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Kinetics of TH2 biomarkers in sputum of asthmatics following inhaled allergen.

Zuiker RG, Ruddy MK, Morelli N, Mogg R, Rivas VM, van Dyck K, De Lepeleire I, Tanen MR, Boot JD, Kamerling IM, Diamant Z - Eur Clin Respir J (2015)

Bottom Line: To test whether allergen-induced TH2 inflammatory markers can be reproducibly quantified by sensitive detection techniques in ultracentrifuged sputum and the effect of fluticasone (FP) on these endpoints.Allergen-induced changes in inflammatory endpoints were compared between FP and placebo using a mixed model ANCOVA.FP effectively blunted both the LAR and the inflammatory biomarkers.

View Article: PubMed Central - PubMed

Affiliation: Centre for Human Drug Research, Leiden, The Netherlands.

ABSTRACT

Background: Allergen-induced late airway response offers important pharmacodynamic targets, including T helper 2 (TH2) biomarkers. However, detection of inflammatory markers has been limited in dithiothreitol-processed sputum.

Objectives: To test whether allergen-induced TH2 inflammatory markers can be reproducibly quantified by sensitive detection techniques in ultracentrifuged sputum and the effect of fluticasone (FP) on these endpoints.

Methods: Thirteen allergic asthmatics with dual allergen-induced airway responses, documented during a single-blind placebo run-in period, participated in a double-blind, two-period crossover study. Each period consisted of three consecutive days, separated by ≥3 weeks. Following randomization, subjects inhaled FP (500 µg bid, five doses total) or placebo. On Day 2 in each study period, allergen challenge was performed and airway response measured by forced expiratory volume in 1 sec (FEV1) until 7 h post-challenge. Sputum was induced 24 h pre-allergen and 7 and 24 h post-allergen. Sputum samples were split into two portions: TH2 biomarkers were quantified by Meso Scale multiplex platform following ultracentrifugation, and cell differentials were counted on Giemsa-May-Grünwald-stained cytospins. Allergen-induced changes in inflammatory endpoints were compared between FP and placebo using a mixed model ANCOVA.

Results: Inhaled allergen induced dual airway responses in all subjects during both placebo periods with reproducible late asthmatic response (LAR) and increased sputum inflammatory biomarkers (IL-2, IL-4, IL-13, and eotaxin-1) and eosinophil counts. FP effectively blunted both the LAR and the inflammatory biomarkers.

Conclusions: Combining novel, sensitive quantification methods with ultracentrifugation allows reproducible quantification of sputum biomarkers following allergen challenge, reversed by FP. This approach allows non-invasive identification of pharmacodynamic targets for anti-asthma therapies.

No MeSH data available.


Related in: MedlinePlus

Changes in airway hyperresponsiveness 24 h pre-allergen versus 24 h post-allergen during run-in period, placebo treatment, and fluticasone treatment.
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Figure 0004: Changes in airway hyperresponsiveness 24 h pre-allergen versus 24 h post-allergen during run-in period, placebo treatment, and fluticasone treatment.

Mentions: During both placebo periods, allergen challenge increased non-specific AHR, by decreasing PC20FEV1methacholine at 24 h post-allergen by, on average, 1.18 (90% CI: 1.73, 0.64) doubling doses. In contrast, FP increased 24 h post-allergen PC20FEV1methacholine by on average 1.60 doubling doses (90% CI: 1.06, 2.15), resulting in a mean difference of 2.79 doubling doses (90% CI: 2.07, 3.51; p<0.001) between placebo and FP (Fig. 4).


Kinetics of TH2 biomarkers in sputum of asthmatics following inhaled allergen.

Zuiker RG, Ruddy MK, Morelli N, Mogg R, Rivas VM, van Dyck K, De Lepeleire I, Tanen MR, Boot JD, Kamerling IM, Diamant Z - Eur Clin Respir J (2015)

Changes in airway hyperresponsiveness 24 h pre-allergen versus 24 h post-allergen during run-in period, placebo treatment, and fluticasone treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4629766&req=5

Figure 0004: Changes in airway hyperresponsiveness 24 h pre-allergen versus 24 h post-allergen during run-in period, placebo treatment, and fluticasone treatment.
Mentions: During both placebo periods, allergen challenge increased non-specific AHR, by decreasing PC20FEV1methacholine at 24 h post-allergen by, on average, 1.18 (90% CI: 1.73, 0.64) doubling doses. In contrast, FP increased 24 h post-allergen PC20FEV1methacholine by on average 1.60 doubling doses (90% CI: 1.06, 2.15), resulting in a mean difference of 2.79 doubling doses (90% CI: 2.07, 3.51; p<0.001) between placebo and FP (Fig. 4).

Bottom Line: To test whether allergen-induced TH2 inflammatory markers can be reproducibly quantified by sensitive detection techniques in ultracentrifuged sputum and the effect of fluticasone (FP) on these endpoints.Allergen-induced changes in inflammatory endpoints were compared between FP and placebo using a mixed model ANCOVA.FP effectively blunted both the LAR and the inflammatory biomarkers.

View Article: PubMed Central - PubMed

Affiliation: Centre for Human Drug Research, Leiden, The Netherlands.

ABSTRACT

Background: Allergen-induced late airway response offers important pharmacodynamic targets, including T helper 2 (TH2) biomarkers. However, detection of inflammatory markers has been limited in dithiothreitol-processed sputum.

Objectives: To test whether allergen-induced TH2 inflammatory markers can be reproducibly quantified by sensitive detection techniques in ultracentrifuged sputum and the effect of fluticasone (FP) on these endpoints.

Methods: Thirteen allergic asthmatics with dual allergen-induced airway responses, documented during a single-blind placebo run-in period, participated in a double-blind, two-period crossover study. Each period consisted of three consecutive days, separated by ≥3 weeks. Following randomization, subjects inhaled FP (500 µg bid, five doses total) or placebo. On Day 2 in each study period, allergen challenge was performed and airway response measured by forced expiratory volume in 1 sec (FEV1) until 7 h post-challenge. Sputum was induced 24 h pre-allergen and 7 and 24 h post-allergen. Sputum samples were split into two portions: TH2 biomarkers were quantified by Meso Scale multiplex platform following ultracentrifugation, and cell differentials were counted on Giemsa-May-Grünwald-stained cytospins. Allergen-induced changes in inflammatory endpoints were compared between FP and placebo using a mixed model ANCOVA.

Results: Inhaled allergen induced dual airway responses in all subjects during both placebo periods with reproducible late asthmatic response (LAR) and increased sputum inflammatory biomarkers (IL-2, IL-4, IL-13, and eotaxin-1) and eosinophil counts. FP effectively blunted both the LAR and the inflammatory biomarkers.

Conclusions: Combining novel, sensitive quantification methods with ultracentrifugation allows reproducible quantification of sputum biomarkers following allergen challenge, reversed by FP. This approach allows non-invasive identification of pharmacodynamic targets for anti-asthma therapies.

No MeSH data available.


Related in: MedlinePlus