Use of Bronchoalveolar Lavage to Detect Lung Injury

Rogene F. Henderson1, Bruce A. Muggenburg1

1 Lovelace Respiratory Research Institute, Albuquerque, New Mexico
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 18.4
DOI:  10.1002/0471140856.tx1804s21
Online Posting Date:  September, 2004
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Abstract

This unit describes how bronchoalveolar lavage can be used in laboratory animals to sample the epithelial lining fluid of the lung for information on the degree of pulmonary inflammation induced by exposure to an airborne toxicant. The technique allows quantitative assessment of inflammatory responses and is valuable for providing doseā€response information in exposed animals. Lavage fluid samples may also be used for proteomic analyses, and the protein expression profiles may be used to address specific mechanistic questions.

Keywords: lung; inhalation toxicity; epithelial lining fluid; bronchoalveolar lavage; pulmonary toxicity

     
 
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Table of Contents

  • Strategic Planning
  • Basic Protocol 1: Bronchoalveolar Lavage Performed in Large Animals In Vivo
  • Alternate Protocol 1: Segmental Lavage
  • Alternate Protocol 2: Bronchoalveolar Lavage Performed in the Excised Lung
  • Support Protocol 1: Processing Bronchoalveolar Lavage Fluid (BALF) and Analysis of Lung Damage Indicators
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Bronchoalveolar Lavage Performed in Large Animals In Vivo

  Materials
  • Animal: dog or other large animal
  • Preanesthesia agents (e.g., acepromazine, ketamine, zylazine)
  • Atropine
  • General anesthetic: 5% (for induction) and 2% (for maintenance) isoflurane in O 2
  • Sterile wash fluid (see )
  • Face mask
  • Endotracheal tube with balloon (Rusch)
  • Whole lavage apparatus (see )
  • Additional reagents and equipment for storing and processing BALF fluid (see protocol 4)

Alternate Protocol 1: Segmental Lavage

  • Fiberoptic bronchoscope (e.g., Olympus, Pentex)
  • Syringes

Alternate Protocol 2: Bronchoalveolar Lavage Performed in the Excised Lung

  • Anesthetic for euthanasia (e.g., 5% halothane in O 2, sodium pentobarbital)
  • Dissection board and tools
  • Plastic tracheal catheter with Luer lock connector
  • Syringes

Support Protocol 1: Processing Bronchoalveolar Lavage Fluid (BALF) and Analysis of Lung Damage Indicators

  • BALF (see protocol 1 or Alternate Protocols protocol 21 or protocol 32)
  • Additional reagents and equipment for counting cells ( appendix 3B)
NOTE: Keep BALF on ice at all times.
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Figures

Videos

Literature Cited

   Bergmeyer, H.U. (ed.) 1983. Methods of Enzymatic Analysis, 3rd edition, VCH Publishing, Weinheim, Germany.
   Henderson, R.F. 1988. Use of bronchoalveolar lavage to detect lung damage. In Toxicology of the Lung (D.E. Gardner, J.D. Crapo, and E. J. Massaro, eds.) pp. 239‐268. Raven Press, New York.
   Henderson, R.F. 1989. Bronchoalveolar lavage: A tool for assessing the health status of the lung. In Concepts in Inhalation Toxicology (R.O. McClellan and R.F. Henderson, eds.) pp. 415‐444. Hemisphere Publishing, New York.
   Henderson, R.F. and Lowry, J.S. 1983. Effect of anesthetic agents on lavage fluid parameters used as indicators of pulmonary injury. Lab. Anim. Sci. 33:60‐62.
   Henderson, R.F. and Muggenburg, B.A. 1992. Bronchoalveolar lavage in animals. In Bronchoalveolar Lavage (R.P. Baughman, ed.) pp. 265‐287. Mosby‐Year Book, St. Louis.
   Mauderly, J.L. 1977. Bronchopulmonary lavage of small laboratory animals. Lab. Anim. Sci. 27:255‐261.
   Rennard, S.I., Albers, R., Bleecker, E., Klech, H., Rosenwasser, L., Olivieri, D., and Sibelle, Y. 1998. Bronchoalveolar lavage: Performance, sampling procedure, processing and assessment. Eur. Respir. J. Suppl. 26:13S‐15S.
   Strober, W. 1997. Wright‐Giemsa and nonspecific esterase staining of cells. In Current Protocols in Immunology (J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, and W. Strober, eds.) pp. A.3C.1‐A.3C.3. John Wiley & Sons, Hoboken, N.J.
Key References
   Henderson, 1989. See above.
  Reviews basic principles of the use of BALF analyses to quantitate lung responses in animals.
   Rennard et al., 1998. See above.
  Useful for comparing animal techniques with those used in human clinical studies.
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