ArticleCytometry. Part B, Clinical cytometry2008
Standardized method to minimize variability in a functional P2X(7) flow cytometric assay for a multi-center clinical trial.
Article in Cytometry. Part B, Clinical cytometry, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
7 citing papers in PubMed, 13 citations in OpenAlex.
- Oxidized-ATP Attenuates Kidney Allograft Rejection By Inhibiting T-Cell, B-Cell, and Macrophage Activity.Kidney360 · 2020Article
- Altered donor P2X7 activity in human leukocytes correlates with P2RX7 genotype but does not affect the development of graft-versus-host disease in humanised mice.Purinergic signalling · 2019Article
- P2X7-regulated protection from exacerbations and loss of control is independent of asthma maintenance therapy.American journal of respiratory and critical care medicine · 2013Article
- Protection from asthma in a high-risk birth cohort by attenuated P2X(7) function.The Journal of allergy and clinical immunology · 2012Article
- Characterizing the presence and sensitivity of the P2X7 receptor in different compartments of the gut.Journal of innate immunity · 2012Review
- Leukocytes are primed in peripheral blood for activation during term and preterm labour.Molecular human reproduction · 2009Article
- Attenuated P2X7 pore function as a risk factor for virus-induced loss of asthma control.American journal of respiratory and critical care medicine · 2009Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundFlow cytometric analysis of human P2X(7) pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial.
methodsCD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X(7) agonist (BzATP) or control, followed by the addition of PI after closure of the P2X(7) pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating reliable processing of samples with varied ages.
resultsThe median YO-PRO-1 fluorescence of BzATP-treated samples had less variability when collected by the bead-adjusted method and was less influenced by the compensation strategy used. The average day-to-day coefficient of variance for assessments of P2X(7) pore activity by this method was 0.11 +/- 0.04, and the exclusion of nonviable cells was found to accommodate samples aged up to 4 days after phlebotomy. The bead-adjusted set-up method produced measurements differing by only 2.0% +/- 1.5% on two analog cytometers, and within similar decades when comparing analog to digital instruments.
conclusionsThese results provide a standardized method for quantitative flow cytometric analysis of P2X(7) receptor phenotypes in blood monocytes with minimal intralaboratory variation and potential for interlaboratory comparisons that can greatly facilitate multicenter functional genomic clinical studies.
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