Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes
Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins i...
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| Published in: | Journal of allergy and clinical immunology Vol. 125; no. 5; pp. 1028 - 1036.e13 |
|---|---|
| Main Authors: | , , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
New York, NY
Mosby, Inc
01.05.2010
Elsevier Elsevier Limited |
| Subjects: | |
| ISSN: | 0091-6749, 1097-6825, 1097-6825 |
| Online Access: | Get full text |
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| Abstract | Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.
We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.
This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.
Stratified by sputum granulocytes, <2% or ≥2% eosinophils and <40% or ≥40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily β-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with ≥40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1β, and macrophage inflammatory protein 3α/CCL20 for those with ≥40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.
Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. |
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| AbstractList | Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. Objective - We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants. Methods - This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations. Results - Stratified by sputum granulocytes, <2% or >=2% eosinophils and <40% or >=40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta -agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with >=40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1 beta , and macrophage inflammatory protein 3 alpha /CCL20 for those with >=40% neutrophils; these mediators demonstrated positive associations with neutrophil counts. Conclusion - Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.BACKGROUNDPatients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.OBJECTIVEWe hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants.This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.METHODSThis hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations.Stratified by sputum granulocytes, <2% or > or = 2% eosinophils and <40% or > or = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with > or = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those with > or = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.RESULTSStratified by sputum granulocytes, <2% or > or = 2% eosinophils and <40% or > or = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with > or = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those with > or = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts.Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils.CONCLUSIONCombined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants. This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations. Stratified by sputum granulocytes, <2% or ≥2% eosinophils and <40% or ≥40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily β-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with ≥40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1β, and macrophage inflammatory protein 3α/CCL20 for those with ≥40% neutrophils; these mediators demonstrated positive associations with neutrophil counts. Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. Background Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. Objective We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants. Methods This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations. Results Stratified by sputum granulocytes, <2% or >=2% eosinophils and <40% or >=40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily β-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with >=40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1β, and macrophage inflammatory protein 3α/CCL20 for those with >=40% neutrophils; these mediators demonstrated positive associations with neutrophil counts. Conclusion Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. Background Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. Objective We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants. Methods This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations. Results Stratified by sputum granulocytes, <2% or ≥2% eosinophils and <40% or ≥40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily β-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with ≥40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1β, and macrophage inflammatory protein 3α/CCL20 for those with ≥40% neutrophils; these mediators demonstrated positive associations with neutrophil counts. Conclusion Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. We hypothesized that inflammatory granulocytes in sputum may identify specific asthma severity phenotypes and are associated with different patterns of inflammatory proteins in sputum supernatants. This hypothesis was tested in 242 patients with asthma enrolled in the Severe Asthma Research Program who provided sputum samples for cell count, differential cell determinations, cell lysates for Western blot, and supernatant analyses by inflammatory protein microarrays and ELISAs. ANOVA and multiple linear regression models tested mediator associations. Stratified by sputum granulocytes, <2% or > or = 2% eosinophils and <40% or > or = 40% neutrophils, subjects with both increased eosinophils and neutrophils had the lowest lung function and increased symptoms and health care use. Subjects with elevated eosinophils with or without increased neutrophils had significantly increased fraction exhaled nitric oxide (FeNO) and serum eosinophils and greater frequency of daily beta-agonist use. Microarray data stratified by granulocytes revealed 25 to 28 inflammatory proteins increased >2-fold in sputa with > or = 40% neutrophils. Microarray analyses stratified by severity of asthma identified 6 to 9 proteins increased >2-fold in sputa in subjects with severe asthma compared with nonsevere asthma. ELISA data stratified by sputum granulocytes showed significant increases in brain-derived neurotrophic factor, IL-1beta, and macrophage inflammatory protein 3alpha/CCL20 for those with > or = 40% neutrophils; these mediators demonstrated positive associations with neutrophil counts. Combined increased sputum eosinophils and neutrophils identified patients with asthma with the lowest lung function, worse asthma control, and increased symptoms and health care requirements. Inflammatory protein analyses of sputum supernatants found novel mediators increased in patients with asthma, predominantly associated with increased sputum neutrophils. |
| Author | Bleecker, Eugene R. Li, Huashi Hastie, Annette T. Meyers, Deborah A. Vestal, Penny L. Peters, Stephen P. Moore, Wendy C. |
| Author_xml | – sequence: 1 givenname: Annette T. surname: Hastie fullname: Hastie, Annette T. email: ahastie@wfubmc.edu – sequence: 2 givenname: Wendy C. surname: Moore fullname: Moore, Wendy C. – sequence: 3 givenname: Deborah A. surname: Meyers fullname: Meyers, Deborah A. – sequence: 4 givenname: Penny L. surname: Vestal fullname: Vestal, Penny L. – sequence: 5 givenname: Huashi surname: Li fullname: Li, Huashi – sequence: 6 givenname: Stephen P. surname: Peters fullname: Peters, Stephen P. – sequence: 7 givenname: Eugene R. surname: Bleecker fullname: Bleecker, Eugene R. |
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| Keywords | Asthma phenotypes TNFSF14 BDNF CCL20 EGF PARC SARP CXCL13 FVC BLC ICS MIP-3α MCP protein microarrays CCL18 SAM Forced vital capacity Epidermal growth factor B-lymphocyte chemoattractant/CXCL13 Severe Asthma Research Program of the National Heart, Lung, and Blood Institute Inhaled corticosteroid Macrophage inflammatory protein 3α/CCL20 TNF superfamily factor 14/LIGHT, Lymphotoxin homologous, Inducible expression, competes with HSV Glycoprotein D for HVEM receptor on T lymphocytes Pulmonary and regulated chemokine/CCL18 Brain-derived neurotrophic factor Monocyte chemoattractant protein Significance Analysis of Microarrays Human Lung disease Immunopathology Chemokine CC Exodus Respiratory disease Granulocyte LIGHT ligand CXCLl3 Inflammation Protein microarray CC chemokine Protein Asthma Phenotype Immunology Sputum Bronchus disease Obstructive pulmonary disease |
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| Snippet | Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.
We hypothesized that inflammatory... Background Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. Objective We... Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma.BACKGROUNDPatients with severe... Patients with severe asthma have increased granulocytes in their sputum compared with patients with mild to moderate asthma. Objective - We hypothesized that... |
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| SubjectTerms | Adolescent Adult Allergy and Immunology Asthma Asthma - physiopathology Asthma phenotypes BDNF Biological and medical sciences CCL18 CCL20 Child Child, Preschool Chronic obstructive pulmonary disease, asthma CXCL13 Enzyme-Linked Immunosorbent Assay Female Fundamental and applied biological sciences. Psychology Fundamental immunology Granulocytes - cytology Humans Immunopathology Inflammation - immunology Inflammation - physiopathology Inflammation Mediators - analysis Linear Models Male Medical sciences Meetings Methods Phenotype Pneumology Protein Array Analysis protein microarrays Proteins Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis Severity of Illness Index Sputum - cytology TNFSF14 Young Adult |
| Title | Analyses of asthma severity phenotypes and inflammatory proteins in subjects stratified by sputum granulocytes |
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