Title | Design of a multi-center immunophenotyping analysis of peripheral blood, sputum and bronchoalveolar lavage fluid in the Subpopulations and Intermediate Outcome Measures in COPD Study (SPIROMICS). |
Publication Type | Publication |
Year | 2015 |
Authors | Freeman CM, Crudgington S, Stolberg VR, Brown JP, Sonstein J, Alexis NE, Doerschuk CM, Basta PV, Carretta EE, Couper DJ, Hastie AT, Kaner RJ, O'Neal WK, Paine R, Rennard SI, Shimbo D, Woodruff PG, Zeidler M, Curtis JL |
Journal | J Transl Med |
Volume | 13 |
Pagination | 19 |
Date Published | 2015 Jan 27 |
ISSN | 1479-5876 |
Keywords | biomarkers, Bronchoalveolar Lavage Fluid, CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Cell Separation, Dendritic Cells, Flow Cytometry, Humans, Immunophenotyping, Leukocytes, Longitudinal Studies, Macrophages, phenotype, Pulmonary Disease, Chronic Obstructive, Research Design, Sample Size, smoking, Spirometry, Sputum |
Abstract | BACKGROUND: Subpopulations and Intermediate Outcomes in COPD Study (SPIROMICS) is a multi-center longitudinal, observational study to identify novel phenotypes and biomarkers of chronic obstructive pulmonary disease (COPD). In a subset of 300 subjects enrolled at six clinical centers, we are performing flow cytometric analyses of leukocytes from induced sputum, bronchoalveolar lavage (BAL) and peripheral blood. To minimize several sources of variability, we use a "just-in-time" design that permits immediate staining without pre-fixation of samples, followed by centralized analysis on a single instrument.METHODS: The Immunophenotyping Core prepares 12-color antibody panels, which are shipped to the six Clinical Centers shortly before study visits. Sputum induction occurs at least two weeks before a bronchoscopy visit, at which time peripheral blood and bronchoalveolar lavage are collected. Immunostaining is performed at each clinical site on the day that the samples are collected. Samples are fixed and express shipped to the Immunophenotyping Core for data acquisition on a single modified LSR II flow cytometer. Results are analyzed using FACS Diva and FloJo software and cross-checked by Core scientists who are blinded to subject data.RESULTS: Thus far, a total of 152 sputum samples and 117 samples of blood and BAL have been returned to the Immunophenotyping Core. Initial quality checks indicate useable data from 126 sputum samples (83%), 106 blood samples (91%) and 91 BAL samples (78%). In all three sample types, we are able to identify and characterize the activation state or subset of multiple leukocyte cell populations (including CD4+ and CD8+ T cells, B cells, monocytes, macrophages, neutrophils and eosinophils), thereby demonstrating the validity of the antibody panel.CONCLUSIONS: Our study design, which relies on bi-directional communication between clinical centers and the Core according to a pre-specified protocol, appears to reduce several sources of variability often seen in flow cytometric studies involving multiple clinical sites. Because leukocytes contribute to lung pathology in COPD, these analyses will help achieve SPIROMICS aims of identifying subgroups of patients with specific COPD phenotypes. Future analyses will correlate cell-surface markers on a given cell type with smoking history, spirometry, airway measurements, and other parameters.TRIAL REGISTRATION: This study was registered with ClinicalTrials.gov as NCT01969344 . |
DOI | 10.1186/s12967-014-0374-z |
Alternate Journal | J Transl Med |
PubMed ID | 25622723 |
PubMed Central ID | PMC4314767 |
Grant List | HHSN268200900017C / / PHS HHS / United States T32 HL007749 / HL / NHLBI NIH HHS / United States HHSN268200900019C / HL / NHLBI NIH HHS / United States HHSN268200900009C / WH / WHI NIH HHS / United States HHSN268200900015C / / PHS HHS / United States HHSN268200900020C / / PHS HHS / United States HHSN268200900018C / / PHS HHS / United States HHSN268200900015C / HL / NHLBI NIH HHS / United States HHSN268200900016C / HL / NHLBI NIH HHS / United States HHSN268200900014C / / PHS HHS / United States NHLBI-HR-08-08 / HR / NHLBI NIH HHS / United States HHSN268200900018C / HL / NHLBI NIH HHS / United States HHSN268200900017C / HL / NHLBI NIH HHS / United States HHSN268200900020C / HL / NHLBI NIH HHS / United States HHSN268200900013C / HL / NHLBI NIH HHS / United States HHSN2682009000019C / / PHS HHS / United States HHSN268200900013C / / PHS HHS / United States HHSN268200900014C / HL / NHLBI NIH HHS / United States HHSN268200900016C / / PHS HHS / United States |
Design of a multi-center immunophenotyping analysis of peripheral blood, sputum and bronchoalveolar lavage fluid in the Subpopulations and Intermediate Outcome Measures in COPD Study (SPIROMICS).
MS#:
MS017
Manuscript Full Title:
Design of a multi-center immunophenotyping analysis of peripheral blood, sputum and bronchoalveolar lavage fluid in the Subpopulations and Intermediate Outcome Measures in COPD Study (SPIROMICS).
ECI:
Manuscript Status:
Published and Public