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)

Christine M. Freeman, Sean Crudgington, Valerie R. Stolberg, Jeanette P. Brown, Joanne Sonstein, Neil E. Alexis, Claire M. Doerschuk, Patricia V. Basta, Elizabeth E. Carretta, David J. Couper, Annette T. Hastie, Robert J. Kaner, Wanda K. O'Neal, Robert Paine, Stephen I. Rennard, Daichi Shimbo, Prescott G. Woodruff, Michelle Zeidler, Jeffrey L. Curtis

Research output: Contribution to journalArticle

20 Citations (Scopus)

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 .

Original languageEnglish (US)
Article number19
JournalJournal of Translational Medicine
Volume13
Issue number1
DOIs
StatePublished - Jan 27 2015

Fingerprint

Immunophenotyping
Pulmonary diseases
Bronchoalveolar Lavage Fluid
Sputum
Chronic Obstructive Pulmonary Disease
Bronchoalveolar Lavage
Blood
Outcome Assessment (Health Care)
Fluids
Leukocytes
Cells
T-cells
Antibodies
Macrophages
Biomarkers
Pathology
Phenotype
Spirometry
Bronchoscopy
Data acquisition

Keywords

  • Bronchoalveolar lavage
  • COPD
  • Flow cytometry
  • Human
  • Immunophenotyping
  • Sputum

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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). / Freeman, Christine M.; Crudgington, Sean; Stolberg, Valerie R.; Brown, Jeanette P.; Sonstein, Joanne; Alexis, Neil E.; Doerschuk, Claire M.; Basta, Patricia V.; Carretta, Elizabeth E.; Couper, David J.; Hastie, Annette T.; Kaner, Robert J.; O'Neal, Wanda K.; Paine, Robert; Rennard, Stephen I.; Shimbo, Daichi; Woodruff, Prescott G.; Zeidler, Michelle; Curtis, Jeffrey L.

In: Journal of Translational Medicine, Vol. 13, No. 1, 19, 27.01.2015.

Research output: Contribution to journalArticle

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 2015, '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)', Journal of Translational Medicine, vol. 13, no. 1, 19. https://doi.org/10.1186/s12967-014-0374-z
Freeman, Christine M. ; Crudgington, Sean ; Stolberg, Valerie R. ; Brown, Jeanette P. ; Sonstein, Joanne ; Alexis, Neil E. ; Doerschuk, Claire M. ; Basta, Patricia V. ; Carretta, Elizabeth E. ; Couper, David J. ; Hastie, Annette T. ; Kaner, Robert J. ; O'Neal, Wanda K. ; Paine, Robert ; Rennard, Stephen I. ; Shimbo, Daichi ; Woodruff, Prescott G. ; Zeidler, Michelle ; Curtis, Jeffrey L. / 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). In: Journal of Translational Medicine. 2015 ; Vol. 13, No. 1.
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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 .",
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T1 - 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)

AU - Freeman, Christine M.

AU - Crudgington, Sean

AU - Stolberg, Valerie R.

AU - Brown, Jeanette P.

AU - Sonstein, Joanne

AU - Alexis, Neil E.

AU - Doerschuk, Claire M.

AU - Basta, Patricia V.

AU - Carretta, Elizabeth E.

AU - Couper, David J.

AU - Hastie, Annette T.

AU - Kaner, Robert J.

AU - O'Neal, Wanda K.

AU - Paine, Robert

AU - Rennard, Stephen I.

AU - Shimbo, Daichi

AU - Woodruff, Prescott G.

AU - Zeidler, Michelle

AU - Curtis, Jeffrey L.

PY - 2015/1/27

Y1 - 2015/1/27

N2 - 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 .

AB - 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 .

KW - Bronchoalveolar lavage

KW - COPD

KW - Flow cytometry

KW - Human

KW - Immunophenotyping

KW - Sputum

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