Bacille Calmette-Guerin (BCG) induces potent Th1 responses with the help of interleukin (IL)-10 and IL-12 released from dendritic cells (DCs), and suppresses Th2- associated allergic reactions. However, there are still some controversies on therapeutic effects of BCG in asthmatics. This study investigated whether BCG administration to DCs suppresses IL-5 production from T cells in atopic asthmatics. DCs derived from peripheral blood of subjects were cultured with or without BCG and Dermatophagoides farinae extract. Some DCs were co-cultured with T cells in the presence of BCG or the above culture supernatants. In the atopic asthmatics, BCG significantly increased IL-10 and IL-12 production from DCs. In the presence of D. farinae extract, BCG further increased IL-10 production. BCG-induced IL-10 production was significantly higher in the atopics (n=14) than in the non-atopics (n=9). Both BCG and the BCG-treated DCs culture supernatant significantly increased IFN-gamma production from T cells. Both BCG and the supernatant from DCs+BCG+D. farinae co-cultures significantly decreased IL-5 production (all p<0.05), but the supernatant from DCs+BCG co-cultures did not. In conclusion, administration of BCG together with D. farinae extract effectively decreased IL-5 production from T cells, probably through the action of IL-10 and IL-12 released from DCs in D. farinaesensitive asthmatics.
Adult ; Asthma/*immunology ; BCG Vaccine/*immunology ; Dendritic Cells/*immunology ; Female ; Humans ; Interferon-gamma/biosynthesis ; Interleukin-10/biosynthesis ; Interleukin-12/biosynthesis ; Interleukin-5/*biosynthesis ; Lymphocyte Activation ; Male ; T-Lymphocytes/*immunology

OBJECTIVETo observe the inhibitory effect of mizolastine on substance P(SP)-induced production of leukotriene B(4) (LTB(4)) and interleukin 5 (IL-5) in mouse skin.

METHODSMice were fed with different doses of mizolastine or other control drugs, 30 min after administration animals were injected intradermally with SP on the back. The treated skin samples were taken and competitive enzyme-link immunoassay (ELISA) method was applied to detect LTB (4) and IL-5 in the skin samples.

RESULTThe LTB(4) and IL-5 levels in 10 mg/kg mizolastine group were (1.23 +/-0.29)pg/ml and (34.28 +/-11.00)pg/ml, respectively, which were lower than those in saline control group [(5.52+/-1.88)pg/ml and (179.62 +/-46.25)pg/ml respectively] or loratadine group [(3.89+/-1.27)pg/ml and (127.74 +/-43.27)pg/ml respectively]. No significant difference was found between 10 mg/kg mizolastine group and dexamethasone group (P=0.161 and P=0.508).

CONCLUSIONMizolastine might inhibit the production of LTB(4) and IL-5 induced by substance P in mouse skin, suggesting that anti-inflammatory effect and the blockade of histamine H1 receptors might be involved in its anti-pruritic mechanisms.

Animals ; Benzimidazoles ; pharmacology ; Female ; Histamine H1 Antagonists ; pharmacology ; Interleukin-5 ; biosynthesis ; Leukotriene B4 ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; Skin ; metabolism ; Substance P ; antagonists & inhibitors

In order to get a better understanding of the role of protease-activated receptor 2 (PAR2) in type 2 helper T (Th2) cell responses against Trichinella spiralis infection, we analyzed Th2 responses in T. spiralis-infected PAR2 knockout (KO) mice. The levels of the Th2 cell-secreted cytokines, IL-4, IL-5, and IL-13 were markedly reduced in the PAR2 KO mice as compared to the wild type mice following infection with T. spiralis. The serum levels of parasite-specific IgE increased significantly in the wild type mice as the result of T. spiralis infection, but this level was not significantly increased in PAR2 KO mice. The expression level of thymic stromal lymphopoietin, IL-25, and eotaxin gene (the genes were recently known as Th2 response initiators) of mouse intestinal epithelial cells were increased as the result of treatment with T. spiralis excretory-secretory proteins. However, the expression of these chemokine genes was inhibited by protease inhibitor treatments. In conclusion, PAR2 might involve in Th2 responses against T. spiralis infection.
Animals ; Antibodies, Helminth/blood ; Chemokine CCL11/biosynthesis ; Cytokines/biosynthesis ; Female ; Gene Expression Profiling ; Immunoglobulin E/blood ; Interleukin-13/secretion ; Interleukin-4/secretion ; Interleukin-5/secretion ; Interleukins/biosynthesis ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Receptor, PAR-2/*metabolism ; Th2 Cells/*immunology ; Trichinella spiralis/*immunology ; Trichinellosis/*immunology

OBJECTIVETo observe the effect of three-stage sequential method (TSSM) on interleukin-5 (IL-5) mRNA expression in the lung tissue of asthmatic rat model treated with steroid.

METHODSSD rats were randomly divided into 8 groups, the normal control group (GA), the model group (GB), the model with steroid intervention group (GC) and the 5 treated groups (GD - GH). Excepting those in the GA, all rats were established into the asthma model and rats in GC-GH were intervened by dexamethasone, with the dosage reduced by 0.1 mg/kg per week starting from the 3rd week and withdrawn completely till the 7th week. At the same time, rats in GD - GH were treated with TSSM, and the 1st, 2nd and 3rd recipe of TSSM, as well as pulmicort respules (as positive control) respectively. IL-5 mRNA expression in the lung tissue of rats was detected at different time points by in situ hybridization.

RESULTSComparisons of GD with other groups in IL-5 mRNA expression showed: it was remarkably lowered than that in GB and GC at all time points (P < 0.01); compared to GH, the difference was insignificant at the end of the 2nd week, but significant at the end of the 7th and 9th week (P < 0.01); as for the difference between GD with GE, GF and GG, it showed very significant difference (P < 0.01) at all the time points besides that at the end of the 2nd week, it showed insignificance to GE (P > 0.05) and significance to GF (P < 0.05). The dynamic changes of IL-5 mRNA expression in GD during the steroid withdrawal period showed the lowering was more significantly at the end of the 7th and 9th week than that at the end of the 2nd week (P < 0.05, P < 0.01), but its levels were equal at the former two time points; its lowest level appeared at the end of the 9th week, which approaching the level in GA (P > 0.05).

CONCLUSIONThree-stage sequential method could remarkably inhibit IL-5 mRNA expression in the lung tissue of asthmatic rats undergoing steroid treatment.

Animals ; Asthma ; drug therapy ; Dexamethasone ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Interleukin-5 ; biosynthesis ; genetics ; Lung ; metabolism ; Male ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the effect of intrapulmonary regulatory peptides on adhesion of eosinophils (EOS) to bronchial epithelial cells (BECs). METHODS: Two regulatory peptides, namely vasoactive intestinal peptide (VIP) and epidermal growth factor (EGF) were investigated. VIP and EGF were observed on the secretion of ILs and expression of intercellular adhesion molecule-1 (ICAM-1). RESULTS: VIP and EGF could decrease ILs secretion and ICAM-1 expression. CONCLUSION VIP and EGF inhibited the adhesion of EOS to BEC in the inflammatory process to lighten the airway inflammation.
Animals ; Bronchi ; cytology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Eosinophils ; cytology ; Epidermal Growth Factor ; physiology ; Epithelial Cells ; cytology ; Female ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Interleukin-5 ; biosynthesis ; Male ; Rabbits ; Vasoactive Intestinal Peptide ; physiology

OBJECTIVETo explore the regularity of recipe composition by observing inhibitory effects on the genic expression of 5-lipoxygenase activating protein, IL-4 and the leukotriene C4 in asthmatic mice.

METHODThe mice were challenged with OVA and administered ig with the Herba Ephedrae decoction (HED), separated compositions (2500 mg x kg(-1), calculated by Herba Ephedrae) and dexamethasone (2 mg x kg(-1)) respectively once daily for seven days. The real-time fluorescence quantitative PCR method was employed to measure the contents of FLAP mRNA and IL-4 mRNA expressions in lung and the ELISA method was used to determine the content of LTC4 in the washing solution of pulmonary alveolus and bronchi.

RESULTIn the lung of asthma mice, the expressions of FLAP and IL-4 and the content of LTC4 were significantly augmented compared with the control group. The HED and the separated compositions could suppress the expressions of FLAP and IL-4 and LTC4 release to a great extent in mice.

CONCLUSIONThe HED had the remarkable effects of antianaphylaxis asthma and the original formula HED worked best. These results confirmed the rationality and scientific level of HED.

5-Lipoxygenase-Activating Proteins ; Animals ; Asthma ; chemically induced ; genetics ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Carrier Proteins ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Ephedra sinica ; chemistry ; Interleukin-4 ; biosynthesis ; genetics ; Leukotriene C4 ; metabolism ; Lung ; drug effects ; metabolism ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Mice ; Ovalbumin ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation

BACKGROUNDEosinophils are highly related to allergic asthma inflammation. Interleukin (IL)-5 is the major chemokine of eosinophils, inhibition of the activity of IL-5 thus seems to be a potential approach to asthma therapy. The current study was performed to determine whether a recombinant human IL-5 protein as a xenogeneic vaccine has the capability of inducing anti-asthma activities.

METHODSRecombinant human IL-5 was used as a protein vaccine. Mouse asthma model was established to observe the anti-asthma activities. Lung histology was observed; eosinophils in blood and bronchoalveolar lavage were stained and counted. Airway hyperresponsiveness was determined by whole body plethysmograph. Antibody characters and cytokines were detected with enzyme linked immunosorbent assay (ELISA) and Western blot assay.

RESULTSVaccination with recombinant human IL-5 protein as vaccine significantly reduced airway inflammation and airway hyperresponsiveness, and shifted the cytokine production from Th2 (IL-4) to Th1 (INF-gamma) in mice allergic-asthma model. Immunization with recombinant human IL-5 protein vaccine bypassed the immunological tolerance and induced production of polyclonal antibodies that were cross-reactive with murine IL-5.

CONCLUSIONSActive immunization with xenogeneic homologous IL-5 may be a possible therapeutic approach to the treatment of asthma and potentially of other eosinophilic disorders.

Animals ; Asthma ; therapy ; B-Lymphocytes ; immunology ; Bronchial Hyperreactivity ; prevention & control ; Cytokines ; biosynthesis ; Disease Models, Animal ; Interleukin-5 ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; immunology ; Th2 Cells ; immunology ; Vaccination

BACKGROUNDCorticosteroids remain the most effective therapy available for asthma. They have widespread effects on asthmatic airway inflammation. However, little is known about the effects of corticosteroids on the production of bone marrow inflammatory cells in asthma. This study observed the effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells, so as to explore the possible effectiveness of a bone marrow-targeted anti-inflammatory strategy.

METHODSBalb/c mice were sensitized and challenged with ovalbumin (OVA) to establish an asthmatic model. For two consecutive weeks, asthmatic mice were challenged with OVA while being given either prednisone, montelukast, prednisone plus montelukast, or sterile saline solution. The mice were killed 24 hours after the last challenge with OVA, and bronchoalveolar lavage fluid (BALF), peripheral blood, and bone marrow were collected. Eosinophils in peripheral blood and BALF, and nucleated cells in BALF, peripheral blood, and bone marrow were counted. The percentages of CD34+ cells, CD4+ T lymphocytes and CD8+ T lymphocytes among nucleated cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect expression of CD34 and interleukin (IL)-5Ralpha mRNA (CD34+ IL-5Ralpha mRNA+ cells) among bone marrow hematopoietic cells.

RESULTSCompared with the sterile saline solution group, the number of eosinophils in BALF and peripheral blood, CD34+ cells in peripheral blood and bone marrow, and CD34+ IL-5Ralpha mRNA+ cells in bone marrow of mice from the prednisone and prednisone plus montelukast groups were significantly lower (P < 0.01). The number of eosinophils in BALF from the montelukast group was also significantly lower (P < 0.05).

CONCLUSIONSThe results suggest that, in this asthmatic mouse model, prednisone probably inhibits proliferation, differentiation, and migration of CD34+ cells in bone marrow, blocks eosinophilopoiesis in bone marrow, and interferes with eosinophil migration into peripheral blood and subsequent recruitment in the airway. In addition, montelukast may suppress eosinophil infiltration into the lungs of asthmatic mice. However, a significant inhibitory effect of montelukast on the proliferation and migration of CD34+ cells and a cooperating effect with prednisone on bone marrow of asthmatic mice were not observed.

Acetates ; pharmacology ; Animals ; Antigens, CD34 ; analysis ; Asthma ; drug therapy ; Cell Count ; Hematopoietic Stem Cells ; drug effects ; Immunohistochemistry ; In Situ Hybridization ; Interleukin-5 ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; Prednisone ; pharmacology ; Quinolines ; pharmacology

Food allergies affect about 4% of the Korean population, and buckwheat allergy is one of the most severe food allergies in Korea. The purpose of the present study was to develop a murine model of IgE-mediated buckwheat hypersensitivity induced by intragastric sensitization. Young female C3H/HeJ mice were sensitized and challenged intragastricly with fresh buckwheat flour (1, 5, 25 mg/dose of proteins) mixed in cholera toxin, followed by intragastric challenge. Anaphylactic reactions, antigen-specific antibodies, splenocytes proliferation assays and cytokine productions were evaluated. Oral buckwheat challenges of sensitized mice provoked anaphylactic reactions such as severe scratch, perioral/periorbital swellings, or decreased activity. Reactions were associated with elevated levels of buckwheatspecific IgE antibodies. Splenocytes from buckwheat allergic mice exhibited significantly greater proliferative responses to buckwheat than non-allergic mice. Buckwheat-stimulated IL-4, IL-5, and INF-gamma productions were associated with elevated levels of buckwheat-specific IgE in sensitized mice. In this model, 1 mg and 5 mg dose of sensitization produced almost the same degree of Th2-directed immune response, however, a 25 mg dose showed blunted antibody responses. In conclusion, we developed IgE-mediated buckwheat allergy by intragastric sensitization and challenge, and this model could provide a good tool for future studies.
Anaphylaxis/blood/immunology ; Animals ; Cell Proliferation/drug effects ; Comparative Study ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Fagopyrum/*immunology ; Female ; *Flour ; Food Hypersensitivity/blood/*immunology ; Immunoglobulin E/blood/immunology ; Immunoglobulin G/blood/immunology ; Interferon Type II/biosynthesis ; Interleukin-4/biosynthesis ; Interleukin-5/biosynthesis ; Mice ; Mice, Inbred C3H ; Plant Extracts/administration & dosage/immunology ; Research Support, Non-U.S. Gov't ; Spleen/cytology/drug effects/metabolism ; Stomach/drug effects/*immunology ; T-Lymphocytes/cytology/drug effects/metabolism ; Time Factors

Several myeloid leukemia-derived cells have been reported to possess the ability to differentiate into dendritic cells (DC). MUTZ-3, a myeloid leukemia cell line, responds to GM-CSF, IL-4 and TNF-alpha, and acquires a phenotype similar to immature monocyte-derived DC (MoDC). In the present study, MUTZ-3-derived DC (MuDC) showed high level expression of HLA class II molecules, CD80 and CD86, and were able to function as potent antigen presenting cells as previously reported. Interestingly, MuDC maturation was induced by CD40-mediated stimulation, but not by LPS stimulation. We analyzed CCR1, CCR7 and Toll-like receptor (TLR) expressions in MuDC, and measured IL-10 and IL-12 production after maturation stimuli. Although MuDC expressed the mRNA for TLR4, a major component of the LPS receptor system, they did not show an enhanced level of CCR7 or cytokine production after LPS stimulation. In contrast, they responded to CD40 stimulation, which resulted in increased levels of CD83, CD86 and CCR7. Moreover, while LPSstimulated MoDC could potently stimulate NK cells in a DC-NK cell co-culture, LPS-stimulated MuDC failed to stimulate primary NK cells. Taken together, our findings suggest that, although MuDC express TLR4, unlike TNF-alpha and IL-1beta, LPS does not stimulate MuDC to acquire mature phenotypes, and they may have impaired activity to initiate innate immune response.
Antigens, CD40/metabolism/pharmacology ; Antigens, CD80/metabolism ; Antigens, CD86/metabolism ; Blotting, Western ; CD40 Ligand/metabolism/pharmacology ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Dendritic Cells/*drug effects/metabolism ; Enzyme-Linked Immunosorbent Assay ; Fluorescein-5-isothiocyanate ; Fluorescent Antibody Technique, Indirect ; Fluorescent Dyes ; Humans ; Interleukin-10/analysis/biosynthesis ; Interleukin-12/analysis/biosynthesis ; Killer Cells, Natural/metabolism ; Leukemia, Myeloid/*pathology ; Lipopolysaccharides/*pharmacology ; Mitogen-Activated Protein Kinase 3/metabolism ; RNA, Messenger/metabolism ; Research Support, Non-U.S. Gov't ; Reverse Transcriptase Polymerase Chain Reaction ; Toll-Like Receptor 4/metabolism ; Tumor Necrosis Factor-alpha/pharmacology ; p38 Mitogen-Activated Protein Kinases/metabolism