of aqueous extract of C. atratum (CA) and its molecular mechanism on atopic dermatitis (AD). 1 and 100mg/mL CA were topically applied to 2,4-dinitrochlorobenzene (DNCB)-induced AD-like skin lesions for 11 days. The number of scratching behavior was evaluated for 20min. AD-like symptoms including elevated serum IgE, skin hyperplasia and mast cell infiltration were investigated. The expressions of pro-inflammatory cytokines and mediators were analyzed in AD-like skin legions. In addition, pro-inflammatory cytokine production was confirmed in human mast cells (HMC)-1 stimulated with PMA plus A23187 (PMACI). Topical application of CA attenuated total serum IgE level and scratching behavior. Skin hyperplasia including epidermis and dermis was ameliorated in CA-treated skin legions. The number of infiltrated mast cells was significantly decreased by CA treatment. In addition, CA reduced pro-inflammatory cytokines, such as IL-6, IL-1β and TNF-α and Th2 cytokine, IL-4, in both of AD-like skin lesions and PMACI-sensitized HMC-1 cells. Furthermore, CA decreased the expressions of NF-κB, phospho-IκBα and MAP kinase. These results suggest the inhibitory effects of CA on the development of AD by regulating pro-inflammatory cytokines and mediators. CA could be an effective substance for the treatment of AD.

Topics: Animals; Calcimycin; Cytokines; Dermatitis, Atopic; Dermis; Dinitrochlorobenzene; Epidermis; Female; Hyperplasia; Immunoglobulin E; Inflammation; Interleukin-6; Mast Cells; Mice; Mice, Inbred BALB C; NF-kappa B; NF-KappaB Inhibitor alpha; Plant Extracts; Tumor Necrosis Factor-alpha; Vincetoxicum

Atopic dermatitis (AD) is one of the most common inflammatory cutaneous diseases. Thymic stromal lymphopoietin (TSLP) has been demonstrated to be an important immunologic factor in the pathogenesis of AD. The production of TSLP can be induced by a high level of intracellular calcium concentration and activation of the receptor-interacting protein 2/caspase-1/NF-κB pathway. Andrographolide (ANDRO), a natural bicyclic diterpenoid lactone, has been found to exert anti-inflammatory effects in gastrointestinal inflammatory disorders through suppressing the NF-κB pathway.. To explore the effect of ANDRO on the production of TSLP in human mast cells and AD mice model.. We utilized enzyme-linked immunosorbent assay, real-time reverse transcription polymerase chain reaction analysis, Western blot analysis, and immunofluorescence staining assay to investigate the effects of ANDRO on AD.. ANDRO ameliorated the increase in the intracellular calcium, protein, and messenger RNA levels of TSLP induced by phorbol myristate acetate/calcium ionophore A23187, through the blocking of the receptor-interacting protein 2/caspase-1/NF-κB pathway in human mast cell line 1 cells. ANDRO, via oral or local administration, also attenuated clinical symptoms in 2,4-dinitrofluorobenzene-induced AD mice model and suppressed the levels of TSLP in lesional skin.. Taken together, ANDRO may be a potential therapeutic agent for AD through suppressing the expression of TSLP.

Topics: Animals; Calcimycin; Cell Proliferation; Cell Survival; Cytokines; Dermatitis, Atopic; Dinitrofluorobenzene; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Humans; Mast Cells; Mice; Molecular Conformation; Structure-Activity Relationship; Tetradecanoylphorbol Acetate; Thymic Stromal Lymphopoietin; Tumor Cells, Cultured

Naju Jjok (NJJ, Polygonum tinctorium) is a clear heat and release toxin medicinal. It has been used to treat various inflammatory diseases and as a dye in clothing in traditional Korean medicine. However, the effect of NJJ on atopic dermatitis (AD) has not been elucidated. Therefore, we examined whether NJJ would have an inhibitory effect on AD using the mimic AD murine model and in vitro model.. We treated NJJ on 2,4-dinitrofluorobenzene (DNFB)-induced AD-like skin lesions in NC/Nga mice, phorbol myristate acetate/calcium ionophore A23187-stimulated human mast cell line (HMC-1) cells, and anti-CD3/anti-CD28-stimulated splenocytes. Histological analysis, ELISA, PCR, and Western blot analysis were performed.. The oral administration with NJJ suppressed the total clinical severity in DNFB-induced AD-like lesional skin. NJJ significantly suppressed the levels of inflammatory mRNA and protein in AD-like lesional skin. NJJ significantly suppressed the levels of IgE and interleukin-4 in the serum of DNFB-induced AD mice. The expression of mast cells-derived caspase-1 was suppressed by NJJ in AD-like lesional skin. In addition, topical application with NJJ improved clinical symptoms in DNFB-induced AD mice. The topical application with NJJ significantly suppressed the levels of IgE and histamine in the serum of DNFB-induced AD mice. NJJ suppressed the production and mRNA expression of TSLP by blockade of caspase-1 signal pathway in the activated HMC-1 cells. Furthermore, NJJ significantly decreased the production of tumor necrosis factor-α from the stimulated splenocytes.. In conclusion, these results propose curative potential of natural dye, NJJ by showing the scientific evidence on anti-AD effect of NJJ which has been used traditionally.

Topics: Administration, Oral; Animals; Calcimycin; Caspase 1; Cell Line; Cytokines; Dermatitis, Atopic; Dinitrofluorobenzene; Drug Evaluation, Preclinical; Humans; Male; Medicine, Korean Traditional; Mice; Phytotherapy; Plant Extracts; Polygonum; Receptor-Interacting Protein Serine-Threonine Kinase 2; Tetradecanoylphorbol Acetate; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor-alpha

Atopic dermatitis (AD) is a common skin disease that greatly worsens quality of life. Thymic stromal lymphopoietin (TSLP) plays a decisive role in the development of AD. The purpose of this study is to examine whether tryptanthrin (TR) would suppress AD through the regulation of TSLP. We analyzed the effect of TR on the level of TSLP from phorbol myristate acetate/calcium ionophore A23187-activated human mast cell line, HMC-1 cells, in 2,4-dinitrofluorobenzene-induced AD-like skin lesions of NC/Nga mice, and in anti-CD3/anti-CD28-stimulated splenocytes. TR significantly suppressed the level of intracellular calcium and the production and mRNA expression of TSLP through the blockade of receptor-interacting protein 2/caspase-1/nuclear factor-κB pathway in the activated HMC-1 cells. TR also significantly suppressed the levels of histidine decarboxylase and IL-1β. Furthermore, TR ameliorated clinical symptoms in the AD model. TR significantly reduced the levels of TSLP, IL-4, IFN-γ, IL-6, TNF-α, thymus and activation-regulated chemokine, and caspase-1 in AD skin lesions. Also, TR significantly reduced the serum levels of histamine and IL-4 in the AD model. Finally, TR significantly inhibited the production of IL-4, IFN-γ, and TNF-α from the stimulated splenocytes. Taken together, TR exhibits the potential to be a therapeutic agent for AD through down-regulation of TSLP.

Topics: Animals; Calcimycin; Calcium Ionophores; Cell Line; Cytokines; Dermatitis, Atopic; Dinitrofluorobenzene; Down-Regulation; Humans; Male; Mast Cells; Mice; Quinazolines; Skin; Spleen; Tetradecanoylphorbol Acetate; Thymic Stromal Lymphopoietin

Sophoricoside exhibits numerous pharmacological effects, including anti- inflammatory and anti-cancer actions, yet the exact mechanism that accounts for the anti-allergic effects of sophoricoside is not completely understood. The aim of the present study was to elucidate whether and how sophoricoside modulates the mast cell-mediated allergic inflammation in vitro and in vivo. We investigated the pharmacological effects of sophoricoside on both compound 48/80 or histamine-induced scratching behaviors and 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis in mice. Additionally, to find a possible explanation for the anti-inflammatory effects of sophoricoside, we evaluated the effects of sophoricoside on the production of histamine and inflammatory cytokines and activation of nuclear factor-κB (NF-κB) and caspase-1 in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated human mast cells (HMC-1). The finding of this study demonstrated that sophoricoside reduced compound 48/80 or histamine-induced scratching behaviors and DNCB-induced atopic dermatitis in mice. Additionally, sophoricoside inhibited the production of inflammatory cytokines as well as the activation of NF-κB and caspase-1 in stimulated HMC-1. Collectively, the findings of this study provide us with novel insights into the pharmacological actions of sophoricoside as a potential molecule for use in the treatment of allergic inflammation diseases.

Topics: Animals; Anti-Inflammatory Agents; Benzopyrans; Calcimycin; Calcium Ionophores; Carcinogens; Caspase 1; Cell Line; Cytokines; Dermatitis, Atopic; Dinitrochlorobenzene; Histamine; Humans; Irritants; Male; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; NF-kappa B; Tetradecanoylphorbol Acetate

Although clinical evidence has suggested that dysregulated fatty acid metabolism is associated with atopic disorders, the molecular basis for such a correlation remains to be demonstrated. In the present study, we analyzed the fatty acid composition in peripheral blood cells of NC/Nga mice, a model for atopic dermatitis (AD). We found that arachidonic acid significantly accumulated in mice with the AD manifestation. In addition, the leucotriene B4-releasing ability upon calcium ionophore A23187 stimulation was potentiated in blood cells. An arachidonic acid accumulation was not apparent in the non-atopic BALB/c strain, but was still observed in healthy NC/Nga mice fed under specific pathogen-free conditions. These results indicate that a disturbed fatty acid metabolism in NC/Nga mice was not a trigger factor for their dermatitis development.

Topics: alpha-Linolenic Acid; Animals; Arachidonic Acid; Blood Cells; Calcimycin; Dermatitis, Atopic; Disease Models, Animal; Fatty Acids; Humans; In Vitro Techniques; Ionophores; Leukotriene B4; Male; Mice; Mice, Inbred BALB C; Mice, Mutant Strains

Because of the implication of histamine in canine atopic dermatitis, H1-antihistamines may provide a valid alternative to glucocorticoid therapy. In vitro study of these drugs prior to clinical testing can allow the most promising compounds to be selected for trials and render trials with drugs of doubtful efficacy unnecessary.. Isolated canine cutaneous mast cells.. Cells were preincubated with antihistamines at increasing concentrations and incubated with concanavalin A (1,000 micrograms/ml), calcium ionophore A23187 (1 microM), and substance P (100 microM). Compound 48/80 was not used because it proved to be cytotoxic.. Generally, significant prodegranulating effect was not observed for most of the studied agents. Only terfenadine increased spontaneous histamine release at concentrations > 30 microM. Cetirizine did not block histamine release at any of the studied concentrations. Ketotifen had a low inhibitory effect only at the highest concentration (100 microM) after concanavalin A- (23.6 +/- 2.8%) and calcium ionophore A23187- (29.8 +/- 3.0%) induced release. Terfenadine caused a concentration-dependent inhibitory effect after ionophore A23187- (48.1 +/- 2.2%) and concanavalin A- (28.9 +/- 2.3%) activation, but was inactive against substance P-induced release. In contrast, loratadine had potent dose-dependent inhibition of concanavalin A- and ionophore A23187-induced histamine release, with maximal effect of 85.6 +/- 3.1% and 62.6 +/- 4.7%, respectively, at 100 microM concentration. After substance P activation, histamine release was only slightly inhibited by loratadine (14.8 +/- 1.1%).. This study documents the behavior of isolated canine cutaneous mast cells in the presence of nonimmunologic stimulation. Using this in vitro method, we were able to determine that loratadine is the only antihistamine that has potent inhibition of histamine release from dog cutaneous mast cells without a substantial prodegranulating effect. Loratadine is, therefore, a good candidate for clinical testing.

Topics: Animals; Calcimycin; Cell Survival; Cetirizine; Concanavalin A; Dermatitis, Atopic; Dog Diseases; Dogs; Dose-Response Relationship, Drug; Histamine H1 Antagonists; Histamine Release; In Vitro Techniques; Ketotifen; Loratadine; Mast Cells; Skin; Substance P; Terfenadine

Isolated dermal mast cells from atopic dogs are a valuable tool for the analysis of their functional properties in atopic dermatitis. We have characterized the histamine secretory pattern of mast cells enzymatically dispersed from the skin of dogs naturally suffering from this condition. The total histamine content found per isolated skin mast cell was higher in the allergic dogs than in nonatopic (control) animals (8.7 pg/mast cell versus 5.2 pg/mast cell). This phenomenon together with the well known higher concentration of skin mast cell number in atopic dermatitis lesions might account for the observed increase in local histamine concentration (15.0 micrograms/g versus 9.0 micrograms/g). Atopic dog-derived mast cells were highly reactive to both non-immunological (ionophore A23187) and an immunological-like (concanavalin A) stimulus. Furthermore, histamine net release induced by concanavalin A (1 mg/ml) stimulation was clearly enhanced in the atopic dogs (33.3% net release versus 15.4% in controls). These results have not been described in dermal mast cells dispersed from the skin of individuals with atopic dermatitis and clearly support the hypothesis that mast cells play a major role in causing and possibly modulating atopic dermatitis, through enhanced sensitivity or releasability. However, whether these two phenomena are primary abnormalities of atopic dermatitis, or only secondary changes, remains undetermined.

Topics: Animals; Calcimycin; Cell Count; Concanavalin A; Dermatitis, Atopic; Dogs; Histamine; Histamine Release; Ionophores; Mast Cells; Skin

The ex vivo release of leukotrienes B4 (LTB4) and C4 (LTC4) from the leukocytes of children with atopic dermatitis (AD) was evaluated after stimulation with Ca-ionophore and opsonized zymosan and compared with that of control children of similar ages. The blood eosinophil counts and total serum IgE levels in AD children were significantly higher than those in control children. The production of LTC4, but not LTB4, was significantly higher in AD children than in control children. There was a significant correlation between the relative blood eosinophil count and LTC4 generation after stimulation with both Ca-ionophore and opsonized zymosan in all subjects. Calculations of the amount of LTC4 produced per eosinophilic cell showed that there was no significant difference between cells from AD children and control children in terms of their ability to produce LTC4. These findings suggest that the enhanced LTC4 generation is due to increased numbers of eosinophils rather than to enhanced releasability of these cells.

Topics: Adolescent; Calcimycin; Child; Child, Preschool; Dermatitis, Atopic; Eosinophils; Female; Humans; Leukocyte Count; Leukocytes; Leukotriene B4; Leukotriene C4; Male; Zymosan

In view of the high incidence of canine cutaneous atopic disease and the relevance of mast cells to its pathogenesis, it was considered important to isolate firstly cutaneous mast cells from normal dog skin and to assess the histamine secretory activity, as this can be further used as a tool for the study of canine skin mast cell pharmacology in cutaneous atopy. The procedure for canine dermal mast cell dispersion following a skin enzymatic digestion (as for previous human skin mast cell dispersion methods) is described in detail. The number of canine cutaneous mast cells yielded per gram of skin was 2.31 +/- 0.21 x 10(5) representing 1.00% of the total cutaneous cells. The total histamine content per mast cell is 4.93 +/- 0.39 pg. Net histamine release owing to stimulation by calcium ionophore A23187 (1 microM) and concanavalin A (1 mg ml-1) was respectively 32.17 +/- 3.56% and 20.39 +/- 2.41% of the total amount per cell. Viability and reactivity to both stimuli of dispersed cutaneous mast cells were similar to the results found in humans. The present study allows further research on the role of mast cells immunopharmacology in allergy by investigation of cells isolated from canine skin in naturally occurring or experimentally induced atopy in the dog to be undertaken.

Topics: Animals; Calcimycin; Cell Count; Cell Separation; Cell Survival; Cells, Cultured; Concanavalin A; Dermatitis, Atopic; Dog Diseases; Dogs; Female; Histamine Release; Male; Mast Cells; Skin

The effect of disodium cromoglycate on in vitro proliferative responses of peripheral blood mononuclear cells from healthy individuals, allergic patients with moderate serum IgE and patients with atopic dermatitis and high levels of serum IgE was investigated. Peripheral blood mononuclear cells were stimulated with mitogens (phytohaemagglutinin, Concanavalin A), recombinant interleukin-2, calcium ionophore + phorbol 12-myristate 13-acetate, purified protein derivative of tuberculin and allergens. It was possible to induce in vitro specific, allergen-triggered responses only in allergic individuals with moderate serum IgE and not in individuals with atopic dermatitis and high serum IgE. Generally, whenever the stimulatory signal(s) caused a significant proliferative response, disodium cromoglycate inhibited the proliferation. This inhibition was seen for all activation agents and for both healthy and allergic individuals. By contrast, for certain non- or low-responders (both healthy and allergic individuals) disodium cromoglycate seemed to amplify the proliferation to various activation signals. Only non- or low-responder cells derived from atopic dermatitis patients showed a biphasic kinetic response pattern when stimulated with the drug in combination with recombinant interleukin-2, recombinant interleukin-2 + ionophore or specific allergens.

Topics: Calcimycin; Cells, Cultured; Cromolyn Sodium; Dermatitis, Atopic; Female; Humans; Hypersensitivity; Interleukin-2; Lymphocyte Activation; Male; Recombinant Proteins; Tuberculin

The generation of arachidonic acid-derived inflammatory mediators from unstimulated and stimulated neutrophils (PMN) and platelets in the presence of exogenous LTA4 has been studied in patients with atopic dermatitis (AD) as well as in healthy volunteers. PMN were stimulated with the interleukins IL-3, IL-8, C5a, and the Ca-ionophore A23187. In addition, NaF and thrombin were used to stimulate platelets. The release of leukotriene (LT)B4, 20-COOH- and 20-OH-LTB4, cysteinyl-leukotrienes and 12-HETE was measured. The proinflammatory mediator release from PMN and platelets of patients with AD was significantly higher as compared to the control group. The spontaneous conversion of LTA4 by PMN and platelets was markedly enhanced in patients with AD. Different results with receptor-specific and non-specific stimuli (Ca-ionophore A23187) in the presence of exogenous LTA4 were obtained. The results indicate a higher state of activation for enzymes involved in leukotriene formation. Furthermore, the production of 12-HETE by platelets from patients with AD was enhanced in unstimulated and stimulated cells. Our data emphasize that neutrophils and platelets may play an important role in the pathogenesis of AD by an increased responsiveness to receptor-specific stimuli and cell-cell interaction via LTA4.

Topics: Adolescent; Adult; Blood Platelets; Calcimycin; Cells, Cultured; Complement C5a; Dermatitis, Atopic; Humans; Interleukins; Leukotriene A4; Leukotrienes; Neutrophils

Purified peripheral blood monocytes isolated from patients with atopic dermatitis (AD) and from nonallergic normal donors were compared for their abilities to release leukotriene C4 (LTC4), leukotriene B4 (LTB4) and beta-glucuronidase in response to challenge with aggregated immunoglobulins or anti-immunoglobulins. The relationship between mediator release and the number of monocytes that formed rosettes with immunoglobulin-coated indicator cells was examined. Patients with AD had twice as many IgA- and three times as many IgE-rosetting monocytes as normal donors (48 +/- 12% versus 27 +/- 10% and 40 +/- 15% versus 14 +/- 3%, respectively), and yet the amounts of IgA- and IgE-induced LTC4 released were similar for both groups. This apparent discrepancy did not result from a decreased capacity for arachidonate metabolism via the C5-lipoxygenase pathway, since stimulation of monocytes from patients and normal donors with the calcium ionophore A23187 induced similar amounts of LTC4 and LTB4 release (LTC4, 3.0 +/- 1.7 versus 3.0 +/- 1.0 ng/10(6) cells; LTB4, 5.3 +/- 0.7 versus 5.2 +/- 0.5 ng/10(6) cells, respectively). In addition, aggregated IgG-induced LTC4 release by monocytes of both groups was similar, concomitant with an equivalent number of IgG-rosetting cells. Determination of cytophilically bound IgG and IgE by flow cytometry demonstrated that monocytes from atopic patients had more IgG bound than monocytes from normal donors. Similar amounts of IgE were detected on most monocytes from both groups, despite the higher serum IgE levels of patients. However, approximately 3% to 8% of monocytes from atopic but not normal donors stained brightly for IgE, suggesting that relatively large amounts of cytophilic IgE were bound to a small percentage of the patients' monocytes. Challenge of monocytes with anti-IgE or anti-IgG induced release of similar amounts of LTC4 for both groups, despite the presence of more cytophilic IgG on monocytes from atopic donors. These data indicate that monocytes from patients with AD release LTC4 and LTB4 in response to challenge with aggregated IgE or anti-IgE, as well as aggregated IgG, IgA, and anti-IgG. However, under our in vitro conditions, stimulation of patients' monocytes with aggregated IgA or IgE was not associated with increased mediator release, despite higher percentages of IgA- and IgE-rosetting cells compared to normal donors.

Topics: Antigens, Differentiation; Antigens, Differentiation, B-Lymphocyte; Calcimycin; Dermatitis, Atopic; Female; Fluorescent Antibody Technique; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin Fab Fragments; Immunoglobulin G; Leukotriene B4; Macromolecular Substances; Male; Monocytes; Receptors, Fc; Receptors, IgE; Receptors, IgG; Rosette Formation; SRS-A

The releasability of arachidonic acid-derived inflammatory mediators (eicosanoids) from peripheral blood leukocytes has been tested in patients with atopic eczema and healthy, non-atopic controls. Spontaneous and stimulated release of prostaglandin E2 (PGE2), leukotriene B4 (LTB4) and leukotriene C4 (LTC4) has been measured after challenge of cells with various concentrations of anti-IgE, Ca-ionophore A23187 and C5a. The maximal stimulation of cells with Ca-ionophore resulted in the generation of high amounts of all eicosanoids, which was essentially equal in both atopic and control groups. On the other hand, enhanced spontaneous and stimulated eicosanoid release was noted after immunological challenge using C5a and anti-IgE in the atopic eczema group. Thus, our data support the hypothesis of enhanced releasability of inflammatory mediators in atopic eczema.

Topics: Adolescent; Adult; Aged; Antibodies, Anti-Idiotypic; Arachidonic Acids; Calcimycin; Child; Complement C5; Complement C5a; Dermatitis, Atopic; Dinoprostone; Female; Humans; Immunoglobulin E; Leukocytes; Leukotriene B4; Male; Middle Aged; Prostaglandins E; SRS-A

"Releasability" is the theory whereby biochemical events in basophils influence the capacity to release chemical mediators in response to activating stimuli. We have compared the releasability of basophils from 21 young patients with atopic dermatitis (AD) with that from 17 normal donors of matched ages. Basophils were challenged with several different stimuli: rabbit antihuman Fc epsilon (anti-IgE), N-formyl-methionyl-leucyl-phenylalanine (f-met-peptide), Ca++ ionophore A23187, and D2O. Basophils from patients with AD released significantly more histamine both "spontaneously" and in response to D2O than did controls. The basophils of patients with AD were significantly more responsive to anti-IgE and to A23187. There was no difference between the percent f-met-peptide-induced histamine release in patients with AD vs controls. No significant correlation between percent histamine release with optimal or suboptimal concentrations of the stimuli and serum IgE level was found. There was a significant correlation between the sensitivity of the cells to release with f-met-peptide and the response to A23187 both in control and in AD patients. Since basophils are thought to play some role at the site of inflammation in AD, their increased releasability might contribute to the symptoms of these patients.

Topics: Adolescent; Antibodies, Anti-Idiotypic; Basophils; Calcimycin; Child; Child, Preschool; Dermatitis, Atopic; Deuterium; Deuterium Oxide; Histamine Release; Humans; Immunoglobulin E; Infant; N-Formylmethionine Leucyl-Phenylalanine; Water

Topics: Age Factors; Animals; Antibodies, Anti-Idiotypic; Asthma; Basophils; Calcimycin; Dermatitis, Atopic; Deuterium; Deuterium Oxide; Histamine Release; Humans; Hypersensitivity; Immunoglobulin E; N-Formylmethionine Leucyl-Phenylalanine; Phenotype; Water