Localization of eosinophil granule major basic protein in human basophils

S. J. Ackerman, G. M. Kephart, Thomas Matthew Habermann, P. R. Greipp, G. J. Gleich

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Abstract

In experiments using an immunofluorescent method to localize human eosinophil granule major basic protein (MBP) in cells and tissues, a small number of cells stained for MBP that subsequently could not be identified as eosinophils. Because the Charcot-Leyden crystal protein, another eosinophil protein, was recently identified in basophils, we tested whether MBP might also be a constituent of blood basophils. Highly purified, eosinophil-free basophil suspensions were prepared using the fluorescence-activated cell sorter (FACS) to sort basophil-containing mononuclear cell preparations stained with fluorescein-conjugated sheep IgG anti-human IgE antibody. Using these FACS-purified basophils, we found that: (a) enrichment for surface IgE-positive cells (>95% basophils) by FACS also enriched for cells staining for MBP by immunofluorescence; (b) MBP appeared to be localized in the histamine-, heparin-containing granules; (c) significant quantities of MBP were measurable by radioimmunoassay (RIA) in freeze-thaw detergent extracts of purified basophils; and (d) RIA dose-response curves for extracts of purified eosinophils and basophils had identical slopes. The MBP content of basophils from normal individuals averaged 140 ng/106 cells, whereas purified eosinophils from normal donors and patients with the hypereosinophilic syndrome averaged 4,979 and 824 ng/106 cells, respectively. MBP was also detected by immunofluorescence and RIA in cells obtained from a patient with basophil leukemia, although the MBP content for basophil leukemia cells was lower than that for normal basophils. We conclude that basophils contain a protein that is immunochemically indistinguishable from eosinophil granule MBP.

Original languageEnglish (US)
Pages (from-to)946-961
Number of pages16
JournalJournal of Experimental Medicine
Volume158
Issue number3
StatePublished - 1983

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Basophils
Eosinophils
Proteins
Radioimmunoassay
Fluorescence
human PRG2 protein
Fluorescent Antibody Technique
Leukemia
Eosinophil Major Basic Protein
Hypereosinophilic Syndrome
Fluorescein
Detergents
Immunoglobulin E
Histamine
Heparin
Sheep
Suspensions
Cell Count
Immunoglobulin G

ASJC Scopus subject areas

  • Immunology

Cite this

Ackerman, S. J., Kephart, G. M., Habermann, T. M., Greipp, P. R., & Gleich, G. J. (1983). Localization of eosinophil granule major basic protein in human basophils. Journal of Experimental Medicine, 158(3), 946-961.

Localization of eosinophil granule major basic protein in human basophils. / Ackerman, S. J.; Kephart, G. M.; Habermann, Thomas Matthew; Greipp, P. R.; Gleich, G. J.

In: Journal of Experimental Medicine, Vol. 158, No. 3, 1983, p. 946-961.

Research output: Contribution to journalArticle

Ackerman, SJ, Kephart, GM, Habermann, TM, Greipp, PR & Gleich, GJ 1983, 'Localization of eosinophil granule major basic protein in human basophils', Journal of Experimental Medicine, vol. 158, no. 3, pp. 946-961.
Ackerman, S. J. ; Kephart, G. M. ; Habermann, Thomas Matthew ; Greipp, P. R. ; Gleich, G. J. / Localization of eosinophil granule major basic protein in human basophils. In: Journal of Experimental Medicine. 1983 ; Vol. 158, No. 3. pp. 946-961.
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