To investigate the imbalance of the cytokine production profile of T cells from atopic asthmatics, we measured concentrations of IL-4, IL-5 and IFN-y by ELISA method in the culture supernatants of peripheral blood mononuclear cells(PBMCs) and Derrnato-phagoides pteronyssinus(Der p) J-stimulated PBMCs from Der p-sensitized atopic asthmatics, Der p-sensitized healthy atopits, non-atopic asthmatics and healthy non-atopics. The suppressive effect of IFN-y on cytokine production of Der p J-stimulated PBMCs was also examined. The PBMCs from atopics showed higher IL-4 and IL-5 production in response to PHA +TPA and higher IFN-gamma production in response to Der p Jq compared with non-atopits. The Der p J-stimulated PBMCs from atopics showed a tendency of increased IL-5 production in response to Der p J and higher IL-4 and IL-5 production in response to PHA+TPA compared with non-atopics. IL-5 production of Der p J-stimulated PBMCs from atopics was suppressed by IFN It is suggested that an imbalance in IL-4, IL-5 and IFN-y production is a feature of the atopic state. The TH2 characteristics of allergen-stimulated PBMCs could be regulated by IFN-y.
Enzyme-Linked Immunosorbent Assay ; Interleukin-4 ; Interleukin-5 ; T-Lymphocytes

BACKGROUND: A small subgroup of atopic dermatitis (AD) patients show low total and allergen-specific immunoglobulin (IgE) levels. This subgroup has been termed 'intrinsic' AD (IAD) as compared to its counterpart 'extrinsic' AD (EAD). However, the difference of cytokine expression between IAD and EAD has not been fully understood. OBJECTIVE: To compare the expression of various inflammatory cytokines in the peripheral blood mononuclear cells (PBMCs) and lesional skin of patients with IAD and EAD, which are known to be associated with AD pathophysiology. METHODS: We assessed the protein levels of cytokines in the PBMCs and lesional skin. We evaluated the levels of IL-3, IL-4, IL-5, IL-6, IL-10, IL-13, FcepsilonRI and FcepsilonRII from the PBMCs and lesional skin of patients with IAD and EAD. RESULTS: The patients with EAD had elevated levels of the IL-3 expression in their PBMCs and elevated levels of FcepsilonRI in their lesional skin compared to that of the patients with IAD. The expression of other cytokines did not differ in the PBMCs and lesional skin from the two subgroups. CONCLUSION: This study suggests that IL-3 could be associated with the pathophysiology of EAD as compared to that of IAD, along with FcepsilonRI which was previously shown to be highly expressed in EAD patients.
Cytokines ; Dermatitis, Atopic ; Humans ; Immunoglobulins ; Interleukin-10 ; Interleukin-13 ; Interleukin-3 ; Interleukin-4 ; Interleukin-5 ; Interleukin-6 ; Skin

BACKGROUND: It is well known that the expression of Th2 cytokines are up-regulated in the atopic asthma. The study was to investigate the expression of transcription factors, such as GATA-3, c-maf, T-bet which are known to be involved in the T cell differentiation in the peripheral blood mononuclear cells (PBMCs) of atopic asthmatics. METHODS: PBMCs were obtained from non-atopic controls and atopic asthmatics and cultured for 48 hours, and then 12-o-tetracanoylphorbol-13-acetate (PMA) and calcium ionophore (ionomycin) were added. mRNA of GATA-3, c-maf, T-bet, IL-4, IL-5, IL-13 and IFN-gamma were measured by RT-PCR RESULTS: mRNAs of Th2 cytokines, such as IL-4, IL-5 and IL-13 were expressed higher in the atopic asthmatics, and that of Th1 cytokine, IFN-gamma was expressed higher in the non-atopic controls. GATA-3 and c-maf were expressed higher and T-bet was expressed lower in the atopic asthmatics. Expressions of GATA-3, c-maf and T-bet were not changed with the stimulation of PMA/Ionomycin. CONCLUSION: This study suggest that GATA-3, c-maf and T-bet participate in the Th2 type inflammation in atopic asthmatics, like GATA-3 and c-maf as stimulatory factors, and T-bet as inhibitory factor.
Asthma ; Calcium ; Cell Differentiation ; Cytokines ; Inflammation ; Interleukin-13 ; Interleukin-4 ; Interleukin-5 ; RNA, Messenger* ; Transcription Factors

PURPOSE: Group 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens. METHODS: Flow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage- and Lineage+ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage- and Lineage+ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit. RESULTS: The percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33+/-IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage- cells compared to Lineage+ cells. CONCLUSIONS: AR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.
Allergens ; Artemisia ; Cytokines ; Humans ; Interleukin-13 ; Interleukin-2 ; Interleukin-5 ; Lymphocytes ; Pyroglyphidae ; Rhinitis*

BACKGROUND: It has been well known that bronchial asthma is a chronic inflammatory disorder. The "activation" of lymphocytes has a significant role in the pathogenesis of bronchial asthma. Among these lymphocytes, TH2-like rather than TH1-like lymphoytes are activated in the bronchial tissues from patients with atopic bronchial asthma. However, the difference of cytokines expression is not well documented between the atopic normal subjects and atopic asthmatics. METHODS: Bronchial tissues were obtained from the tweleve atopic and non-atpoic asthmatics and tweleve atopic and non-atopic healthy subjects for in stiu hybridizatin of IL-2, IL-4, IL-5, and INF-gamma. The probe of cytokines were tagged with digoxigenin by random priming method. RESULTS: The infiltration of many inflammatory cells on submucosa and denuded epithelium were observed in the bronchial tissue from patients with bronchial asthma. The RNase-treated bronchial tissues did not have the brown signal on the tissue, but, RNase-untreated bronchial tissues had the positive brown signal on the inflammatory cells under the basement membrane. The IL-2 positive signals were detected in 2 cases, IFN-gamma in 1 case, IL-4 in 2 cases, IL-5 in 2 cases among 6 non-atopic healthy subjects. The atopic healthy subjects showed 1 case of positive signal of IL-2 and IFN-gamma, but did not show any signals of IL-4 and IL-5. The positive signals of IL-2 were detected in 4 cases among 6 atopic and 6 non-atopic asthmatics, 2 cases and 1 case of IFN-gamma respectively, 4 cases and 3 cases of IL-4 respectively, 4 cases and 3 cases of IL-5 respectively. CONCLUSION: The lymphocytes were activated in the bronchus of asthmatics. Among lymphocytes, TH2-like lymphocytes may be involved in the pathogenesis of bronchial asthma. However, futher study with immunohistochemical stain may be necessary for defining the source of cytokines, because of TH2-like lymphocytes were also activated in some atopic healthy subjects.
Asthma* ; Basement Membrane ; Bronchi ; Cytokines* ; Digoxigenin ; Epithelium ; Humans ; Interleukin-2 ; Interleukin-4 ; Interleukin-5 ; Lymphocytes

No abstract available.
Asthma ; Chemotaxis ; Cytokines* ; Eosinophils* ; Interferon-gamma ; Interleukin-5

Several pathophysiologic features of allergic inflammation, such as T(H) differentiation, the regulation of IgE, eosinophilia, mast cell proliferation, and cellular recruitment, are regulated by various cytokines. It has been of particular interest to investigate the underlying mechanism in the preferential activation of T(H2) cells by allergens. Although interleukin (IL)-4 is the major determinant of T(H2) differentiation, the original cellular source of IL-4 and the nature of the interaction between IL-4 and TH2 differentiation remain unclear. Recent studies have demonstrated that the regulation of IgE depends primarily on the functional activities of IL-4, IL-13, and IFN-gamma. Eosinophilia and an increased number of mast cells characterize allergic inflammation, which is a T cell-dependent process. IL-5 is the major chemotactic and activating factor of eosinophils. Mast cell proliferation results from several cytokines, including IL-3, IL-9, and IL-10. It has been suggested that, in the late phase reaction of allergic inflammation, proinflammatory cytokines released from mast cells, eosinophils, and T(H2) cells enhance the expression of adhesion molecules and chemokines that further promote the allergic cellular milieu.
Allergens ; Chemokines ; Cytokines* ; Eosinophilia ; Eosinophils ; Hypersensitivity* ; Immunoglobulin E ; Inflammation ; Interleukin-10 ; Interleukin-13 ; Interleukin-3 ; Interleukin-4 ; Interleukin-5 ; Interleukin-9 ; Interleukins ; Mast Cells

Several pathophysiologic features of allergic inflammation, such as T(H) differentiation, the regulation of IgE, eosinophilia, mast cell proliferation, and cellular recruitment, are regulated by various cytokines. It has been of particular interest to investigate the underlying mechanism in the preferential activation of T(H2) cells by allergens. Although interleukin (IL)-4 is the major determinant of T(H2) differentiation, the original cellular source of IL-4 and the nature of the interaction between IL-4 and TH2 differentiation remain unclear. Recent studies have demonstrated that the regulation of IgE depends primarily on the functional activities of IL-4, IL-13, and IFN-gamma. Eosinophilia and an increased number of mast cells characterize allergic inflammation, which is a T cell-dependent process. IL-5 is the major chemotactic and activating factor of eosinophils. Mast cell proliferation results from several cytokines, including IL-3, IL-9, and IL-10. It has been suggested that, in the late phase reaction of allergic inflammation, proinflammatory cytokines released from mast cells, eosinophils, and T(H2) cells enhance the expression of adhesion molecules and chemokines that further promote the allergic cellular milieu.
Allergens ; Chemokines ; Cytokines* ; Eosinophilia ; Eosinophils ; Hypersensitivity* ; Immunoglobulin E ; Inflammation ; Interleukin-10 ; Interleukin-13 ; Interleukin-3 ; Interleukin-4 ; Interleukin-5 ; Interleukin-9 ; Interleukins ; Mast Cells

Eosinophils have been reported to modulate T cell responses. Previously, we reported that high-mobility group box 1 protein (HMGB1) played a key role in the pathogenesis of asthma. This study was conducted to test our hypothesis that eosinophils could modulate T cell responses via HMGB1 in the pathogenesis of asthma characterized by eosinophilic airway inflammation. We performed in vitro experiments using eosinophils, dendritic cells (DCs), and CD4+ T cells obtained from a murine model of asthma. The supernatant of the eosinophil culture was found to significantly increase the levels of interleukin (IL)-4 and IL-5 in the supernatant of CD4+ T cells co-cultured with DCs. HMGB1 levels increased in the supernatant of the eosinophil culture stimulated with IL-5. Anti-HMGB1 antibodies significantly attenuated increases of IL-4 and IL-5 levels in the supernatant of CD4+ T cells co-cultured with DCs that were induced by the supernatant of the eosinophil culture. In addition, anti-HMGB1 antibodies significantly attenuated the expressions of activation markers (CD44 and CD69) on CD4+ T cells. Our data suggest that eosinophils modulate CD4+ T cell responses via HMGB1 in the pathogenesis of asthma.
Antibodies ; Asthma* ; Dendritic Cells ; Eosinophils* ; HMGB1 Protein ; Inflammation ; Interleukin-4 ; Interleukin-5 ; Interleukins ; T-Lymphocytes

BACKGROUND: It is known that immunotherapy promotes the development of allergen-specific Thl-like lymphocytes whose products are effective in inhibiting clinical response of sensitized atopic patients to allergen exposure. At the single cell level in short term culture, however, IL-4 and IL-5 are co-expressed, while IL-4 and IFN-y are exclusively expressed. OBJECTIVE: IL-4, IL-5, and IFN-y mRNA were measured in Der pI-specific T-cell clones (TCCs) to evaluate whether expression of cytokine in allergen-specific TCC is fixed regardless of stimuli. METHOD: Seven Der pI-specific TCCs were made from two asthmatics sensitive to D. pteronyssinus. IL-4, IL-5, and IFN-y mRNA were measured by RT-PCR in these TCCs after antigen-specific (Der pI) and nonspecific (PHA + TPA) stimuli. RESULTS: IL-4 and IL-5 mRNA were expressed in four and six of seven TCCs, but IFN-y mRNA was not expressed in any TCCs after Der pI-specific stimuli. Meanwhile, after the stimulus of TPA plus PHA, IFN-y mRNA as well as IL-4 and IL-5 mRNA were expressed in four of seven TCCs, and in one TCC, only IFN-y mRNA was expressed without expression of IL-4 mRNA. CONCLUSION: The expression of cytokine may be variable in allergen-specific TCC according to the type and amount of stimuli.
Asthma ; Clone Cells* ; Humans ; Immunotherapy ; Interleukin-4* ; Interleukin-5* ; Lymphocytes ; RNA, Messenger* ; T-Lymphocytes*