PURPOSE: Japanese hop (Humulus spp.) and mugwort (Artemisia spp.) are notable causes of autumn pollinosis in East Asia. However, Japanese hop and mugwort pollen extracts, which are widely used for the diagnosis, have not been standardized. This study was performed to standardize Japanese hop and mugwort pollen extracts. MATERIALS AND METHODS: Allergen extracts were prepared in a standardized way using locally collected Humulus japonicus and purchased Artemisia vulgaris pollens. The immunoglobulin E (IgE) reactivities of prepared extracts were compared with commercial extracts via IgE immunoblotting and inhibition analyses. Intradermal skin tests were performed to determine the bioequivalent allergy unit (BAU). RESULTS: The IgE reactive components of the extracts via IgE immunoblotting were similar to those of commercial extracts. A 11-kDa allergen showed the strongest IgE reactivity in Japanese hop, as did a 28-kDa allergen in mugwort pollen extracts. Allergenic potencies of the investigatory Japanese hop and mugwort extracts were essentially indistinguishable from the commercial ones. Sums of erythema of 50 mm by the intradermal skin test (SigmaED50) were calculated to be 14.4th and 13.6th three-fold dilutions for Japanese hop and mugwort extracts, respectively. Therefore, the allergenic activity of the prepared extracts was 90827.4 BAU/mg for Japanese hop and 34412 BAU/mg for mugwort. CONCLUSION: We produced Japanese hop and mugwort pollen extracts using a standardized method. Standardized Japanese hop and mugwort pollen extracts will facilitate the production of improved diagnostic and immunotherapeutic reagents.
Allergens/*analysis/*immunology ; Antibody Specificity ; *Artemisia ; Bronchial Hyperreactivity/blood/immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoblotting ; Immunoglobulin E/blood/*immunology ; Pollen/*chemistry/*immunology ; Reference Standards ; Republic of Korea ; Rhinitis, Allergic, Seasonal

Wheat dependent exercise-induced anaphylaxis (WDEIA) is a rare but potentially severe food allergy caused by the combination of wheat ingestion and physical exercise. The impact of WDEIA on quality of life (QOL) is unclear. This study characterized the clinical and laboratory features and investigated the QOL in WDEIA patients from Central China. Twenty-eight WDEIA patients were analyzed, and QOL was measured by validated Chinese version Food Allergy Quality of Life Questionnaire-Adult Form (FAQLQ-AF) and Food Allergy Independent Measure (FAIM) after obtaining the diagnosis. The results showed that half of the patients were females. The median onset age was 37 years old. The symptoms occurred within 1 h after wheat ingestion (26/28). Symptoms of anaphylaxis included cutaneous (26/28), respiratory (11/28), gastro-intestinal (5/28) and cardiovascular manifestations (27/28). Skin prick tests were positive to salt soluble (89.3%) and salt insoluble wheat allergen extracts (100%). Positive rate to wheat, gluten and omega-5 gliadin specific IgE was 64.3%, 92.9% and 92.9% respectively. Specific IgE to omega-5 gliadin with a cut-off value 0.83 KU/L offered highly efficient diagnostic criterion for WDEIA (sensitivity: 89.3%; and specificity: 88.9%). The mean scores of FAQLQ-AF and FAIM were 4.70 and 4.98 respectively and level of anti-omega-5 gliadin IgE had positive correlations with FAQLQ scores. Thereby, WDEIA is commonly found in mid-age adults. In most cases, multi-organs especially skin and cardiovascular systems are involved. Salt insoluble wheat allergen skin test and serum specific IgE to gluten and omega-5 gliadin help to diagnose WDEIA. QOL in WDEIA patients is severely impaired.
Adolescent ; Adult ; Aged ; Allergens ; administration & dosage ; chemistry ; immunology ; Anaphylaxis ; diagnosis ; immunology ; physiopathology ; psychology ; China ; Exercise ; Female ; Gastrointestinal Tract ; immunology ; physiopathology ; Gliadin ; administration & dosage ; chemistry ; immunology ; Heart ; physiopathology ; Humans ; Immunoglobulin E ; blood ; Lung ; immunology ; physiopathology ; Male ; Middle Aged ; Quality of Life ; Skin ; immunology ; physiopathology ; Skin Tests ; Surveys and Questionnaires ; Triticum ; chemistry ; immunology ; Wheat Hypersensitivity ; diagnosis ; immunology ; physiopathology ; psychology

Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm.
Adolescent ; Adult ; Allergens/*chemistry/*immunology ; Amino Acid Sequence ; Animals ; Bombyx/*chemistry/genetics/growth & development/*immunology ; Epitopes/immunology ; Female ; Food Hypersensitivity/etiology ; Glycoproteins/*chemistry/genetics/*immunology ; Hot Temperature ; Humans ; Immunoglobulin E/immunology ; Male ; Molecular Sequence Data ; Molecular Weight ; Proteomics ; Pupa/chemistry/immunology ; Recombinant Proteins/biosynthesis/chemistry/immunology ; Sequence Alignment

INTRODUCTIONHealthy individuals may be exposed and sensitised to allergens, and have a positive response to a skin prick test despite being asymptomatic. The objectives of this study were to evaluate the prevalence of atopic sensitisation and identify the reactivity of healthy volunteers to common aeroallergens.

METHODSHealthy volunteers with no known allergic symptoms were recruited in this study. All volunteers were scheduled to undergo a skin prick test with 16 common aeroallergens that were previously identified among atopic patients.

RESULTSA total of 100 volunteers (mean age 28 years) were enrolled in this study. 42 volunteers had positive skin prick tests for at least one allergen. The median number of sensitised allergen was 2 (range 1-7). Volunteers with positive skin tests (n = 42) were younger than those with negative skin tests (n = 58) (mean age 25.5 vs. 29.2 years; p < 0.05). The group with positive skin tests also had a higher proportion of males (57.1% vs. 31.0%; p < 0.01) and first-degree relatives with a history of atopic diseases (31.0% vs. 10.3%; p < 0.05). The most common sensitised allergens in these healthy asymptomatic volunteers were mite (n = 33), house dust (n = 23) and American cockroach (n = 20).

CONCLUSIONIn this study, up to 42% of healthy volunteers, particularly those with a family history of atopy, were sensitised to allergens. Reactivity of the skin test without allergic symptoms, however, does not indicate allergic disease. Therefore, the skin test should only be indicated in atopic symptomatic individuals.

Adult ; Allergens ; chemistry ; Animals ; Cockroaches ; Cross-Sectional Studies ; Dermatitis, Atopic ; diagnosis ; immunology ; Dust ; Female ; Healthy Volunteers ; Humans ; Hypersensitivity ; diagnosis ; immunology ; Male ; Middle Aged ; Mites ; Skin Tests ; methods ; Young Adult

There are some small molecules with potential allergenicity in traditional Chinese medicine injections. They are lack of immunogenicity due to their small molecular weight, but they can lead to allergic reactions when they were coupled with appropriate vectors. Therefore, how to couple small molecule semi-antigens with vectors to prepare complete antigens with immunogenicity and reactogenicity is the key for screening small molecular allergenic substances out of traditional Chinese medicine injections. In terms of semi-antigen characteristics of traditional Chinese medicine injections, vector selection and application, coupling method and complete antigen purification and identification, the author introduces the latest research situations of artificial antigen and antibody preparation technology, the advance in experimental studies on screening of allergenic substances in traditional Chinese medicine injections, as well as the application prospect of immuno-chip technology in studies on allergenic substances in traditional Chinese medicine injections, with the aim of providing new experimental thoughts and methods for safety control of traditional Chinese medicine injections.
Allergens ; administration & dosage ; chemistry ; immunology ; Antigens ; administration & dosage ; chemistry ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Hypersensitivity ; immunology ; Injections ; Medicine, Chinese Traditional ; methods ; trends ; Serum Albumin ; administration & dosage ; chemistry ; immunology

OBJECTIVETo characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn).

METHODSRaw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools.

RESULTSSDS-PAGE of the raw extract showed 23 protein bands (15-250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase.

CONCLUSIONSIt can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.

Allergens ; isolation & purification ; Animals ; Computational Biology ; Electrophoresis, Gel, Two-Dimensional ; Electrophoresis, Polyacrylamide Gel ; Fresh Water ; Humans ; Immunoblotting ; Immunoglobulin E ; immunology ; Mass Spectrometry ; Molecular Weight ; Palaemonidae ; chemistry

Preparation of high quality allergen extracts is essential for the diagnosis and immunotherapy of allergic disorders. Standardization of allergen extracts concerns determination of the allergen unit, development of reference material and measurement of the overall IgE binding capacity of an allergen extract. Recently, quantification of individual allergens has been the main focus of allergen standardization because the allergenicity of most allergen extracts is known to be mainly dependent on the content of a small number of allergen molecules. Therefore, characterization of major allergens will facilitate the standardization of allergens. In this article, we review the current state of allergen standardization. In addition, we briefly summarize the components of allergen extracts that should be under control for the optimization of allergen standardization, since its adjuvant-like activities could play an important role in allergic reactions even though the molecule itself does not bind to the IgE antibodies from subjects.
Adjuvants, Immunologic/chemistry ; Allergens/chemistry/*immunology/isolation & purification ; Polymorphism, Genetic ; Reference Standards ; Republic of Korea

OBJECTIVETo investigate the alteration of inflammasome and receptor during IgG promoter transfected to HepG2 cells.

METHODBy assay of Elisa to evaluate the secretion of IL-1 beta, IL-8, TNF-alpha and MCP-1 after puerarine and LPS administration, and by assay of real time PCR to evaluate the expression of mRNA of IL-1 beta, IL-8,TNF-alpha and MCP-1, as well as the receptors of TLR2, 4 and NOD2, MyD88.

RESULTIgG promoter did not active innate immunity and enhance the expression and secretion of inflammasome in HepG2. Puerarine did not active the inflammasome either. LPS activated the innate immunity and increased the secretion of IL-8, TNF-alpha and MCP-1.

CONCLUSIONIgG-HepG2 cells could be used specifically as the model of allergy type II for ingredients screening. It is suggested that puerarine was suite for the activator for this type of allergy as positive control.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunity, Innate ; immunology ; Immunoglobulin G ; genetics ; Inflammasomes ; immunology ; Promoter Regions, Genetic ; genetics ; Transfection

OBJECTIVETo study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro.

METHODBy transfecting a IgG promoter-regulating the expression of green fluorescent protein(GFP) plasmid into HepG2 cells, this transferred cells were incubated with common allergens (like puerarin, ovalbumin, LPS or Sal typhoid vi polysaccharide vaccine), excipients using in Fufang Kushen injection (NaOH, acetic acid, Tween-80 and ethanol) and samples from the key production processes of the injection for 30 minutes . Fluorescent photographs were analyzed the fluorescence intensity of the cells by using an image analysis software.

RESULTAll of common allergens significantly increased the IgG expression. Two of the excipicents, acetic acids and Tween-80 were shown to increased the IgG expression, while others had no effect on IgG expression. In the 8 samples from the key processes in the production of Fufang Kushen injection, two of them stimulated IgG expression.

CONCLUSIONIgG promoter-HepG2 cells are highly sensitive and specific to allergens, and thus can be applied to rapid screening of allergens in components and injections in transcriptional level. It is possible to use the IgG-promoter HepG2 cells in a real-time monitoring of allergens in the production processes of Chinese medicine injections.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunoglobulin G ; genetics ; Injections ; Medicine, Chinese Traditional ; standards ; Promoter Regions, Genetic ; genetics ; Quality Control

The safety of accelerated schedules of allergen immunotherapy (ASAI) in patients with bronchial asthma (BA) has been reported but there are little data on the safety of ASAI for patients with atopic dermatitis (AD). In this study, we investigated the safety of ASAI in patients with AD. Sixty patients with AD and 18 patients with BA sensitized to house dust mites (HDM) were studied. A maximum maintenance dose of HDM extract, adsorbed to aluminum hydroxide, was administered to patients by subcutaneous injection with either a 3-day protocol (rush immunotherapy) or 1-day protocol (ultra-rush immunotherapy). Systemic reactions were observed 4 of 15 patients (26.7%) with AD during rush immunotherapy, 13 of 45 patients (28.9%) with AD during ultra-rush immunotherapy, and 4 of 18 patients (22.2%) with BA during rush immunotherapy (P > 0.05). No severe or near fatal systemic reactions occurred in 78 subjects of this study. Systemic reactions developed within 4 hr after administration of the maximum allergen dose in 20 of 21 patients (95.2%) with AD and BA who showed systemic reactions during rush or ultra-rush immunotherapy. In conclusion, ASAI was safe and well tolerated in patients with AD. ASAI can be a useful therapeutic option for AD.
Adolescent ; Adult ; Allergens/*therapeutic use ; Aluminum Hydroxide/chemistry ; Animals ; Asthma/therapy ; Dermatitis, Atopic/immunology/*therapy ; Desensitization, Immunologic/*methods ; Drug Administration Schedule ; Female ; Humans ; Infusions, Subcutaneous ; Male ; Pyroglyphidae/*immunology/metabolism