Identification of IgE-binding components of citrus red mite in sera of patients with citrus red mite–induced asthma

• Published in: Journal of allergy and clinical immunology Vol. 107; no. 2; pp. 244 - 248
• Main Authors: Kim, Hee-Yeon, Park, Hae-Sim, Kim, Yoon-Keun, Son, Jee-Woong, Kim, Hyun-Ah, Suh, Jung-Hee, Nahm, Dong-Ho, Cho, Sang-Heon, Min, Kyung-Up, Kim, You-Young
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.02.2001; Elsevier
• Subjects: Adult; Aged; Allergens - analysis; Allergic diseases; Amino Acid Sequence; amino acid sequencing; Animals; Asthma - blood; Asthma - parasitology; Biological and medical sciences; Citrus - parasitology; Citrus red mite; Electrophoresis, Polyacrylamide Gel - methods; Female; Humans; IgE-binding component; Immunoblotting; Immunoglobulin E - blood; Immunopathology; major allergen; Male; Medical sciences; Middle Aged; Molecular Sequence Data; Panonychus citri; Respiratory and ent allergic diseases; Skin Tests; Sodium Dodecyl Sulfate; Tetranychidae; Trombiculidae - immunology; Tropical medicine; HDM; RT; major allergen; TBST; IgE-binding component; amino acid sequencing; IEF; Citrus red mite; CRM; Human; Immunopathology; Allergy; Tetranychidae; Specific IgE; Respiratory disease; Panonychus citri; Occupational exposure; Asthma; Acariformes; Arachnida; Acari; Actinedida; Arthropoda; Immunoblotting assay; Agriculture; Aeroallergen; Obstructive pulmonary disease; Invertebrata; ELISA assay; Occupational medicine; Aminoacid sequence; Allergen
• ISSN: 0091-6749
• DOI: 10.1067/mai.2001.112123

Background: Our previous investigations demonstrated that citrus red mite (CRM) antigen could cause IgE-mediated bronchoconstriction in exposed farmers working on citrus farms. Objective: This study was performed to confirm IgE-binding components and major allergens within the CRM antigens. Methods: Ten subjects who had been diagnosed as having CRM-induced asthma were enrolled. Serum-specific IgE antibodies to CRM antigens were measured by using an ELISA. To identify IgE-binding components and major allergens, SDS-PAGE, 2-dimensional PAGE, IgE-immunoblot analysis, and amino acid sequencing of major allergens were performed. Results: All the asthmatic subjects had high specific IgE antibodies to CRMs. Twelve percent SDS-PAGE analysis showed more than 10 protein bands ranging from 6 to 64 kd. SDS-PAGE and IgE-immunoblot analysis with each individual serum showed 5 IgE-binding components (11, 24, 35, 40, and 64 kd), with 2 (24 and 35 kd) of them bound in more than 50% of the study subjects. Two-dimensional PAGE and IgE-immunoblot analysis demonstrated that the major allergen at 24 kd had 2 bands with different isoelectric points of 4.75 and 5.1. Thirty-five kilodaltons had one band with an isoelectric point of 4.75. All amino acid sequencing of the 2 major allergens was performed, which was not homologous with any previously characterized allergens. Conclusion: Five IgE-binding components and 2 major allergens (24 and 35 kd) were identified within the CRM antigen. The N-terminal amino acid sequence of the 2 major allergens (24 and 35 kd) was determined. (J Allergy Clin Immunol 2001;107:244-8.)