Skip to main content
Top
Published in: Respiratory Research 1/2018

Open Access 01-12-2018 | Research

Sequential broncho-alveolar lavages reflect distinct pulmonary compartments: clinical and research implications in lung transplantation

Authors: Liran Levy, Stephen C. Juvet, Kristen Boonstra, Lianne G. Singer, Sassan Azad, Betty Joe, Marcelo Cypel, Shaf Keshavjee, Tereza Martinu

Published in: Respiratory Research | Issue 1/2018

Login to get access

Abstract

Background

Bronchoalveolar lavage (BAL) has proven to be very useful to monitor the lung allograft after transplantation. In addition to allowing detection of infections, multiple BAL analytes have been proposed as potential biomarkers of lung allograft rejection or dysfunction. However, BAL collection is not well standardized and differences in BAL collection represent an important source of variation. We hypothesized that there are systematic differences between sequential BALs that are relevant to BAL analysis.

Methods

As part of 126 consecutive bronchoscopies in lung transplant recipients, two sequential BALs (BAL1 and BAL2) were performed in one location during each bronchoscopy by instilling and suctioning 50 ml of normal saline twice into separate containers. Cell concentration, viability and differentials, Surfactant Protein-D (SP-D), Club Cell Secretory Protein (CCSP), and levels of CXCL10, IL-10, CCL2, CCL5, VEGF-C, RAGE, CXCL9, CXCL1, IL-17A, IL-21, PDGF, and GCSF were compared between BAL1 and BAL2.

Results

Total cell concentration did not differ between BAL1 and BAL2; however, compared to BAL2, BAL1 had more dead cells, epithelial cells, neutrophils, and higher concentrations of airway epithelium-derived CCSP and inflammatory markers. BAL2 had a higher concentration of SP-D compared to BAL1.

Conclusion

In this study performed in lung transplant recipients, we show that sequential BALs represent different lung compartments and have distinct compositions. BAL1 represents the airway compartment with more epithelial cells, neutrophils, and epithelium-derived CCSP. Conversely, BAL2 samples preferentially the distal bronchoalveolar space with greater cell viability and higher SP-D. Our findings illustrate how the method of BAL collection can influence analyte concentrations and further emphasize the need for a standardized approach in translational research involving BAL samples.
Appendix
Available only for authorised users
Literature
1.
go back to reference Guilinger RA, Paradis IL, Dauber JH, Yousem SA, Williams PA, Keenan RJ, Griffith BP. The importance of bronchoscopy with transbronchial biopsy and bronchoalveolar lavage in the management of lung transplant recipients. Am J Respir Crit Care Med. 1995;152:2037–43.CrossRefPubMed Guilinger RA, Paradis IL, Dauber JH, Yousem SA, Williams PA, Keenan RJ, Griffith BP. The importance of bronchoscopy with transbronchial biopsy and bronchoalveolar lavage in the management of lung transplant recipients. Am J Respir Crit Care Med. 1995;152:2037–43.CrossRefPubMed
2.
go back to reference Haslam PL, Baughman RP. Report of ERS task force: guidelines for measurement of acellular components and standardization of BAL. Eur Respir J. 1999;14:245–8.CrossRefPubMed Haslam PL, Baughman RP. Report of ERS task force: guidelines for measurement of acellular components and standardization of BAL. Eur Respir J. 1999;14:245–8.CrossRefPubMed
3.
go back to reference Husain S, Resende MR, Rajwans N, Zamel R, Pilewski JM, Crespo MM, Singer LG, McCurry KR, Kolls JK, Keshavjee S, Liles WC. Elevated CXCL10 (IP-10) in bronchoalveolar lavage fluid is associated with acute cellular rejection after human lung transplantation. Transplantation. 2014;97:90–7. Husain S, Resende MR, Rajwans N, Zamel R, Pilewski JM, Crespo MM, Singer LG, McCurry KR, Kolls JK, Keshavjee S, Liles WC. Elevated CXCL10 (IP-10) in bronchoalveolar lavage fluid is associated with acute cellular rejection after human lung transplantation. Transplantation. 2014;97:90–7.
4.
go back to reference Madsen CB, Norgaard A, Iversen M, Ryder LP. Elevated mRNA levels of CTLA-4, FoxP3, and granzyme B in BAL, but not in blood, during acute rejection of lung allografts. Transpl Immunol. 2010;24:26–32.CrossRefPubMed Madsen CB, Norgaard A, Iversen M, Ryder LP. Elevated mRNA levels of CTLA-4, FoxP3, and granzyme B in BAL, but not in blood, during acute rejection of lung allografts. Transpl Immunol. 2010;24:26–32.CrossRefPubMed
5.
go back to reference Vos R, Vanaudenaerde BM, De Vleeschauwer SI, Willems-Widyastuti A, Dupont LJ, Van Raemdonck DE, Verleden GM. C-reactive protein in bronchoalveolar lavage fluid is associated with markers of airway inflammation after lung transplantation. Transplant Proc. 2009;41:3409–13.CrossRefPubMed Vos R, Vanaudenaerde BM, De Vleeschauwer SI, Willems-Widyastuti A, Dupont LJ, Van Raemdonck DE, Verleden GM. C-reactive protein in bronchoalveolar lavage fluid is associated with markers of airway inflammation after lung transplantation. Transplant Proc. 2009;41:3409–13.CrossRefPubMed
6.
go back to reference Gimino VJ, Lande JD, Berryman TR, King RA, Hertz MI. Gene expression profiling of bronchoalveolar lavage cells in acute lung rejection. Am J Respir Crit Care Med. 2003;168:1237–42.CrossRefPubMed Gimino VJ, Lande JD, Berryman TR, King RA, Hertz MI. Gene expression profiling of bronchoalveolar lavage cells in acute lung rejection. Am J Respir Crit Care Med. 2003;168:1237–42.CrossRefPubMed
7.
go back to reference Kennedy VE, Todd JL, Palmer SM. Bronchoalveolar lavage as a tool to predict, diagnose and understand bronchiolitis obliterans syndrome. Am J Transplant. 2013;13:552–61.CrossRefPubMedPubMedCentral Kennedy VE, Todd JL, Palmer SM. Bronchoalveolar lavage as a tool to predict, diagnose and understand bronchiolitis obliterans syndrome. Am J Transplant. 2013;13:552–61.CrossRefPubMedPubMedCentral
8.
go back to reference Benjamin-Alexander Bollmann BS, Drick N, Welte T, Gottlieb JT, Greer M. Cellular analysis in bronchoalveolar lavage: inherent limitations of current standard procedure. Eur Respir J. 2017;49:1601844. Benjamin-Alexander Bollmann BS, Drick N, Welte T, Gottlieb JT, Greer M. Cellular analysis in bronchoalveolar lavage: inherent limitations of current standard procedure. Eur Respir J. 2017;49:1601844.
9.
go back to reference Meyer KC, Raghu G, Baughman RP, Brown KK, Costabel U, du Bois RM, Drent M, Haslam PL, Kim DS, Nagai S, et al. An official American Thoracic Society clinical practice guideline: the clinical utility of bronchoalveolar lavage cellular analysis in interstitial lung disease. Am J Respir Crit Care Med. 2012;185:1004–14.CrossRefPubMed Meyer KC, Raghu G, Baughman RP, Brown KK, Costabel U, du Bois RM, Drent M, Haslam PL, Kim DS, Nagai S, et al. An official American Thoracic Society clinical practice guideline: the clinical utility of bronchoalveolar lavage cellular analysis in interstitial lung disease. Am J Respir Crit Care Med. 2012;185:1004–14.CrossRefPubMed
10.
go back to reference Speck NE, Schuurmans MM, Murer C, Benden C, Huber LC. Diagnostic value of plasma and bronchoalveolar lavage samples in acute lung allograft rejection: differential cytology. Respir Res. 2016;17:74.CrossRefPubMedPubMedCentral Speck NE, Schuurmans MM, Murer C, Benden C, Huber LC. Diagnostic value of plasma and bronchoalveolar lavage samples in acute lung allograft rejection: differential cytology. Respir Res. 2016;17:74.CrossRefPubMedPubMedCentral
11.
go back to reference D'Ovidio F, Mura M, Ridsdale R, Takahashi H, Waddell TK, Hutcheon M, Hadjiliadis D, Singer LG, Pierre A, Chaparro C, et al. The effect of reflux and bile acid aspiration on the lung allograft and its surfactant and innate immunity molecules SP-A and SP-D. Am J Transplant. 2006;6:1930–8.CrossRefPubMed D'Ovidio F, Mura M, Ridsdale R, Takahashi H, Waddell TK, Hutcheon M, Hadjiliadis D, Singer LG, Pierre A, Chaparro C, et al. The effect of reflux and bile acid aspiration on the lung allograft and its surfactant and innate immunity molecules SP-A and SP-D. Am J Transplant. 2006;6:1930–8.CrossRefPubMed
12.
go back to reference Nord M, Schubert K, Cassel TN, Andersson O, Riise GC. Decreased serum and bronchoalveolar lavage levels of Clara cell secretory protein (CC16) is associated with bronchiolitis obliterans syndrome and airway neutrophilia in lung transplant recipients. Transplantation. 2002;73:1264–9.CrossRefPubMed Nord M, Schubert K, Cassel TN, Andersson O, Riise GC. Decreased serum and bronchoalveolar lavage levels of Clara cell secretory protein (CC16) is associated with bronchiolitis obliterans syndrome and airway neutrophilia in lung transplant recipients. Transplantation. 2002;73:1264–9.CrossRefPubMed
13.
go back to reference Kelly FL, Kennedy VE, Jain R, Sindhwani NS, Finlen Copeland CA, Snyder LD, Eu JP, Meltzer EB, Brockway BL, Pavlisko E, et al. Epithelial clara cell injury occurs in bronchiolitis obliterans syndrome after human lung transplantation. Am J Transplant. 2012;12:3076–84.CrossRefPubMedPubMedCentral Kelly FL, Kennedy VE, Jain R, Sindhwani NS, Finlen Copeland CA, Snyder LD, Eu JP, Meltzer EB, Brockway BL, Pavlisko E, et al. Epithelial clara cell injury occurs in bronchiolitis obliterans syndrome after human lung transplantation. Am J Transplant. 2012;12:3076–84.CrossRefPubMedPubMedCentral
14.
go back to reference Patella M, Anile M, Del Porto P, Diso D, Pecoraro Y, Onorati I, Mantovani S, De Giacomo T, Ascenzioni F, Rendina EA, Venuta F. Role of cytokine profile in the differential diagnosis between acute lung rejection and pulmonary infections after lung transplantationdagger. Eur J Cardiothorac Surg. 2015;47:1031–6.CrossRefPubMed Patella M, Anile M, Del Porto P, Diso D, Pecoraro Y, Onorati I, Mantovani S, De Giacomo T, Ascenzioni F, Rendina EA, Venuta F. Role of cytokine profile in the differential diagnosis between acute lung rejection and pulmonary infections after lung transplantationdagger. Eur J Cardiothorac Surg. 2015;47:1031–6.CrossRefPubMed
15.
go back to reference Verleden SE, Ruttens D, Vos R, Vandermeulen E, Moelants E, Mortier A, Van Raemdonck DE, Proost P, Schols D, Verleden GM, Vanaudenaerde BM. Differential cytokine, chemokine and growth factor expression in phenotypes of chronic lung allograft dysfunction. Transplantation. 2015;99:86–93.CrossRefPubMed Verleden SE, Ruttens D, Vos R, Vandermeulen E, Moelants E, Mortier A, Van Raemdonck DE, Proost P, Schols D, Verleden GM, Vanaudenaerde BM. Differential cytokine, chemokine and growth factor expression in phenotypes of chronic lung allograft dysfunction. Transplantation. 2015;99:86–93.CrossRefPubMed
16.
go back to reference Pelaez A, Force SD, Gal AA, Neujahr DC, Ramirez AM, Naik PM, Quintero DA, Pileggi AV, Easley KA, Echeverry R, et al. Receptor for advanced glycation end products in donor lungs is associated with primary graft dysfunction after lung transplantation. Am J Transplant. 2010;10:900–7.CrossRefPubMedPubMedCentral Pelaez A, Force SD, Gal AA, Neujahr DC, Ramirez AM, Naik PM, Quintero DA, Pileggi AV, Easley KA, Echeverry R, et al. Receptor for advanced glycation end products in donor lungs is associated with primary graft dysfunction after lung transplantation. Am J Transplant. 2010;10:900–7.CrossRefPubMedPubMedCentral
17.
go back to reference Belperio JA, Keane MP, Burdick MD, Gomperts BN, Xue YY, Hong K, Mestas J, Zisman D, Ardehali A, Saggar R, et al. CXCR2/CXCR2 ligand biology during lung transplant ischemia-reperfusion injury. J Immunol. 2005;175:6931–9.CrossRefPubMed Belperio JA, Keane MP, Burdick MD, Gomperts BN, Xue YY, Hong K, Mestas J, Zisman D, Ardehali A, Saggar R, et al. CXCR2/CXCR2 ligand biology during lung transplant ischemia-reperfusion injury. J Immunol. 2005;175:6931–9.CrossRefPubMed
18.
go back to reference Vanaudenaerde BM, De Vleeschauwer SI, Vos R, Meyts I, Bullens DM, Reynders V, Wuyts WA, Van Raemdonck DE, Dupont LJ, Verleden GM. The role of the IL23/IL17 axis in bronchiolitis obliterans syndrome after lung transplantation. Am J Transplant. 2008;8:1911–20.CrossRefPubMed Vanaudenaerde BM, De Vleeschauwer SI, Vos R, Meyts I, Bullens DM, Reynders V, Wuyts WA, Van Raemdonck DE, Dupont LJ, Verleden GM. The role of the IL23/IL17 axis in bronchiolitis obliterans syndrome after lung transplantation. Am J Transplant. 2008;8:1911–20.CrossRefPubMed
20.
go back to reference Hertz MI, Henke CA, Nakhleh RE, Harmon KR, Marinelli WA, Fox JM, Kubo SH, Shumway SJ, Bolman RM 3rd, Bitterman PB. Obliterative bronchiolitis after lung transplantation: a fibroproliferative disorder associated with platelet-derived growth factor. Proc Natl Acad Sci U S A. 1992;89:10385–9.CrossRefPubMedPubMedCentral Hertz MI, Henke CA, Nakhleh RE, Harmon KR, Marinelli WA, Fox JM, Kubo SH, Shumway SJ, Bolman RM 3rd, Bitterman PB. Obliterative bronchiolitis after lung transplantation: a fibroproliferative disorder associated with platelet-derived growth factor. Proc Natl Acad Sci U S A. 1992;89:10385–9.CrossRefPubMedPubMedCentral
21.
go back to reference Bergmann M, Tiroke A, Schafer H, Barth J, Haverich A. Gene expression of profibrotic mediators in bronchiolitis obliterans syndrome after lung transplantation. Scand Cardiovasc J. 1998;32:97–103.CrossRefPubMed Bergmann M, Tiroke A, Schafer H, Barth J, Haverich A. Gene expression of profibrotic mediators in bronchiolitis obliterans syndrome after lung transplantation. Scand Cardiovasc J. 1998;32:97–103.CrossRefPubMed
22.
go back to reference Verleden SE, Vos R, Mertens V, Willems-Widyastuti A, De Vleeschauwer SI, Dupont LJ, Verleden GM, Van Raemdonck DE, Vanaudenaerde BM. Heterogeneity of chronic lung allograft dysfunction: insights from protein expression in broncho alveolar lavage. J Heart Lung Transplant. 2011;30:667–73.CrossRefPubMed Verleden SE, Vos R, Mertens V, Willems-Widyastuti A, De Vleeschauwer SI, Dupont LJ, Verleden GM, Van Raemdonck DE, Vanaudenaerde BM. Heterogeneity of chronic lung allograft dysfunction: insights from protein expression in broncho alveolar lavage. J Heart Lung Transplant. 2011;30:667–73.CrossRefPubMed
23.
go back to reference Dosanjh A, Robbins RC. Cytokines interleukin 5 and GM-CSF in the BAL fluid of lung transplant patients. Chest. 1998;114:349.CrossRefPubMed Dosanjh A, Robbins RC. Cytokines interleukin 5 and GM-CSF in the BAL fluid of lung transplant patients. Chest. 1998;114:349.CrossRefPubMed
24.
go back to reference Tinckam KJ, Keshavjee S, Chaparro C, Barth D, Azad S, Binnie M, Chow CW, de Perrot M, Pierre AF, Waddell TK, et al. Survival in sensitized lung transplant recipients with perioperative desensitization. Am J Transplant. 2015;15:417–26.CrossRefPubMed Tinckam KJ, Keshavjee S, Chaparro C, Barth D, Azad S, Binnie M, Chow CW, de Perrot M, Pierre AF, Waddell TK, et al. Survival in sensitized lung transplant recipients with perioperative desensitization. Am J Transplant. 2015;15:417–26.CrossRefPubMed
25.
go back to reference Gutierrez C, Al-Faifi S, Chaparro C, Waddell T, Hadjiliadis D, Singer L, Keshavjee S, Hutcheon M. The effect of recipient's age on lung transplant outcome. Am J Transplant. 2007;7:1271–7.CrossRefPubMed Gutierrez C, Al-Faifi S, Chaparro C, Waddell T, Hadjiliadis D, Singer L, Keshavjee S, Hutcheon M. The effect of recipient's age on lung transplant outcome. Am J Transplant. 2007;7:1271–7.CrossRefPubMed
26.
go back to reference Verleden GM, Raghu G, Meyer KC, Glanville AR, Corris P. A new classification system for chronic lung allograft dysfunction. J Heart Lung Transplant. 2014;33:127–33.CrossRefPubMed Verleden GM, Raghu G, Meyer KC, Glanville AR, Corris P. A new classification system for chronic lung allograft dysfunction. J Heart Lung Transplant. 2014;33:127–33.CrossRefPubMed
27.
go back to reference Vanaudenaerde BM, Wuyts WA, Geudens N, Nawrot TS, Vos R, Dupont LJ, Van Raemdonck DE, Verleden GM. Broncho-alveolar lavage fluid recovery correlates with airway neutrophilia in lung transplant patients. Respir Med. 2008;102:339–47.CrossRefPubMed Vanaudenaerde BM, Wuyts WA, Geudens N, Nawrot TS, Vos R, Dupont LJ, Van Raemdonck DE, Verleden GM. Broncho-alveolar lavage fluid recovery correlates with airway neutrophilia in lung transplant patients. Respir Med. 2008;102:339–47.CrossRefPubMed
28.
go back to reference Kelly CA, Kotre CJ, Ward C, Hendrick DJ, Walters EH. Anatomical distribution of bronchoalveolar lavage fluid as assessed by digital subtraction radiography. Thorax. 1987;42:624–8.CrossRefPubMedPubMedCentral Kelly CA, Kotre CJ, Ward C, Hendrick DJ, Walters EH. Anatomical distribution of bronchoalveolar lavage fluid as assessed by digital subtraction radiography. Thorax. 1987;42:624–8.CrossRefPubMedPubMedCentral
29.
30.
go back to reference Davis GS, Giancola MS, Costanza MC, Low RB. Analyses of sequential bronchoalveolar lavage samples from healthy human volunteers. Am Rev Respir Dis. 1982;126:611–6.PubMed Davis GS, Giancola MS, Costanza MC, Low RB. Analyses of sequential bronchoalveolar lavage samples from healthy human volunteers. Am Rev Respir Dis. 1982;126:611–6.PubMed
31.
go back to reference Lam S, Leriche JC, Kijek K, Phillips D. Effect of bronchial lavage volume on cellular and protein recovery. Chest. 1985;88:856–9.CrossRefPubMed Lam S, Leriche JC, Kijek K, Phillips D. Effect of bronchial lavage volume on cellular and protein recovery. Chest. 1985;88:856–9.CrossRefPubMed
32.
go back to reference Merrill W, O'Hearn E, Rankin J, Naegel G, Matthay RA, Reynolds HY. Kinetic analysis of respiratory tract proteins recovered during a sequential lavage protocol. Am Rev Respir Dis. 1982;126:617–20.PubMed Merrill W, O'Hearn E, Rankin J, Naegel G, Matthay RA, Reynolds HY. Kinetic analysis of respiratory tract proteins recovered during a sequential lavage protocol. Am Rev Respir Dis. 1982;126:617–20.PubMed
33.
go back to reference Rennard SI, Ghafouri M, Thompson AB, Linder J, Vaughan W, Jones K, Ertl RF, Christensen K, Prince A, Stahl MG, et al. Fractional processing of sequential bronchoalveolar lavage to separate bronchial and alveolar samples. Am Rev Respir Dis. 1990;141:208–17.CrossRefPubMed Rennard SI, Ghafouri M, Thompson AB, Linder J, Vaughan W, Jones K, Ertl RF, Christensen K, Prince A, Stahl MG, et al. Fractional processing of sequential bronchoalveolar lavage to separate bronchial and alveolar samples. Am Rev Respir Dis. 1990;141:208–17.CrossRefPubMed
34.
go back to reference Yasuoka S, Nakayama T, Kawano T, Ogushi F, Doi H, Hayashi H, Tsubura E. Comparison of cell profiles of bronchial and bronchoalveolar lavage fluids between normal subjects and patients with idiopathic pulmonary fibrosis. Tohoku J Exp Med. 1985;146:33–45.CrossRefPubMed Yasuoka S, Nakayama T, Kawano T, Ogushi F, Doi H, Hayashi H, Tsubura E. Comparison of cell profiles of bronchial and bronchoalveolar lavage fluids between normal subjects and patients with idiopathic pulmonary fibrosis. Tohoku J Exp Med. 1985;146:33–45.CrossRefPubMed
35.
go back to reference Baughman RP. Technical aspects of bronchoalveolar lavage: recommendations for a standard procedure. Semin Respir Crit Care Med. 2007;28:475–85.CrossRefPubMed Baughman RP. Technical aspects of bronchoalveolar lavage: recommendations for a standard procedure. Semin Respir Crit Care Med. 2007;28:475–85.CrossRefPubMed
36.
go back to reference Jones KP, Edwards JH, Reynolds SP, Peters TJ, Davies BH. A comparison of albumin and urea as reference markers in bronchoalveolar lavage fluid from patients with interstitial lung disease. Eur Respir J. 1990;3:152–6.PubMed Jones KP, Edwards JH, Reynolds SP, Peters TJ, Davies BH. A comparison of albumin and urea as reference markers in bronchoalveolar lavage fluid from patients with interstitial lung disease. Eur Respir J. 1990;3:152–6.PubMed
37.
go back to reference Ward C, Fenwick J, Booth H, Walters EH. Albumin is not suitable as a marker of bronchoalveolar lavage dilution in interstitial lung disease. Eur Respir J. 1997;10:2029–33.CrossRefPubMed Ward C, Fenwick J, Booth H, Walters EH. Albumin is not suitable as a marker of bronchoalveolar lavage dilution in interstitial lung disease. Eur Respir J. 1997;10:2029–33.CrossRefPubMed
Metadata
Title
Sequential broncho-alveolar lavages reflect distinct pulmonary compartments: clinical and research implications in lung transplantation
Authors
Liran Levy
Stephen C. Juvet
Kristen Boonstra
Lianne G. Singer
Sassan Azad
Betty Joe
Marcelo Cypel
Shaf Keshavjee
Tereza Martinu
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Respiratory Research / Issue 1/2018
Electronic ISSN: 1465-993X
DOI
https://doi.org/10.1186/s12931-018-0786-z

Other articles of this Issue 1/2018

Respiratory Research 1/2018 Go to the issue