Increased urinary excretion of the prostaglandin D2 metabolite 9α,11β-prostaglandin F2 after aspirin challenge supports mast cell activation in aspirin-induced.

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Increased urinary excretion of the prostaglandin D2 metabolite 9α,11β-prostaglandin F2 after aspirin challenge supports mast cell activation in aspirin-induced airway obstruction Siobhán O’Sullivan, BSca,b, Barbro Dahlén, MD, PhDc, Sven-Erik Dahlén, MD, PhDa,d, Maria Kumlin, PhDb,d Journal of Allergy and Clinical Immunology Volume 98, Issue 2, Pages 421-432 (August 1996) DOI: 10.1016/S0091-6749(96)70167-7 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Accuracy of EIA. Known amounts of synthetic 9α,11β-PGF2 added to buffer (open squares) and unextracted urine (filled triangles) were measured with EIA. Accuracy was excellent, with r2 = 0.99 in both buffer and urine (mean ± SEM, n = 8). Conc, Concentration. Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Chromatographic validation of immunoreactivity. A, RP-HPLC of synthetic standard 9α,11β-PGF2 in buffer, after purification by solid-phase extraction (for details see Methods). The compound was detected as EIA immunoreactivity (filled circles) and UV absorbance (unbroken line). B, RP-HPLC of a purified urine sample collected from a patient with atopic asthma after allergen challenge. Filled circles show 9α,11β-PGF2 immunoreactivity in the fractions assessed by EIA. Losses of 9α,11β-PGF2 during purification were corrected for by use of tritiated PGF2α as internal standard. Open squares show distribution of tritiated PGF2α as determined by liquid scintillation counting. The amount of endogenous urinary 9α,11β-PGF2 was below the detection limit for UV absorbance measurements. Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Chromatographic validation of immunoreactivity. A, RP-HPLC of synthetic standard 9α,11β-PGF2 in buffer, after purification by solid-phase extraction (for details see Methods). The compound was detected as EIA immunoreactivity (filled circles) and UV absorbance (unbroken line). B, RP-HPLC of a purified urine sample collected from a patient with atopic asthma after allergen challenge. Filled circles show 9α,11β-PGF2 immunoreactivity in the fractions assessed by EIA. Losses of 9α,11β-PGF2 during purification were corrected for by use of tritiated PGF2α as internal standard. Open squares show distribution of tritiated PGF2α as determined by liquid scintillation counting. The amount of endogenous urinary 9α,11β-PGF2 was below the detection limit for UV absorbance measurements. Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 EIA analysis of 9α,11β-PGF2 (filled circles) in unpurified urine samples collected at three hourly intervals over a 24-hour period, from healthy nonasthmatic subjects (mean ± SEM, n = 5). Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 Allergen-induced urinary excretion of 9α,11β-PGF2. Mean hourly levels of 9α,11β-PGF2 (filled squares) assayed in urine samples of nine patients with atopic asthma, undergoing bronchial provocation with allergen. Arrow indicates the time of maximum drop in FEV1 (mean ± SEM, n = 9). The difference in pre- and postchallenge levels of 9α,11β-PGF2 was statistically significant (p < 0.05). Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Aspirin-induced urinary excretion of 9α,11β-PGF2. Prechallenge (open bars) and postchallenge (hatched bars) levels of 9α,11β-PGF2 in urine samples from eight patients with aspirin-intolerant asthma after challenge with lysine aspirin. A, Drug naive control session (placebo). B, After pretreatment with the LT-receptor antagonist MK-0679 (drug session). All patients received higher doses of lysine aspirin during the MK-0679 session (B). The urine sample collected immediately before inhalation challenge with lysine aspirin is defined as the prechallenge level, and the peak detected after maximal airway obstruction, as the postchallenge level (mean ± SEM). Levels of statistical significance are indicated by asterisks (Student’s t test, *p < 0.05, **p < 0.01). Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Aspirin-induced urinary excretion of 9α,11β-PGF2. Prechallenge (open bars) and postchallenge (hatched bars) levels of 9α,11β-PGF2 in urine samples from eight patients with aspirin-intolerant asthma after challenge with lysine aspirin. A, Drug naive control session (placebo). B, After pretreatment with the LT-receptor antagonist MK-0679 (drug session). All patients received higher doses of lysine aspirin during the MK-0679 session (B). The urine sample collected immediately before inhalation challenge with lysine aspirin is defined as the prechallenge level, and the peak detected after maximal airway obstruction, as the postchallenge level (mean ± SEM). Levels of statistical significance are indicated by asterisks (Student’s t test, *p < 0.05, **p < 0.01). Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 6 No increase in urinary 9α,11β-PGF2 after bronchoprovocation with histamine. Prechallenge (open bar) and postchallenge (filled bar) levels of 9α,11β-PGF2 after duplicate histamine provocations in four aspirin-intolerant patients with asthma. (The result for each individual is the mean of two different provocations, done at least 6 weeks apart). Values are expressed as means ± SEM. Journal of Allergy and Clinical Immunology 1996 98, 421-432DOI: (10.1016/S0091-6749(96)70167-7) Copyright © 1996 Mosby, Inc. Terms and Conditions