1.Ovalbumin fused with diphtheria toxin protects mice from ovalbumin induced anaphylactic shock.
Bong Ki LEE ; Young Gun YOO ; Won Young LEE ; Chun Soo HONG ; Jae Ku PARK ; Jai Youl RO
Yonsei Medical Journal 2001;42(1):91-105
For those with allergy, vaccination with a specific allergen has often been used as a major therapeutic measure. However, the universal application of this technique in clinics have been restricted due to its low success rates and the risk of active systemic anaphylactic shock (ASAS). In this regard, we constructed a fusion protein (OVA-DT), ovalbumin (OVA) fused with diphtheria toxin protein (DT), which may exert a specific cytotoxicity to cells bearing OVA-specific IgE. Its therapeutic effect was evaluated in mice (BALB/c) sensitized with OVA (Os-mice). OVA challenges to the OVA-sensitized mice (Os-mice) caused ASAS to death within 30 min, but OVA-DT treatment afforded mice complete protection. When OVA-DT was treated to the Os-mice, none showed the signs of ASAS when re-challenged 48 h after the treatment. OVA-DT itself was not found to be toxic or allergenic in normal mice. The effect of OVA-DT on the biological functions of mast cells was also studied. Binding of OVA-DT to OVA-specific IgE bearing mast cells and the inhibition of histamine release from these cells were observed. In addition, OVA-DT treatment inhibited the proliferation of OVA-specific B cells in mice. In Os-mice treated with OVA-DT, levels of anti-OVA IgG2a in serum and the production of IFN-gamma by splenic lymphocytes were found to increase, but the production of IL-4 by these cells decreased. Re-direction of cytokine profiles from OVA-specific Th2 to OVA-specific Thl is suggested. These results indicate that OVA-DT can protect Os-mice from ASAS due to OVA challenge, because it inactivates OVA-specific IgE-expressing cells, including mast cells and B cells.
Anaphylaxis/prevention | control*
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Animal
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B-Lymphocytes/immunology
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Female
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Histamine Release/drug effects
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IgE/metabolism
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Interferon Type II/biosynthesis
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Interleukin-4/biosynthesis
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Lymphocyte Transformation/drug effects
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Mast Cells/metabolism
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Mice
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Mice, Inbred BALB C
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Ovalbumin/immunology*
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Recombinant Fusion Proteins/therapeutic use*
2.A comparison study on allergen components between Korean (Arachis fastigiata Shinpung) and American peanut (Arachis hypogaea Runner).
Chun Wook PARK ; Gyeong Il KIM ; Cheol Heon LEE
Journal of Korean Medical Science 2000;15(4):387-392
The prevalence of peanut allergy in Korea is lower than in America. Peanut extract allergens between the two countries have not been standardized. This study was performed to compare the allergenicity of raw Korean and American peanuts with that of roasted peanuts. We prepared the peanut extracts in Korean raw (KP) and roasted peanuts (KRP), and also in American raw (AP) and roasted (ARP) peanuts. We compared the peanut extract allergens of KP, KRP, AP and ARP in vitro with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting, T-cell proliferation assay and skin prick test with sera from peanut-allergic patients. SDS-PAGE and Western blotting demonstrated four allergenic extracts, numerous bands that displayed a high prevalence of IgE binding. IgE-binding bands were at 64, 36 and 17 kDa. Western blot inhibition revealed that either KP or AP could almost completely inhibit the reactivity of the other extract. There were no differences between T-cell proliferation assay and skin prick test. In conclusion, this investigation showed no different allergic components in both raw and roast extracts of Korean and American peanuts.
Allergens/immunology
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Allergens/analysis+ACo-
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Allergens/adverse effects
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Blotting, Western
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Cells, Cultured
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Comparative Study
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Electrophoresis, Polyacrylamide Gel
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Food Hypersensitivity/immunology
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Heat
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Human
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Hybridization
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Korea
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Lymphocyte Transformation
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North America
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Peanuts/immunology
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Peanuts/classification
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Peanuts/chemistry+ACo-
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Plant Extracts/immunology
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Plant Proteins/immunology
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Plant Proteins/analysis+ACo-
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Plant Proteins/adverse effects
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Skin Tests
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Species Specificity
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T-Lymphocytes/immunology
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T-Lymphocytes/drug effects