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Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects

PURPOSE: The purpose of this study was to investigate the reproducibility of fluid-phase measurements in PBS-treated sputum supernatant, processed using the two-step method, of healthy and stable COPD individuals. METHODS: Nine healthy subjects and 23 stable COPD patients provided sputum twice withi...

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Autores principales: Wang, Fengyan, Liang, Zhenyu, Yang, Yuqiong, Zhou, Luqian, Guan, Lili, Wu, Weiliang, Jiang, Mei, Shi, Weijuan, Deng, Kuimiao, Chen, Jianhua, Chen, Rongchang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469484/
https://www.ncbi.nlm.nih.gov/pubmed/31043775
http://dx.doi.org/10.2147/COPD.S187661
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author Wang, Fengyan
Liang, Zhenyu
Yang, Yuqiong
Zhou, Luqian
Guan, Lili
Wu, Weiliang
Jiang, Mei
Shi, Weijuan
Deng, Kuimiao
Chen, Jianhua
Chen, Rongchang
author_facet Wang, Fengyan
Liang, Zhenyu
Yang, Yuqiong
Zhou, Luqian
Guan, Lili
Wu, Weiliang
Jiang, Mei
Shi, Weijuan
Deng, Kuimiao
Chen, Jianhua
Chen, Rongchang
author_sort Wang, Fengyan
collection PubMed
description PURPOSE: The purpose of this study was to investigate the reproducibility of fluid-phase measurements in PBS-treated sputum supernatant, processed using the two-step method, of healthy and stable COPD individuals. METHODS: Nine healthy subjects and 23 stable COPD patients provided sputum twice within 6 days. A two-step sputum processing method was used to obtain PBS-treated supernatant and sputum cells. Soluble protein markers and IgG and IgM autoantibody profiles in PBS supernatant were analyzed using customized microarrays. Repeatability of measurements was assessed by paired-sample testing and an intraclass correlation coefficient, then graphically reported by Bland–Altman plot. RESULTS: There was no significant difference between the repeated detection of 8/10 types of soluble protein markers, all 13 types of IgG autoantibodies, and 12/13 types of corresponding IgM autoantibodies in PBS supernatant. The repeatability of measurements in PBS supernatant was substantial to very good for interleukin 6 (IL6), IL8, IL13, IL10, IL33, vascular endothelial growth factor, soluble receptor for advanced glycation end-products, and tumor necrosis factor-α; for IgG autoantibodies against aggrecan, centromere protein B (CENP-B), collagen II, collagen IV, cytochrome C, elastin, heat shock protein 47 (HSP47), HSP70, and La/Sjögren syndrome type B antigen; for IgM autoantibodies against CENP-B, collagen I, collagen II, collagen IV, cytokeratin 18, and HSP70; and for sputum neutrophils, macrophages and eosinophils count. Bland–Altman plots suggested good consistency within repeated measurements. Stable COPD patients differed from healthy subjects in the proportion of neutrophils and eosinophils; relative fluorescence intensity of anti-cytochrome C IgG, anti-aggrecan IgM, and anti-cytochrome C IgM. There was a significant positive correlation for stable COPD patients between sputum anti-collagen II IgG and post-bronchodilator FEV(1)%. CONCLUSION: We confirmed fluid-phase measurements in PBS-treated sputum supernatant by high-throughput techniques with good repeatability. We demonstrated the presence of IgG and IgM autoantibodies to multiple antigens in the airways of COPD patients.
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spelling pubmed-64694842019-05-01 Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects Wang, Fengyan Liang, Zhenyu Yang, Yuqiong Zhou, Luqian Guan, Lili Wu, Weiliang Jiang, Mei Shi, Weijuan Deng, Kuimiao Chen, Jianhua Chen, Rongchang Int J Chron Obstruct Pulmon Dis Original Research PURPOSE: The purpose of this study was to investigate the reproducibility of fluid-phase measurements in PBS-treated sputum supernatant, processed using the two-step method, of healthy and stable COPD individuals. METHODS: Nine healthy subjects and 23 stable COPD patients provided sputum twice within 6 days. A two-step sputum processing method was used to obtain PBS-treated supernatant and sputum cells. Soluble protein markers and IgG and IgM autoantibody profiles in PBS supernatant were analyzed using customized microarrays. Repeatability of measurements was assessed by paired-sample testing and an intraclass correlation coefficient, then graphically reported by Bland–Altman plot. RESULTS: There was no significant difference between the repeated detection of 8/10 types of soluble protein markers, all 13 types of IgG autoantibodies, and 12/13 types of corresponding IgM autoantibodies in PBS supernatant. The repeatability of measurements in PBS supernatant was substantial to very good for interleukin 6 (IL6), IL8, IL13, IL10, IL33, vascular endothelial growth factor, soluble receptor for advanced glycation end-products, and tumor necrosis factor-α; for IgG autoantibodies against aggrecan, centromere protein B (CENP-B), collagen II, collagen IV, cytochrome C, elastin, heat shock protein 47 (HSP47), HSP70, and La/Sjögren syndrome type B antigen; for IgM autoantibodies against CENP-B, collagen I, collagen II, collagen IV, cytokeratin 18, and HSP70; and for sputum neutrophils, macrophages and eosinophils count. Bland–Altman plots suggested good consistency within repeated measurements. Stable COPD patients differed from healthy subjects in the proportion of neutrophils and eosinophils; relative fluorescence intensity of anti-cytochrome C IgG, anti-aggrecan IgM, and anti-cytochrome C IgM. There was a significant positive correlation for stable COPD patients between sputum anti-collagen II IgG and post-bronchodilator FEV(1)%. CONCLUSION: We confirmed fluid-phase measurements in PBS-treated sputum supernatant by high-throughput techniques with good repeatability. We demonstrated the presence of IgG and IgM autoantibodies to multiple antigens in the airways of COPD patients. Dove Medical Press 2019-04-11 /pmc/articles/PMC6469484/ /pubmed/31043775 http://dx.doi.org/10.2147/COPD.S187661 Text en © 2019 Wang et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.
spellingShingle Original Research
Wang, Fengyan
Liang, Zhenyu
Yang, Yuqiong
Zhou, Luqian
Guan, Lili
Wu, Weiliang
Jiang, Mei
Shi, Weijuan
Deng, Kuimiao
Chen, Jianhua
Chen, Rongchang
Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title_full Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title_fullStr Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title_full_unstemmed Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title_short Reproducibility of fluid-phase measurements in PBS-treated sputum supernatant of healthy and stable COPD subjects
title_sort reproducibility of fluid-phase measurements in pbs-treated sputum supernatant of healthy and stable copd subjects
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469484/
https://www.ncbi.nlm.nih.gov/pubmed/31043775
http://dx.doi.org/10.2147/COPD.S187661
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