ovalbumin and Leukocytosis

Zizyphus jujuba Mill, a famous oriental traditional medicine, has been reported to exhibit diverse activities in biological systems including the respiratory system. However, a little information is available on its antiasthmatic activity. Jujuboside B (JB) is a natural saponin and one of the active constituent of fruits of Zizyphus jujuba. In the present investigation, JB was isolated from ethanolic extracts of fruits of Zizyphus jujuba (EZJF). EZJF and JB were then evaluated for anti-asthmatic activity using various screening methods. JB was additionally evaluated using ovalbumin (OVA) -induced allergic asthma in mice. Results obtained in the present study showed that EZJF and JB significantly inhibited clonidine-induced catalepsy, milk-induced leucocytosis and eosinophilia, clonidine-induced mast cell degranulation, and passive paw anaphylaxis. The number of inflammatory cells in bronchoalveolar lavage (BAL) fluid was considerably lowered and the severity of pulmonary inflammation was alleviated in the mice pretreated with JB. The high-level expression of T-helper type 2 (TH2) cytokines was markedly reduced in the serum, BAL fluid, and lung homogenates. Thus EZJF and JB showed potent anti-asthmatic activity. Hence EZJF and JB possess a potential role in the treatment of asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Catalepsy; Clonidine; Cytokines; Disease Models, Animal; Eosinophilia; Leukocytosis; Lung; Mast Cells; Medicine, Chinese Traditional; Mice; Milk; Ovalbumin; Plant Extracts; Rats; Rats, Wistar; Saponins; Ziziphus

The mammary gland is able to detect and react to bacterial intrusion through innate immunity mechanisms, but mammary inflammation can also result from antigen-specific adaptive immunity. We postulated that innate and adaptive immune responses could synergize to trigger inflammation in the mammary gland. To test this hypothesis, we immunized cows with the model antigen ovalbumin and challenged the sensitized animals with either Escherichia coli lipopolysaccharide (LPS) as innate immunity agonist, ovalbumin as adaptive immunity agonist, or both agonists in three different udder quarters of lactating cows. There was a significant amplification of the initial milk leukocytosis in the quarters challenged with the two agonists compared to leukocytosis in quarters challenged with LPS or ovalbumin alone. This synergistic response occurred only with the cows that developed the ovalbumin-specific inflammatory response, and there were significant correlations between milk leukocytosis and production of IL-17A and IFN-γ in a whole-blood ovalbumin stimulation assay. The antigen-specific response induced substantial concentrations of IL-17A and IFN-γ in milk contrary to the response to LPS. Such a synergy at the onset of the reaction of the mammary gland suggests that induction of antigen-specific immune response with bacterial antigens could improve the initial immune response to infection, hence reducing the bacterial load and contributing to protection.

Topics: Adaptive Immunity; Animals; Cattle; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Immunity, Innate; Immunization; Inflammation; Interferon-gamma; Interleukin-17; Leukocytosis; Lipopolysaccharides; Mammary Glands, Animal; Mastitis, Bovine; Milk; Ovalbumin; Pilot Projects

Asthma is a chronic inflammatory heterogeneous disorder initiated by a dysregulated immune response which drives disease development in susceptible individuals. Though T helper 2 (TH2) biased responses are usually linked to eosinophilic asthma, other Th cell subsets induce neutrophilic airway inflammation which provokes the most severe asthmatic phenotypes. A growing evidence highlights the role of T regulatory (Treg) cells in damping abnormal Th responses and thus inhibiting allergy and asthma. Therefore, strategies to induce or augment Treg cells hold promise for treatment and prevention of allergic airway inflammation. Recently, the link between Tumor necrosis factor-α (TNF-α) and Treg has been uncovered, and TNF-α antagonists are increasingly used in many autoimmune diseases. Yet, their benefits in allergic airway inflammation is not clarified. We investigated the effect of Adalimumab, a TNF-α antagonist, on Ovalbumin (OVA)-induced allergic airway inflammation in CD1 mice and explored its impact on Treg cells. Our results showed that Adalimumab treatment attenuated the OVA-induced increase in serum IgE, TH2 and TH1 derived inflammatory cytokines (IL-4 and IFN-γ, respectively) in bronchoalveolar lavage (BAL) fluid, suppressed recruitment of inflammatory cells in BAL fluid and lung, and inhibited BAL fluid neutrophilia. It also ameliorated goblet cell metaplasia and bronchial fibrosis. Splenocytes flow cytometry revealed increased percentage of CD4(+) CD25(+) FOXP3(+) Treg cells by Adalimumab that was associated with increase in their suppressive activity as shown by elevated BAL fluid IL-10. We conclude that the beneficial effects of Adalimumab in this CD1 neutrophilic model of allergic airway inflammation are attributed to augmentation of Treg cell number and activity.

Topics: Adalimumab; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Forkhead Transcription Factors; Hyperplasia; Immunoglobulin E; Interleukin-2 Receptor alpha Subunit; Leukocytosis; Lung; Male; Mice; Ovalbumin; Pulmonary Fibrosis; Spleen; T-Lymphocytes, Regulatory

In the present study, we investigated whether the Lagerstroemia indica Linn (LI) extract has an anti-inflammatory effect on lung inflammation in ovalbumin-induced asthmatic mice.. The LI extract was obtained from dried and powdered whole plants of LI using 80% ethanol. ELISA was performed to evaluate cytokine concentration. BALB/c mice were used as a mouse model of asthma after asthmatic induction by ovalbumin sensitization and inhalation. We examined the effects of the LI extract on leukocyte infiltration and mucus secretion using cell count and histological stain.. The amount of cytokines, such as interleukin (IL)-2, IL-4, IL-5, IL-13, and TNF-α, was increased in Jurkat cells using the extract from house dust mites. Increased cytokine concentrations were inhibited by the LI extract. The LI extract suppressed the increased expression of IL-6 after treatment with mite extract of EoL-1 cells and THP-1 cells. In an in vivo experiment using asthmatic mice, the LI extract significantly inhibited leukocytosis and eosinophilia in bronchoalveolar lavage (BAL) fluid and lung tissue samples. The LI extract inhibited the increase in mucus secretion by goblet cells, blocked the production of reactive oxygen species in BAL fluid cells, and blocked the protein expression of IL-5 in BAL fluid. The concentration of ovalbumin-specific IgE in BAL fluid was weakly inhibited by the LI extract.. These results suggest that the LI extract may be used as a valuable agent for treating allergic diseases such as asthma due to its anti-inflammatory property.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Eosinophilia; Eosinophils; Female; Goblet Cells; Humans; Immunoglobulin E; Inflammation; Lagerstroemia; Leukocytosis; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Phytotherapy; Plant Extracts; Pyroglyphidae; Reactive Oxygen Species

Duchesnea chrysantha (D. chrysantha) is a herb with anti-oxidative, anti-inflammatory and immune-enhancing properties.. Asthma is an inflammatory disease of the lungs, and the hallmarks of the disease are increased inflammatory cell infiltration into the airways and poor respiratory function. Although there is the possibility that D. chrysantha may have an inhibitory effect on lung inflammation, the effects of D. chrysantha on asthma have not been fully investigated. In the present study, we investigated the anti-inflammatory activity of D. chrysantha extract (Dc extract) on lung inflammation in a murine model of ovalbumin-induced asthma.. Dc extract was obtained from dried and powdered whole plants of D. chrysantha using 80% ethanol. BALB/c mice induced by ovalbumin sensitization and nebulization were used as a mouse model of asthma. RT-PCR and ELISA were performed to measure mRNA and protein expression of cytokines. We examined the effects of Dc extract on leukocyte infiltration and mucus secretion using periodic acid-Schiff staining as well as hematoxylin and eosin staining.. Dc extract significantly inhibited leukocytosis and eosinophilia in the bronchoalveolar lavage (BAL) fluid (p<0.01). Dc extract significantly reduced the elevated infiltration of inflammatory cells (p<0.05) and inhibited the increased mucus secretion, despite the absence of significant value. Although Dc extract weakly inhibited the mRNA expression of IL-4, IL-5, IL-13, and eotaxin, it strongly inhibited the protein expression of IL-5 (p<0.05) and eotaxin (p<0.01) in BAL fluid. Ovalbumin-specific IgE levels in the serum and BAL fluid were blocked by Dc extract (p<0.05).. These results suggest the possibility that Dc extract can exert suppressive effects on asthma and may provide evidence that Dc extract is a useful agent for the treatment of allergic airway disease.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Eosinophilia; Female; Immunoglobulin E; Inflammation; Leukocytosis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Reverse Transcriptase Polymerase Chain Reaction; Rosaceae

We have demonstrated previously that local, adenoviral-mediated gene transfer of vIL-10 to a single joint of rabbits and mice with experimental arthritis can suppress disease in both the treated and untreated contralateral joints. These therapeutic effects observed in distant untreated joints following local intra-articular gene delivery have been termed the 'contralateral effect'. To begin to understand the underlying immunologic mechanism that confers this effect, a dual-antigen model of antigen-induced arthritis (AIA) in rabbit knee joints was utilized. Rabbits were immunized against two antigens, ovalbumin and keyhole limpet hemocyanin, and AIA generated by intra-articular injection of each antigen into contralateral knees. Intra-articular adenovirus-mediated gene transfer of vIL-10 significantly reduced intra-articular leukocytosis and cartilage matrix degradation, while preserving near normal levels of cartilage matrix synthesis within treated joints. However, no antiarthritic effect was conferred in the contralateral control joints that received only a marker gene, in contrast to the results seen in a single-antigen AIA model. These results suggest that the distant antiarthritic effects associated with local gene delivery to joints are antigen-specific, and not due to vIL-10-induced generalized immunosuppression of the animal.

Topics: Adenoviridae; Animals; Arthritis, Experimental; Autoimmunity; Cartilage, Articular; Disease Progression; Female; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors; Hemocyanins; Injections, Intra-Articular; Interleukin-10; Leukocytosis; Ovalbumin; Rabbits; Synovial Membrane

Ag-specific Th1 and Th2 cells have been demonstrated to play a critical role in the induction of allergic diseases. Here we have investigated the precise mechanisms of Th1-induced airway inflammation. Airway inflammation was induced in BALB/c mice by transfer of freshly induced OVA-specific Th1 or Th2 cells followed by OVA inhalation. In this model, both Th1 and Th2 cells induced airway inflammation. The former induced neutrophilia in airways, whereas the latter induced eosinophilia. Moreover, we found that Th1 cells induced more severe airway hyperresponsiveness (AHR) than Th2 cells. The eosinophilia induced by Th2 cell infusion was almost completely blocked by administration of anti-IL-5 mAb, but not anti-IL-4 mAb. In contrast, Th1-induced AHR and pulmonary neutrophilia were inhibited by the administration of anti-human IL-8R Ab, which blocks the function of mouse CXC chemokine(s). These findings reveal a critical role of mouse CXC chemokine(s) in Th1-dependent pulmonary neutrophilia and AHR.

Topics: Administration, Inhalation; Aerosols; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemokines, CXC; Disease Models, Animal; Eosinophils; Inflammation; Leukocytosis; Lung; Mice; Mice, Inbred BALB C; Mice, Transgenic; Neutrophils; Ovalbumin; Peptide Fragments; Respiratory Hypersensitivity; Th1 Cells; Th2 Cells

The involvement of the sympathetic and dopaminergic systems on blood neutrophilic leucocytosis observed during anaphylaxis was investigated. Blood neutrophil counts impressively increased 1 h after intravenous injection of ovalbumin (OVA, 250 micrograms/kg) into OVA-immunized rats. The increase in neutrophil counts induced by OVA was abrogated after catecholamine depletion by reserpine. Either adrenalectomy or the alpha- and beta-adrenoceptor antagonists phentolamine and propranolol, respectively, had only minor inhibitory effects on neutrophilia induced by antigen. On the other hand, pretreatment with the dopaminergic antagonists chlorpromazine and pimozide significantly inhibited the neutrophilia. The intravenous injection of apomorphine, a dopaminergic agonist, increased neutrophil counts in naive animals, while chlorpromazine completely inhibited this phenomenon. These results suggest that dopaminergic mechanisms play a role in the systemic neutrophilia observed during anaphylactic shock.

Topics: Adrenergic Antagonists; Anaphylaxis; Animals; Dopamine Antagonists; Leukocytosis; Male; Neutrophils; Ovalbumin; Rats; Rats, Wistar; Reserpine

Pertussis toxin is produced by the causative agent of whooping cough, Bordetella pertussis, and is an adenosine diphosphate (ADP)-ribosyltransferase capable of covalently modifying and thereby inactivating many eukaryotic G proteins involved in cellular metabolism. The toxin is a principal determinant of virulence in whooping cough and is a primary candidate for an acellular pertussis vaccine, yet it is unclear whether the ADP-ribosyltransferase activity is required for both pathogenic and immunoprotective activities. A B. pertussis strain that produced an assembled pertussis holotoxin with only 1 percent of the ADP-ribosyltransferase activity of the native toxin was constructed and was found to be deficient in pathogenic activities associated with B. pertussis including induction of leukocytosis, potentiation of anaphylaxis, and stimulation of histamine sensitivity. Moreover, this mutant strain failed to function as an adjuvant and was less effective in protecting mice from intracerebral challenge infection. These data suggest that the ADP-ribosyltransferase activity is necessary for both pathogenicity and optimum immunoprotection. These findings bear directly on the design of a nontoxic pertussis vaccine.

Topics: Adjuvants, Immunologic; ADP Ribose Transferases; Anaphylaxis; Animals; Antigens; Bordetella pertussis; Codon; Drug Tolerance; Histamine; Immunization; Leukocytosis; Macromolecular Substances; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mutation; Ovalbumin; Pentosyltransferases; Pertussis Toxin; Virulence Factors, Bordetella

A highly potent extract of the histamine sensitizing factor (HSF) of Bordetella pertussis was isolated by extraction of bacterial cells with urea buffer and subsequent gel filtration. This preparation of HSF also contained leukocytosis-promoting activity and adjuvant activity for reaginic and hemagglutinating antibodiesl Digestion of this extract with pronase or trypsin partially destroyed histamine-sensitizing activity, leukocytosis-promoting activity, and adjuvant activity for reaginic antibody, but did not affect adjuvant activity for hemagglutinating antibody. Antisera to HSF was prepared by immunizing rabbits with either whole bacteria or partially purfied extract. These antisera contained several precipitating antibodies to Bordetella pertussis extract demonstrated by immunodiffusion and immunoelectrophoresis. Antisera added in vitro to Bordetella pertussis extracts or passively administered in vivo to mice, reduced or abolished all biologic activities except adjuvant activity for hemagglutinating antibody. These results suggest that HSF might be an antigenic component of Bordetella pertussis which also possesses leukocytosis-promoting activity and adjuvant activity for reaginic antibody.

Topics: Adjuvants, Immunologic; Amino Acids; Animals; Biological Assay; Bordetella pertussis; Deoxyribonucleases; Erythrocytes; Female; Hemagglutination Tests; Histamine; Histamine Release; Hypersensitivity, Immediate; Immune Sera; Immunization, Passive; Immunodiffusion; Injections, Intraperitoneal; Leukocytosis; Mice; Ovalbumin; Passive Cutaneous Anaphylaxis; Pronase; Reagins; Ribonucleases; Trypsin

Topics: Anaphylaxis; Animals; Antigen-Antibody Reactions; Capillary Permeability; Carbon; Chemotaxis; Complement System Proteins; Eosinophilia; Fluorescent Antibody Technique; Guinea Pigs; Hypersensitivity, Immediate; Immune Sera; Immunoglobulin E; Immunoglobulin G; Inflammation; Leukocytosis; Mast Cells; Neutrophils; Ovalbumin; Phagocytosis; Proteins; Skin

Topics: Anaphylaxis; Animals; Anterior Chamber; Capillary Permeability; Guinea Pigs; Leukocytosis; Ovalbumin; Uvea; Vitreous Body