A new radioimmunoassay for human mast cell tryptase using monoclonal antibodies

Enander, I.; Matsson, P.; Nystrand, J; Andersson, Ann‐Sofie; Eklund, Elisabeth; Bradford, Timothy R.; Schwartz, Lawrence B.

doi:10.1016/0022-1759(91)90062-k

Cited by 102 publications

(39 citation statements)

References 24 publications

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“…The mouse mAbs against human tryptase called G5, B 12, G3, and G4, each recognizing a distinct epitope, were prepared and purified as described (8,9,13,14). Antibodies to be biotinylated (G5, G3, and G4) were dialyzed in PBS, diluted to a concentration of 2 mg/ml with 0.1 M NaHCO3, incubated with a fivefold molar excess of NHS-LC-biotin for -4 h at room temperature, treated with 1/2 vol of 0.1 M Tris, pH 8.0, 0.15 M NaCl, concentrated by ultrafiltration, dialyzed against PBS containing 20% glycerol and 0.05% NaN3, and stored frozen at -70'C.…”

Section: Methodsmentioning

confidence: 99%

“…Antibodies to be biotinylated (G5, G3, and G4) were dialyzed in PBS, diluted to a concentration of 2 mg/ml with 0.1 M NaHCO3, incubated with a fivefold molar excess of NHS-LC-biotin for -4 h at room temperature, treated with 1/2 vol of 0.1 M Tris, pH 8.0, 0.15 M NaCl, concentrated by ultrafiltration, dialyzed against PBS containing 20% glycerol and 0.05% NaN3, and stored frozen at -70'C. The immunoassay using G5 for capture and 1251I-G4 for detection (8), and the one using B 12 for capture and biotinylated G4 and G3 for detection (9) were each performed as described. In each case, tryptase purified to homogeneity from human lung and measured by the bicinchoninic acid-modified Lowry assay for protein (15) was used as the tryptase standard.…”

Section: Methodsmentioning

confidence: 99%

“…Basophils are the only other cell type with detectable tryptase, but levels are several hundredfold lower than in mast cells, and represent a negligible amount (7). Two immunoassays for measuring tryptase are currently in use in our laboratory and will be referred to as the G5 (8) and B12 (9) immunoassays. The G5 immunoassay used the G5 mAb for capture and the noncompeting G4 mAb for detection.…”

Section: Introductionmentioning

confidence: 99%

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The alpha form of human tryptase is the predominant type present in blood at baseline in normal subjects and is elevated in those with systemic mastocytosis.

Schwartz¹,

Sakai²,

Bradford³

et al. 1995

J. Clin. Invest.

342

251

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Tryptase, a protease produced by all mast cells, was evaluated as a clinical marker of systemic mastocytosis. Two sandwich immunoassays were evaluated, one which used the mAb G5 for capture, the other which used B12 for capture. The B12 capture assay measured both recombinant a-and .-tryptase, whereas the G5 capture assay measured primarily recombinant f8-tryptase. G5 binds with low affinity to both recombinant c-tryptase and tryptase in blood from normal and nonacute mastocytosis subjects, and binds with high affinity to recombinant ,B-tryptase, tryptase in serum during anaphylaxis, and tryptase stored in mast cell secretory granules. B12 recognizes all of these forms of tryptase with high affinity. As reported previously, during systemic anaphylaxis in patients without known mastocytosis, the ratio of B12-to G5-measured tryptase was always < 5 and approached unity (Schwartz L. B., T. R. Bradford, C. Rouse, A.-M. Irani, G. Rasp, J. K. Van der Zwan and P.-W. G. . In this report, most mastocytosis patients with systemic disease have B12-measured tryptase levels that are elevated (> 20 ng/ml) and are at least 10-fold greater than the corresponding G5-measured tryptase level. Most of those subjects with B12-measured tryptase levels of < 20 ng/ml had only cutaneous manifestations. The B12 assay for a-tryptase and ,3-tryptase, particularly when performed in conjunction with the G5 assay for j8-tryptase, provides a more precise measure of mast cell involvement than currently available assessments, a promising potential screening test for systemic mastocytosis and may provide an improved means to follow disease progression and response to therapy. (J. Clin. Invest. 1995Invest. . 96:2702Invest. -2710

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The alpha form of human tryptase is the predominant type present in blood at baseline in normal subjects and is elevated in those with systemic mastocytosis.

Schwartz¹,

Sakai²,

Bradford³

et al. 1995

J. Clin. Invest.

342

251

View full text Add to dashboard Cite

show abstract

“…In selected cases with high levels of total tryptase, ␤-tryptase levels were measured by enzyme-linked immunosorbent assay (ELISA). 39 Histamine was measured in whole blood lysates (generated by freeze thawing) by a commercial radioimmunoassay (Immunotech, Marseille, France).…”

Section: Measurements Of Tryptase and Histaminementioning

confidence: 99%

Expression of mast cell tryptase by myeloblasts in a group of patients with acute myeloid leukemia

Sperr

Jordan

Baghestanian

et al. 2001

Blood

Self Cite

133

105

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“…Kits for analysis of ECP [23], MPO [24], hyaluronan [25] and tryptase [26] (Pharmacia Diagnostics AB, Uppsala, Sweden) were used according to the instructions of the manufacturers.…”

Section: Analysis Of Inflammatory Markersmentioning

confidence: 99%

Bronchial brush biopsies for studies of epithelial inflammation in stable asthma and nonobstructive chronic bronchitis

Riise

Åndersson²,

Ahlstedt³

et al. 1996

Eur Respir J

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Recently, bronchial brush biopsy (BBB) has been introduced as a complimentary method to bronchial forceps biopsy for the study of bronchial epithelial cells. We wanted to determine whether epithelial inflammatory cells in bronchial brush biopsies can reflect mucosal inflammation assessed indirectly by levels of cellular activation markers in bronchial lavage fluid.We studied 15 healthy controls, 11 asthmatics with regular steroid inhalation therapy, 13 asthmatics without steroids, and 10 smokers with nonobstructive chronic bronchitis. Differential counts of epithelial and inflammatory cells were made from the BBB material. Bronchial lavage levels of eosinophil cationic protein (ECP), myeloperoxidase (MPO), tryptase, hyaluronan and interleukin-8 (IL-8) were measured as indirect markers for inflammatory cell activation.We found an increased percentage of eosinophil granulocytes in the BBB from the steroid-untreated asthmatic patients (1.16%) in comparison to the other groups (0.11%, 0.09% and 0.02%, respectively; p<0.01). In the steroid-untreated asthmatic patients, the percentage of eosinophils correlated with ECP in bronchial lavage fluid (r=0.73; p<0.01), indicating that the BBB method can reflect the degree of eosinophilic activation. A negative correlation was found for the percentage of eosinophils in BBB with levels of provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second (PC20) for the asthmatic patients in the study (r= -0.67; p<0.003).The bronchial brush biopsy method appears to give information on the changes present in superficial bronchial epithelium in inflammatory airways disease. These changes appear to relate to the degree of inflammatory activity and disease severity in asthma.

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A new radioimmunoassay for human mast cell tryptase using monoclonal antibodies

Cited by 102 publications

References 24 publications

The alpha form of human tryptase is the predominant type present in blood at baseline in normal subjects and is elevated in those with systemic mastocytosis.

The alpha form of human tryptase is the predominant type present in blood at baseline in normal subjects and is elevated in those with systemic mastocytosis.

Expression of mast cell tryptase by myeloblasts in a group of patients with acute myeloid leukemia

Bronchial brush biopsies for studies of epithelial inflammation in stable asthma and nonobstructive chronic bronchitis

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