A new method for histamine release from purified peripheral blood basophils using monoclonal antibody-coated magnetic beads

• Published in: Journal of immunological methods Vol. 240; no. 1; pp. 39 - 46
• Main Authors: Nishi, Hiroshi, Nishimura, Shinji, Higashiura, Masahito, Ikeya, Norie, Ohta, Hideki, Tsuji, Tetsuo, Nishimura, Masaji, Ohnishi, Shigeo, Higashi, Hideo
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 23.06.2000; Elsevier
• Subjects: Allergy testing; Antibodies, Monoclonal; Antibody Specificity; Basophil; Basophils - cytology; Basophils - immunology; Biological and medical sciences; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Histamine - analysis; Histamine release; Histamine Release - immunology; Humans; Hypersensitivity - diagnosis; Immediate hypersensitivity. Allergy. Anaphylaxis, etc; Immunobiology; Immunomagnetic Separation - methods; Magnetic beads; magnetism; Positive selection; Techniques; DP, Dermatophagoides pteronyssinus; Positive selection; WBC, white blood cells; Histamine release; Allergy testing; ELISA, enzyme-linked immunosorbent assay; HSA, human serum albumin; HACMG, HA with CaCl 2, MgCl 2 and glucose; Basophil; HEPES, 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid; HPLC, high-performance liquid chromatography; EDTA, ethylenediaminetetraacetic acid disodium salt; HRP, horseradish peroxidase; Magnetic beads; HA, HEPES-buffered saline with HSA; Histamine; Immunomagnetic method; Laboratory investigations; Monoclonal antibody; Allergenicity; Release; Quantitative analysis; Microassay
• DOI: 10.1016/S0022-1759(00)00169-1
• ISSN: 0022-1759

A new method for evaluating histamine release from purified basophils was developed. Basophil-containing leukocytes were directly purified from a small amount of peripheral blood using monoclonal antibody BA312-coated magnetic beads. The purified basophils still rosetted to magnetic beads maintained a normal response to anti-IgE and to dust mite allergen in comparison with the conventional method using washed leukocytes. This methodology facilitates the purification of basophils, anti-IgE- and allergen-induced histamine release, and subsequent histamine determination within only 3 h. The released histamine was analyzed by an enzyme-linked immunosorbent assay (ELISA) with a characteristic detection profile. Since all steps were performed in 96-well microplates, many clinical samples could be analyzed at the same time, permitting easy applications in routine laboratories.