Analyses of Asthma Severity Phenotypes and Inflammatory Proteins in Subjects Stratified by Sputum Granulocytes

Overview

Journal J Allergy Clin Immunol

Specialty Allergy & Immunology

Date 2010 Apr 20

PMID 20398920

Citations 204

Authors

Annette T Hastie

Wendy C Moore

Deborah A Meyers

Penny L Vestal

Huashi Li

Stephen P Peters

Eugene R Bleecker

Affiliations

Soon will be listed here.

Abstract

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 > 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.

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.

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References

Moore W, Bleecker E, Curran-Everett D, Erzurum S, Ameredes B, Bacharier L . Characterization of the severe asthma phenotype by the National Heart, Lung, and Blood Institute's Severe Asthma Research Program. J Allergy Clin Immunol. 2007; 119(2):405-13. PMC: 2837934. DOI: 10.1016/j.jaci.2006.11.639. View

Haldar P, Pavord I, Shaw D, Berry M, Thomas M, Brightling C . Cluster analysis and clinical asthma phenotypes. Am J Respir Crit Care Med. 2008; 178(3):218-224. PMC: 3992366. DOI: 10.1164/rccm.200711-1754OC. View

Simpson J, Timmins N, Fakes K, Talbot P, Gibson P . Effect of saliva contamination on induced sputum cell counts, IL-8 and eosinophil cationic protein levels. Eur Respir J. 2004; 23(5):759-62. DOI: 10.1183/09031936.04.00043104a. View

Simpson J, Scott R, Boyle M, Gibson P . Differential proteolytic enzyme activity in eosinophilic and neutrophilic asthma. Am J Respir Crit Care Med. 2005; 172(5):559-65. DOI: 10.1164/rccm.200503-369OC. View

Polosa R, Puddicombe S, Krishna M, Tuck A, Howarth P, Holgate S . Expression of c-erbB receptors and ligands in the bronchial epithelium of asthmatic subjects. J Allergy Clin Immunol. 2002; 109(1):75-81. DOI: 10.1067/mai.2002.120274. View

Lemiere C, Ernst P, Olivenstein R, Yamauchi Y, Govindaraju K, Ludwig M . Airway inflammation assessed by invasive and noninvasive means in severe asthma: eosinophilic and noneosinophilic phenotypes. J Allergy Clin Immunol. 2006; 118(5):1033-9. DOI: 10.1016/j.jaci.2006.08.003. View

Kim H, Kim C, Iijima K, Kobayashi T, Kita H . Protein microarray analysis in patients with asthma: elevation of the chemokine PARC/CCL18 in sputum. Chest. 2008; 135(2):295-302. PMC: 2835338. DOI: 10.1378/chest.08-0962. View

. Proceedings of the ATS workshop on refractory asthma: current understanding, recommendations, and unanswered questions. American Thoracic Society. Am J Respir Crit Care Med. 2000; 162(6):2341-51. DOI: 10.1164/ajrccm.162.6.ats9-00. View

Fox A, Patel H, Barnes P, Belvisi M . Release of nerve growth factor by human pulmonary epithelial cells: role in airway inflammatory diseases. Eur J Pharmacol. 2001; 424(2):159-62. DOI: 10.1016/s0014-2999(01)01138-4. View

10.

Fahy J, Boushey H . Effect of low-dose beclomethasone dipropionate on asthma control and airway inflammation. Eur Respir J. 1998; 11(6):1240-7. DOI: 10.1183/09031936.98.11061240. View

11.

Fulkerson P, Zimmermann N, Hassman L, Finkelman F, Rothenberg M . Pulmonary chemokine expression is coordinately regulated by STAT1, STAT6, and IFN-gamma. J Immunol. 2004; 173(12):7565-74. DOI: 10.4049/jimmunol.173.12.7565. View

12.

Virchow J, Julius P, Lommatzsch M, Luttmann W, Renz H, Braun A . Neurotrophins are increased in bronchoalveolar lavage fluid after segmental allergen provocation. Am J Respir Crit Care Med. 1998; 158(6):2002-5. DOI: 10.1164/ajrccm.158.6.9803023. View

13.

Thomas R, Green R, Brightling C, Birring S, Parker D, Wardlaw A . The influence of age on induced sputum differential cell counts in normal subjects. Chest. 2004; 126(6):1811-4. DOI: 10.1378/chest.126.6.1811. View

14.

Arend W, Palmer G, Gabay C . IL-1, IL-18, and IL-33 families of cytokines. Immunol Rev. 2008; 223:20-38. DOI: 10.1111/j.1600-065X.2008.00624.x. View

15.

Moore W, Meyers D, Wenzel S, Teague W, Li H, Li X . Identification of asthma phenotypes using cluster analysis in the Severe Asthma Research Program. Am J Respir Crit Care Med. 2009; 181(4):315-23. PMC: 2822971. DOI: 10.1164/rccm.200906-0896OC. View

16.

Reibman J, Hsu Y, Chen L, Bleck B, Gordon T . Airway epithelial cells release MIP-3alpha/CCL20 in response to cytokines and ambient particulate matter. Am J Respir Cell Mol Biol. 2003; 28(6):648-54. DOI: 10.1165/rcmb.2002-0095OC. View

17.

Fahy J, Boushey H, Lazarus S, Mauger E, CHERNIACK R, Chinchilli V . Safety and reproducibility of sputum induction in asthmatic subjects in a multicenter study. Am J Respir Crit Care Med. 2001; 163(6):1470-5. DOI: 10.1164/ajrccm.163.6.9901105. View

18.

Pichavant M, Charbonnier A, Taront S, Brichet A, Wallaert B, Pestel J . Asthmatic bronchial epithelium activated by the proteolytic allergen Der p 1 increases selective dendritic cell recruitment. J Allergy Clin Immunol. 2005; 115(4):771-8. DOI: 10.1016/j.jaci.2004.11.043. View

19.

Simpson J, Grissell T, Douwes J, Scott R, Boyle M, Gibson P . Innate immune activation in neutrophilic asthma and bronchiectasis. Thorax. 2006; 62(3):211-8. PMC: 2117164. DOI: 10.1136/thx.2006.061358. View

20.

Green R, Brightling C, Woltmann G, Parker D, Wardlaw A, Pavord I . Analysis of induced sputum in adults with asthma: identification of subgroup with isolated sputum neutrophilia and poor response to inhaled corticosteroids. Thorax. 2002; 57(10):875-9. PMC: 1746199. DOI: 10.1136/thorax.57.10.875. View