Evaluation of 3H‐Histamine Incorporation into Human Peripheral Blood Leukocytes
M. Barazarte, M.C. Conroy, A.L. de Weck
Abstract
M. Barazarte, M.C. Conroy, A.L. de Weck
Abstract
We have examined the use of 3H-histamine for evaluating in vitro basophil responses. Incubation of 3H-histamine for 4 h with basophil-enriched mononuclear fractions containing autologous plasma, resulted in the best incorporation of the label. Even so, less than 3% of the added label was actually recovered and other leukocytes also bound or incorporated 3H-histamine. Labeled mononuclear preparations failed to release 3H-histamine when challenged with zymosan-activated serum or anti-IgE, but endogenous histamine released under the same experimental conditions could be determined fluorometrically. The calcium ionophore, A23187, however, did cause a dose-dependent release of 3H-histamine. We conclude that 3H-histamine is not preferentially incorporated into basophil granules, and that this technique cannot be used for assessing allergen and/or IgE-mediated basophil responses.
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We have examined the use of 3H-histamine for evaluating in vitro basophil responses. Incubation of 3H-histamine for 4 h with basophil-enriched mononuclear fractions containing autologous plasma, resulted in the best incorporation of the label. Even so, less than 3% of the added label was actually recovered and other leukocytes also bound or incorporated 3H-histamine. Labeled mononuclear preparations failed to release 3H-histamine when challenged with zymosan-activated serum or anti-IgE, but endogenous histamine released under the same experimental conditions could be determined fluorometrically. The calcium ionophore, A23187, however, did cause a dose-dependent release of 3H-histamine. We conclude that 3H-histamine is not preferentially incorporated into basophil granules, and that this technique cannot be used for assessing allergen and/or IgE-mediated basophil responses.
Key concepts: Histamine, Peripheral blood, Medicine, Immunology, Peripheral, Internal medicine