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J Allergy Clin Immunol. 2004 Jun;113(6):1086-92.

Expression of alpha-tryptase and beta-tryptase by human basophils.

Author information

1
Division of Rheumatology, Allergy and Immunology, Department of Internal Medicine, Virginia Commonwealth University, PO Box 980263, Richmond, VA 23298, USA.

Abstract

BACKGROUND:

Alpha and beta-tryptase levels in serum are clinical tools for the evaluation of systemic anaphylaxis and systemic mastocytosis. Basophils and mast cells are known to produce these proteins.

OBJECTIVE:

The current study examines the effect of the alpha,beta-tryptase genotype on basophil tryptase levels and the type of tryptase stored in these cells.

METHODS:

Tryptase extracted from purified peripheral blood basophils from 20 subjects was examined by using ELISAs measuring mature and total tryptase and by using an enzymatic assay with tosyl-Gly-Pro-Lys-p-nitroanilide. Tryptase genotypes (4:0, 3:1, and 2:2 beta/alpha ratios) were assessed by using a hot-stop PCR technique with alpha,beta-tryptase-specific primers. Total alpha,beta-tryptase mRNA was measured by means of competitive RT-PCR, and ratios of alpha to beta-tryptase mRNA were measured by means of hot-stop RT-PCR.

RESULTS:

Tryptase in all but one of the basophil preparations was mature and enzymatically active. Tryptase quantities in basophils were less than 1% of those in tissue mast cells. Tryptase genotypes (beta/alpha) among the 20 donors were 4:0 in 7, 3:1 in 7, and 2:2 in 6. Tryptase protein and mRNA levels per basophil were not affected by the tryptase genotype.

CONCLUSION:

Basophils from healthy subjects contain modest amounts of mature and enzymatically active tryptase unaffected by the tryptase genotype.

PMID:
15208589
DOI:
10.1016/j.jaci.2004.02.032
[Indexed for MEDLINE]

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