ABSTRACTS Author disclosures are available with the text of this abstract at www.atsjournals.org.
could contribute to disease prevention and could open up new treatment strategies.
(Received in original form July 18, 2013; accepted in final form July 26, 2013)
Author disclosures are available with the text of this abstract at www.atsjournals.org.
Correspondence and requests for reprints should be addressed to Candace Cynclare Clay, University of California Davis, California National Primate Research Center, Respiratory Diseases Unit, County Road 98, Primate Center, Davis, CA 95616. E-mail:
[email protected] Ann Am Thorac Soc Vol 11, Supplement 1, pp S75–S76, Jan 2014 Copyright © 2014 by the American Thoracic Society Internet address: www.atsjournals.org
Microbes and Microbial Products in Cigarette Smoke
(Received in original form August 15, 2013; accepted in final form September 3, 2013) Correspondence and requests for reprints should be addressed to Gillina Bezemer, M.Sc., Utrecht University, Pharmacology, David de Wied Building, 2nd floor, Universiteitslaan 99, Utrecht, 3584CG The Netherlands. E-mail:
[email protected] Ann Am Thorac Soc Vol 11, Supplement 1, p S76, Jan 2014 Copyright © 2014 by the American Thoracic Society Internet address: www.atsjournals.org
Implications for Chronic Obstructive Pulmonary Disease Gillina Bezemer1, Eric Jubinville2, Esmail Mortaz3, Aletta Kraneveld3, Johan Garssen3, Caroline Duchaine2, and Gert Folkerts3
Novosphingobium spp. in Chronic Obstructive Pulmonary Disease in Humans and Subacute Lung Inflammation in Mice
1 Department of Pharmacology, and 3Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands; and 2Centre de Recherche de l’Institut Universitaire de Cardiologie et de Pneumologie de Quebec, Universite Laval, Quebec City, Quebec, Canada
Alleluiah Rutebemberwa1, Mark Stevens2, Mario Perez1, Lynelle Smith1, Linda Sanders1, Rubin Tuder1, and J. Kirk Harris2
There is growing evidence that changes in the lung microbiome are involved in chronic obstructive pulmonary disease (COPD) exacerbations. Cigarette smoke is the best known risk factor for COPD. Health concerns of microbes in tobacco are starting to be acknowledged. Other groups showed that tobacco flakes inhaled from cigarettes could carry bacteria into the lungs. Moreover, bacterial products such as LPS remain in cigarette smoke and could contribute to inflammation. Our group is one of the first suggesting that Toll-like receptors (TLRs) are involved in the pathogenesis of COPD. TLRs are activated by microbial products. Our in vitro studies suggest that cigarette smoke induces inflammation partly via TLR9. TLR9 is activated by bacterial DNA. During the Aspen Lung Conference we provided further evidence for a role for TLR9 in the development of COPD. First, we demonstrated that chronic (5 weeks) activation of pulmonary TLR9 in mice leads to lung inflammation, emphysema, heart hypertrophy, and reduced airway function. Moreover, novel data were presented showing that not all bacteria are destroyed during tobacco burning. When smoke from burning cigarettes is passed over agar plates, colony-forming units clearly follow smoke deposition patterns unrelated to tobacco flake transmission. Currently, a role for bioaerosol exposure in the development of COPD is still underappreciated. Our data emphasize that certain microorganisms and their products could become airborne during combustion. This information is relevant for (passive) smokers but also for women and children who use biomass for cooking indoors in developing countries. Better insight into cause
S76
1
Division of Pulmonary Sciences and Critical Care Medicine, and 2Department of Pediatrics, Pulmonary Medicine, University of Colorado Denver, Denver, Colorado
Background: Bacterial infection of lung airways underlies some of the main complications of chronic obstructive lung disease (COPD), significantly impacting disease progression and outcome. Colonization by bacteria may synergize, amplify, or trigger pathways of tissue damage started by cigarette smoke, leading to airway inflammation with remodeling and alveolar destruction. Elucidation of the presence and types of lung bacterial populations in different stages of COPD may reveal important insights into the pathobiology of the disease. Hypothesis: Sequencing of the bacterial small subunit ribosomal RNA gene in 55 well-characterized clinical lung samples (5 normal, 4 G0, 7 G1-2, 27 G3-G4, and 12 patients with cystic fibrosis) revealed the presence of Novosphingobium spp. (. 2% abundance) in G3-G4 COPD, CF, and control subjects. We sought to quantify levels of Novosphingobium spp. in clinical samples and to investigate the role of Novosphingobium spp. in a subacute inflammation mouse model. Methods: Freshly frozen human lung samples were analyzed for levels of Novosphingobium ssp. by quantitative polymerase chain reaction. C57BL/6 mice were challenged intratracheally with Novosphingobium panipatense or phosphate-buffered saline and exposed to cigarette smoke or room air for 6 weeks. Bronchoalveolar lavage was harvested and evaluated for inflammation markers. Lung tissue was analyzed by flow cytometry for markers of inflammation. Results: Quantitative polymerase chain reaction was concordant with prior sequence data, and high levels of Novosphingobium AnnalsATS Volume 11 Supplement 1 | January 2014
ABSTRACTS were quantifiable in advanced COPD but not from other disease stages. Bronchoalveolar lavage neutrophil and macrophage counts were significantly higher in mice challenged intratracheally with N. panipatense compared with control mice (P , 0.01). Frequencies of neutrophils and macrophages in lung tissue were increased in mice challenged with N. panipatense in room air compared with control mice. The effect of cigarette smoke in conjunction with N. panipatense was unclear and necessitates further elucidation. Conclusions: Novosphingobium ssp. was associated with lung inflammation in mice and may play a role in more severe COPD disease. Author disclosures are available with the text of this abstract at www.atsjournals.org.
(Received in original form June 27, 2013; accepted in final form July 1, 2013) Correspondence and requests for reprints should be addressed to J. Kirk Harris, Ph.D., Department of Pediatrics, Pulmonary Medicine, University of Colorado Denver, 13123 E. 16th Avenue, Box B395, Aurora, CO 80045. E-mail:
[email protected] Ann Am Thorac Soc Vol 11, Supplement 1, pp S76–S77, Jan 2014 Copyright © 2014 by the American Thoracic Society Internet address: www.atsjournals.org
from lung apex to base (McDonough JE, et al., N Engl J Med 2011;365:1567–1576). From each slice one sample was examined by micro–computed tomography to measure emphysematous destruction, another to measure the inflammatory immune cell infiltration by quantitative histology, and a third was used to study bacteria by quantitative polymerase chain reaction and 454 pyrotag sequencing (Sze MA, et al., Am J Respir Crit Care Med 2012;185:1073–1080). Results: The average numbers of 16S/1000Rpp40 gene copies were 7.44 6 18.6 and 7.77 6 16.3 (COPD vs. controls; P . 0.05). In contrast, comparison of upper lung tissue samples (COPD vs. controls) showed an increase in detectable bacteria (P , 0.05) with no difference between lower lung tissue samples (P . 0.05). Sequencing identified a core population of bacteria that was significantly associated (P , 0.05) with the inflammatory and tissue-remodeling response as well as with surface area and numbers of terminal bronchioles (P , 0.05). Conclusion: The accumulations of specific bacteria within the human lung microbiome are associated with inflammatory cell infiltration and progressive emphysematous destruction. Author disclosures are available with the text of this abstract at www.atsjournals.org.
Received in original form June 24, 2013; accepted in final form June 24, 2013 Supported by Merck IIS 38970, NIH # 5P50HL084922, 5P50HL084948, 1R01HL95388, and CIHR # CIF-97687.
Host Response to the Lung Microbiome in Lung Tissue Undergoing Emphysematous Destruction Marc Sze1, Pedro A. Dimitriu2, Masaru Suzuki1, John E. McDonough1, John V. Gosselink1, Mark W. Elliott1, William W. Mohn2, Don D. Sin1, Shizu Hayashi1, and James C. Hogg1 1 UBC James Hogg Research Centre, St. Paul’s Hospital/ Providence Health Care-University of British Columbia, and 2Department of Immunology and Microbiology, University of British Columbia, Vancouver, British Columbia, Canada
Correspondence and requests for reprints should be addressed to Marc Sze, UBC James Hogg Research Centre, Room 166, 1081 Burrard Street, Vancouver, BC, Canada V6Z 1Y6. E-mail:
[email protected] Ann Am Thorac Soc Vol 11, Supplement 1, p S77, Jan 2014 Copyright © 2014 by the American Thoracic Society Internet address: www.atsjournals.org
The Lung Microbiome in Moderate and Severe Chronic Obstructive Pulmonary Disease Alexa A. Pragman1, Hyeun Bum Kim2, Cavan S. Reilly3, Christine Wendt4, and Richard E. Isaacson2 1
Rationale: The progression of chronic obstructive pulmonary disease (COPD) has been associated with infiltration of peripheral lung tissue by inflammatory immune cells that form tertiary lymphoid organs. These observations have documented the presence of an adaptive immune response without indentifying the antigens that drive it (Hogg JC, et al., N Engl J Med 2004;350:2645– 2653).
Department of Medicine, Division of Infectious Diseases and International Medicine, University of Minnesota Medical Center, St. Paul, Minnesota; and Departments of 2Veterinary 4 3 Biosciences, Biostatistics, and Medicine, University of Minnesota, St. Paul, Minnesota
Hypothesis: Bacteria within the human lung microbiome drive the host inflammatory immune response associated with progression of COPD.
Chronic obstructive pulmonary disease (COPD) is an inflammatory disorder characterized by incompletely reversible airflow obstruction. Bacterial lung infection contributes to roughly 50% of COPD exacerbations. Even during periods of stable lung function, the lung harbors a community of bacteria, termed the microbiome. The role of the lung microbiome in the pathogenesis of COPD remains unknown. The COPD lung microbiome, like the healthy lung microbiome, appears to reflect microaspiration of oral microflora.
Methods: Whole lungs donated by patients with severe COPD (n = 5) were compared with unused donor lungs (n = 4) that served as controls. These specimens were inflated with air, rapidly frozen in liquid nitrogen vapor, and cut into 2-cm-thick slices
Abstracts
S77
