OBJECTIVETo explore a new experimental method for screening of allergens in post-market traditional Chinese medicine injections by confirming allergens in Huachansu injection.

METHODFirst of all, the serum of patients allergic to Huachansu injection were collected, at the same time, the dubious allergen was conjugated to bovine serum albumin (BSA) by EDC coupling procedure to form complete antigen (BNP-BSA), which makes it possible to reproduce the allergic reaction of Huachansu injection in vitro. The histamine liberation ratio, the level of TNF-alpha and Histamine released from RBL-2H3 mast cell were detected; the above data were compared with those obtained in vivo.

RESULTThe difference of the histamine liberation ratio, the levels of TNF-alpha and histamine of the resibufogenin-BSA group, group of patients allergic to Huachansu injection were not significant compared with those of normal control group. However, there were significant difference in those data among the cinobufagin-BSA group, the blank control and normal control group (P<0.05).

CONCLUSIONThe allergen in the serum collected from patients allergic to Huachansu injection is resibufogenin.

Allergens ; adverse effects ; immunology ; Amphibian Venoms ; adverse effects ; immunology ; Animals ; Anura ; Bufanolides ; adverse effects ; immunology ; Drug Hypersensitivity ; immunology ; Histamine Release ; Humans ; Mast Cells ; immunology ; Medicine, Chinese Traditional

Isocyanate is the most prevalent agent in occupational asthma(OA) in Korea. We analyzed 43 toluene diisocyanate(TDI) induced OA patients of whom 81% were found to be spray painters. The bronchial sensitivity of all subjects was confirmed by TDI-bronchial challenge test. Serum-specific IgE antibodies to isocyanate-human serum albumin(HSA) conjugate were detected by RAST technique(Pharmacia, Sweden). Bronchial challenge test results revealed 21(57%) early, 5 late only, 4 dual, and 12 atypical responders(5 prolonged immediate, 6 square-shaped, 1 progressive). Four(9%) subjects had negative results on the methacholine bronchial challenge test. High levels of serum specific IgE antibody to isocyanate-HSA were found in 17(40%) patients. The prevalence of a specific IgE antibody was not associated with a type of TDI-bronchial challenge test response, smoking and atopic status, presence of rhino-sinusitis and systemic symptoms, or a degree of airway hyperresponsiveness to methacholine(p> 0.05). The period of latency, ranging from 3 to 132 months, was significantly longer in high specific IgE responders (p< 0.05). These data suggest that 40% of isocyanate-induced occupational asthma patients had high specific IgE antibody to isocyanate-HSA conjugate. The presence of specific IgE antibody does not seem to correlate with clinical parameters.
Allergens/adverse effects/*immunology ; Asthma/chemically induced/*immunology ; Female ; Human ; Immunoglobulin E/blood/*immunology ; Male ; Occupational Exposure/*adverse effects ; Toluene 2,4-Diisocyanate/adverse effects/*immunology

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 ; Allergens/analysis+ACo- ; Allergens/adverse effects ; Blotting, Western ; Cells, Cultured ; Comparative Study ; Electrophoresis, Polyacrylamide Gel ; Food Hypersensitivity/immunology ; Heat ; Human ; Hybridization ; Korea ; Lymphocyte Transformation ; North America ; Peanuts/immunology ; Peanuts/classification ; Peanuts/chemistry+ACo- ; Plant Extracts/immunology ; Plant Proteins/immunology ; Plant Proteins/analysis+ACo- ; Plant Proteins/adverse effects ; Skin Tests ; Species Specificity ; T-Lymphocytes/immunology ; T-Lymphocytes/drug effects

Reviewing the progress on study about the major allergen of Shuanghuanglian injection in recent years, resulted in that individual differences of anaphylactic shock are closely related with HLA gene polymorphism. Basing on this, we put forward the research strategy on susceptibility gene of important allergen of Shuanghuanglian injection based on the theory of genetic fingerprints, in order to make sure about the relationship the major allergen of Shuanghuanglian injection and HLA-DRB gene polymorphism and specificity IgE antibody, and to clarify the allergic reaction loci reduced allergic reactions, which can provide the reference data for the study on mechanisms for anaphylactic reaction of Shuanghuanglian injection, and research ideas for the sensitization mechanism of traditional Chinese medicine injection study.
Allergens ; adverse effects ; Anaphylaxis ; chemically induced ; genetics ; immunology ; Drugs, Chinese Herbal ; adverse effects ; Genetic Predisposition to Disease ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Immunoglobulin E ; immunology ; Injections ; Polymorphism, Genetic ; genetics

Allergic disease caused by airborne pollen is a major health problem in China. Intensive study on pollen allergens can be of great help for preventing and treating pollinosis. Four aspects of the study on pollen allergens in China including major allergic pollen in our country, analysis and purification of pollen allergen composition, recombinant pollen allergens and clinical application of pollen allergens are described in this paper.
Air Pollutants ; adverse effects ; Allergens ; administration & dosage ; immunology ; China ; epidemiology ; Desensitization, Immunologic ; methods ; Humans ; Pollen ; immunology ; Recombinant Proteins ; administration & dosage ; immunology ; Rhinitis, Allergic, Seasonal ; epidemiology ; etiology ; prevention & control ; Seasons

Genetically modified (GM) soybean (carrying the EPSPS transgene) is the most common GM food in Korea. In order to assess whether genetic modification increases the allergenic risk of soybeans, the allergenicity and IgE-reactive components of wild-type and GM soybean extracts were compared in allergic adults who had been sensitized to soybeans. We enrolled 1,716 adult allergy patients and 40 healthy, non-atopic controls. Skin prick tests and IgE enzyme linked immunosorbent assays (ELISAs) were performed using wild-type and GM soybean extracts, along with other common inhaled allergens. The specificities of serum IgE antibodies from allergic patients and the identities of the IgE-reactive components of the soybean extracts were compared using ELISA inhibition testing, 2-dimensional gel electrophoresis, and IgE immunoblotting. To evaluate the effects of digestive enzymes and heat treatment, the soybean extracts were heated or pre- incubated with or without simulated gastric and intestinal fluids. The IgE sensitization rates to wild-type and GM soybeans were identical (3.8% of allergic adults), and circulating IgE antibodies specific for the two extracts were comparable. The results of the ELISA inhibition test, SDS-PAGE, and IgE immunoblotting showed a similar composition of IgE-binding components within the wild-type and GM extracts, which was confirmed using two-dimensional gel electrophoresis, IgE immunoblotting, and amino acid sequencing. None of the subjects had a positive response to purified EPSPS protein in the skin prick test, ELISA, or IgE immunoblot analysis. These findings suggest that the IgE sensitization rate to GM soybean extracts is identical to that of wild-type soybean extracts in adult allergy patients. In addition, based on both in vivo and in vitro methods, the allergenicity of wild type and GM soybean extracts was identical.
Soybeans/*immunology ; Skin Tests ; Protein Structure, Tertiary ; *Plants, Genetically Modified ; Middle Aged ; Immunoglobulin E/blood/chemistry ; Immunoblotting ; Humans ; Food Hypersensitivity/etiology/*immunology ; Food/*adverse effects ; Enzyme-Linked Immunosorbent Assay ; Electrophoresis, Gel, Two-Dimensional ; *Crops, Agricultural ; Allergens/*immunology ; Adult ; Adolescent

In Korea, common whelk (Buccinum undatum) is a popular edible shellfish. The aim of this study was to observe the sensitization rate to common whelk and to characterize its allergens. We carried out skin prick test (SPT) in 1,700 patients with various allergic diseases. Specific IgE were detected by ELISA in the patient sera and ELISA inhibition tests were conducted. IgE-binding components were identified by means of SDS-PAGE and IgE-immunoblotting. The effects of diges-tive enzymes were evaluated in both raw and thermally treated extracts. SPT to common whelk was positive (> or =2+) in 83 (4.9%) patients studied. Twenty-four (38.7%) out of 62 SPT positive patients had high serum specific IgE to common whelk. ELISA inhibition test showed significant inhibitions by abalone as well as by common whelk. IgE-immunoblotting demonstrated three IgE-binding components (40, 71, 82 kDa), which were digested by simulated intestinal fluid and moderately digested by simulated gastric fluid, and the digestibility of allergens remained unchanged after thermal treatment. In conclusion, IgE-sensitization rate to com-mon whelk was 4.9% in allergy patients. IgE-immunoblotting demonstrated three IgE-binding components, which were degraded by digestive enzymes. Further studies are needed to evaluate the clinical significance of the sensitized patients to common whelk.
Allergens/immunology ; Animals ; Comparative Study ; Cookery ; Digestion/*physiology ; Food Handling/methods ; Food Hypersensitivity/diagnosis/*epidemiology/*immunology/metabolism ; Heat ; Humans ; Immunoglobulin E/*immunology/metabolism ; Intestines/enzymology ; Korea/epidemiology ; *Mollusca ; Research Support, Non-U.S. Gov't ; Shellfish/*adverse effects ; Skin Tests ; Stomach/enzymology