Expression of ⍺-tryptase and β-tryptase by human basophils - 25/08/11
, Hae-Ki Min, PhD , Yoshihiro Fukuoka, PhD, Han-Zhang Xia, MS, Lawrence B. Schwartz, MD, PhD ⁎Reprint requests: Lawrence B. Schwartz, MD, PhD, Division of Rheumatology, Allergy and Immunology, Department of Internal Medicine, Virginia Commonwealth University, PO Box 980263, Richmond, VA 23298.Richmond, Va
Résumé |
Background |
⍺ and β-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 ⍺,β-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 β/⍺ ratios) were assessed by using a hot-stop PCR technique with ⍺,β-tryptase–specific primers. Total ⍺,β-tryptase mRNA was measured by means of competitive RT-PCR, and ratios of ⍺ to β-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 (β/⍺) 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.
Le texte complet de cet article est disponible en PDF.Key words : Basophil, tryptase
Abbreviations used : G3PDH, Mac-6, MES
Plan
| Supported by grants from the National Institutes of Health (AI20487 and AI27517). |
Vol 113 - N° 6
P. 1086-1092 - juin 2004 Retour au numéro
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