Dexamethasone (Dex) was shown to inhibit the differentiation, maturation, and antigen-presenting function of dendritic cells (DC) when added during DC generation or maturation stages. Here, we examined the direct effects of Dex on MHC-restricted antigen processing. Macrophages were incubated with microencapsulated ovalbumin (OVA) in the presence of different concentrations of Dex for 2 h, and the efficacy of OVA peptide presentation was evaluated using OVA-specific CD8 and CD4 T cells. Dex inhibited both class I- and class II-restricted presentation of OVA to T cells; this inhibitory effect on antigen presentation was much more potent in immature macrophages than in mature macrophages. The presentation of the exogenously added OVA peptide SIINFEKL was not blocked by Dex. In addition, short-term treatment of macrophages with Dex had no discernible effects on the phagocytic activity, total expression levels of MHC molecules or co-stimulatory molecules. These results demonstrate that Dex inhibits intracellular processing events of phagocytosed antigens in macrophages.
Antigen Presentation* ; Dendritic Cells ; Dexamethasone* ; Macrophages ; Ovalbumin ; Ovum ; T-Lymphocytes

Vaccine contains active components, adjuvants, stabilizers, preservatives, and trace components. Adverse reactions to vaccines are rarely reported. However, all of vaccine components may elicit adverse reaction including life-threatening event in susceptible individuals, therefore raising concerns regarding safety of vaccine still continue up to date. Hypersensitivity reaction to vaccines can be classified as allergic reaction to the vaccine component, pseudo-allergic reaction, and exacerbation of allergic diseases. Hypersensitivity reactions to vaccine components rarely occurred in the population-level, while severe hypersensitivity reaction such as anaphylaxis may be consequent result in susceptible individuals. Some of components such as ovalbumin, gelatin, yeast, and latex were considered as common causes of allergic reactions to the vaccine. However, the incidence or causes of vaccine related hypersensitivity reaction in Korea were not well known. The aims of this article are to review the literatures with hypersensitivity reactions related with vaccine components, to provide detailed information about major components of vaccines commonly used in Korea, and to provide the best way of vaccination in susceptible individuals.
Anaphylaxis ; Gelatin ; Hypersensitivity* ; Incidence ; Korea ; Latex ; Ovalbumin ; Vaccination ; Vaccines ; Yeasts

BACKGROUND: Eosinophilic airway inflammation is a characteristic feature of bronchial asthma. Recent studies showed that increased production and release of eosinophils from bone marrow(BM) might be the essential step in the development of eosinophilic airway inflammation. To testify the hypothesis that increase in BM eosinophil production may be an important determinant of the severity of airway eosinophilia, their relationship was studied in a mouse model of allergic airway inflammation. METHODS: BALB/c mice were sensitized with intraperitoneal ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on two consecutive days. Saline was used for sensitization and challenge in control mice. Bronchoalveolar lavage(BAL) was performed at 24 h after last nasal challenge, immediately followed by BM cell harvest from the femurs. The severity of airway inflammation was assessed as BAL eosinophilia, and the capacity of BM eosinophil production was assessed as BM eosinophil colony forming units(Eo-CFUs) using a semisolid culture assay. RESULTS: There was a significant increase in the percentage of BAL eosinophils and lymphocytes with a resultant decrease in the percentage of alveolar macrophages in the ovalbumin- treated mice, compared with the saline-treated mice(p<0.05, respectively). Change in the percentage of neutrophils was not statistically significant. Compared with the saline-treated mice, the number of BM Eo-CFUs was significantly increased in the ovalbumin- treated mice(p<0.05). But the number of BM Eo-CFUs was not correlated significantly with the number of eosinophils in BAL fluid in the ovalbumin-treated mice. CONCLUSION: Respiratory exposure to allergen induced not only airway eosinophilia but also BM eosinophilopoiesis in this mice model of asthma. However there was no direct relationship between BM eosinophilopoiesis and airway eosinophilia, suggesting that the capacity of eosino-phil production in BM may not an important determinant of severity of airway eosinophilia.
Aluminum ; Animals ; Asthma* ; Bone Marrow* ; Eosinophilia ; Eosinophils* ; Femur ; Inflammation ; Lymphocytes ; Macrophages, Alveolar ; Mice* ; Neutrophils ; Ovalbumin ; Potassium

PURPOSE: Protein cages are promising nanoplatform candidates for efficient delivery systems due to their homogenous size and structure with high biocompatibility and biodegradability. In this study, we investigate the potential of lumazine synthase protein cage as an antigen delivery system to dendritic cells (DCs), which induce antigen-specific T cell proliferation. MATERIALS AND METHODS: Ovalbumin (OVA) peptides OT-1 (SIINFEKL) and OT-2 (ISQAVHAAHAEINEAGR) were genetically inserted to lumazine synthase and each protein cage was over-expressed in Escherichia coli as a soluble protein. The efficiency of antigen delivery and the resulting antigen-specific T cell proliferation by DCs was examined in vitro as well as in vivo. RESULTS: We successfully generated and characterized OVA peptides carrying lumazine synthase protein cages. The OT-1 and OT-2 peptides carried by lumazine synthases were efficiently delivered and processed by DCs in vitro as well as in vivo, and induced proliferation of OT-1-specific CD8+T cells and OT-2-specific CD4+T cells. CONCLUSION: Our data demonstrate the potential of lumazine synthase protein cage being used as a novel antigen delivery system for DC-based vaccine development in future clinical applications.
Antigen Presentation ; Cell Proliferation ; Dendritic Cells ; Escherichia coli ; Nanoparticles* ; Ovalbumin ; Ovum ; Peptides ; Vaccines

BACKGROUND AND OBJECTIVE: The induction of allergic sensitization is the key process of guinea pig asthma model, but few studies has been done on allergen sensitization per se. We performed to investigate the effect of initial sensitization route on asthmatic reactions in response to subsequent repeated allergen challenges. Subjects and METHODS: Thirteen guinea pigs were sensitized with ovalbumin by intraperitoneal injection(group A) and 14 by exposure to aerosolized ovalbumin(group B) on two occasions separated by 1 week. Animals were challenged by inhalation of stepwise increasing concentration of 0.1, 0.5, 1, and 2% ovalbumin every 2 weeks 5 times. We used Penh (enhanced pause) as an index of airway constriction. Allergen inhalation discontinued when an increase of Penh more than 200%, and provocative concentration of ovalbumin causing a 200% increase in Penh was defined as PC200 Bronchoalveolar lavage and histopathological examination of lung tissue was done 24 after last challenge. RESULTS: On first challenge, 11 (85%) of group A had asthmatic reactions, but only 5 (36%) of group B did. Geographic mean of PC200 was significantly lower in group A than that of group B(0.26% vs. 1.26 %, p<0.01). On repeated allergen challenge, PC200 of group A was relatively unchanged, but that of group B was much variable. Sensitized guinea pig had increased eosinophil infiltration in BAL and there are no differences between group A and B in lang pathology. CONCLUSION: Ovalbumin sensitization by intraperitoneal injection of guinea pig is better than inhalation method in first allergen challenge and unvaried asthmatic reactions on repeated challenges.
Animals ; Asthma* ; Bronchoalveolar Lavage ; Constriction ; Eosinophils ; Guinea Pigs* ; Guinea* ; Inhalation* ; Injections, Intraperitoneal* ; Lung ; Ovalbumin* ; Pathology

High-speed counte-recurrent chromatography (HSCCC) is a continuous liquid-liquid partition chromatography without solid matrix, which has the significant features of high resolution and high recovery. The separation of bio-macromolecule in aqueous two-phase systems (ATPs) with HSCCC is still under research, and the establishment of high-speed counter-current aqueous two-phase chromatography (HSCCC-ATP) relies on the improvement of equipment structure and optimization of operation parameters. By using a multi-column high-speed counter-current chromatograph, the separation of protein mixture and the purification of ovalbumin from hen egg white were studied. The effects of pH and PEG concentration on the partition coefficients of proteins were tested in PEG1000-phosphate ATPs, and distinct differences among partition coefficients of proteins were found at pH 9.2 and 15.0% (W/W) PEG concentration in said system. The separation of protein mixture, consisting of cytochrome C, lysozyme and myoglobin was successfully performed in 15.0% (W/W) PEG1000-17.0% (W/W) potassium phosphate ATPs at pH 9.2 with high-speed counter-current chromatograph at rotation speed of 850r/min and flow rate of 0.8mL/min, using upper phase as stationary phase. pH and PEG concentration also had distinct effects on the partition coefficients of the major protein components in hen egg white, including ovaltransferrin, ovalbumin and lysozyme. The optimal pH value and PEG concentration for the purification of ovalbumin by HSCCC-ATP were found to be 9.2 and 16.0% (W/W) respectively. Ovalbumin was successfully purified to homogeneity from the hen egg white sample in 16.0% (W/W) PEG1000-17.0% (W/W) potassium phosphate ATPs at pH 9.2 with high-speed counter-current chromatograph at rotation speed of 850r/min and flow rate of 1.8mL/min, using upper phase as stationary phase. The purification recovery of ovalbumin was around 95%.
Animals ; Chickens ; Countercurrent Distribution ; methods ; Egg White ; chemistry ; Ovalbumin ; isolation & purification

Plasmid vectors with either RSV or CMV promoter are frequently used for DNA- mediated immunization due to the availability in commercial. Consequently, influence of the vector constituents, such as promoter, enhancer and transcription termination signal etc. on vaccination efficiency is not studied extensively. As an initial attempt to develop an efficient vector system for DNA-rnediated immunization, influence of promoter for antigen gene expression on vaccination efficiency has been analyzed. Initially, plasmids with either B-actin or muscle creatine kinase (MCK) promoter were constructed from the plasmid with prototype CMV promoter. In addition, ovalbumin (OVA) antigen gene has been cloned into each vectors to generate the plasmid vectors with different promoters for induction of the anti-OVA immune responses. Antigen protein expression in antigen gene transfected mouse muscle myoblast cells showed that the level from MCK promoter containing plasmid was slightly higher than those from either CMV or B-actin promoter containing plasmids. Also, the same plasmid turned out to be slightly more efficient than other plasmids in antibody imrnune response induction in vivo, when they were applied both through intramuscularly and intradermally. These results suggest that the commonly used CMV promoter containing plasmid vector could be further modified to develop an efficient vector for DNA-mediated immunization.
Animals ; Clone Cells ; Creatine Kinase, MM Form ; Gene Expression ; Immunization ; Mice ; Myoblasts ; Ovalbumin ; Plasmids* ; Vaccination*

OBJECTIVETo establish and evaluate a mouse model of bronchial asthma with Yin deficiency syndrome.

METHODSThe mouse model of bronchial asthma with Yin deficiency syndrome was established by the treatment with injecting ovalbumin (OVA) two times to sensitize, inhaling OVA 14 times to stimulate, and using thyroxin through lavage during late stimulation. This model was evaluated through body weight, asthmatic behaviors, respiratory function, autonomous activity, lung pathology, and pulmonary fluid clearance.

RESULTSOVA combined with thyroxin was an appropriate method to induce the mouse model with increased food and water intake, autonomous activity, asthmatic behaviors score, and respiratory rate, decreased body weight, tidal volume, and wet/dry ratio of lung, and changed with pathology of lung tissue. The changes of the above mentioned parameters indicated that the model was the bronchial asthma with Yin deficiency syndrome.

CONCLUSIONThe OVA combined with thyroxin is a good pattern to establish a mouse model of bronchial asthma with Yin deficiency syndrome successfully, which can highly simulate the clinical symptoms of this disease.

Animals ; Asthma ; physiopathology ; Bronchi ; physiopathology ; Disease Models, Animal ; Mice ; Ovalbumin ; Thyroxine ; Yin Deficiency

Egg allergy is a common problem, especially in young children. In most Westernized countries, milk and egg are the most prevalent foods provoking IgE-mediated symptoms of allergy. Allergens in hen's egg have been well defined and possess immunological characteristics specific to the food. In particular, ovomucoid and ovalbumin, the main allergens, can be partially denaturated by cooking. These aspects have clinical implications as some patients might react only to raw and not to cooked egg. Furthermore, this review will address the specificity of the diagnosis of egg allergy as well as specific clinical problems in egg allergic children.
Allergens ; Child ; Cooking ; Diagnosis ; Egg Hypersensitivity* ; Humans ; Hypersensitivity ; Milk ; Ovalbumin ; Ovomucin ; Ovum* ; Sensitivity and Specificity

BACKGROUND: We reported that level of lipopolysaccharide (LPS) exposure determined the type of airway inflammation in a murine model of asthma. OBJECTIVE: The purpose of this study is to evaluated the role of IL-13 in low dose LPS induced murine model of asthma using IL-13 and signal transducer and activator of transcription 6 (STAT6) deficient mice. METHODS: Mice were sensitized with an intranasal application of LPS-depleted ovalbumin (OA) and different doses of LPS (0.1 and 10 µg), and then challenged intranasally with OA alone. The phenotype changes between wild type (WT) and IL-13-/- mice and between WT and STAT6-/- mice were evaluated. RESULTS: We confirmed again that low and high dose LPS resulted in different phenotypes of murine asthma. In the present study, we observed that phenotypes of murine asthma induced by low dose LPS were abolished in the homozygous null mutation of the IL-13 and STAT6 gene. However, those changes were not shown in mice sensitized OA plus high dose LPS. CONCLUSION: IL-13 plays an important role in low dose LPS induced murine model of asthma. Our results provided a new insight in understanding of the potential role of IL-13 in innate immunity in human allergic asthma.
Animals ; Asthma ; Humans ; Immunity, Innate ; Inflammation ; Interleukin-13 ; Mice ; Models, Animal ; Ovalbumin ; Phenotype ; STAT6 Transcription Factor