Bronchoalveolar Lavage (Bal)

Bronchoalveolar Lavage (BAL) Technique

Going beyond respiratory mechanics

In a pre-clinical setting, respiratory mechanics measurements provide in-depth understanding into the lung’s function, structures and volumes. Complementary techniques are often performed to provide even further insight into factors leading to disease states and progression. A common procedure performed in murine models following lung function measurements is the Bronchoalveolar Lavage (BAL).

The bronchoalveolar lavage (BAL) technique

Bronchoalveolar lavage is a technique used both in clinical and preclinical settings. It allows for sampling of the cellular and molecular composition of the airways, making it particularly useful to monitor the inflammatory and immune responses. BALs are performed by injecting a stable buffer solution (such as PBS or saline) into the bronchioles and then gently retrieving it through aspiration in order to obtain what is called the BALF: bronchoalveolar lavage fluid. BALs are a quick, inexpensive and highly reproducible technique and can be easily performed right after the end of lung function measurements with the flexiVent system, as the subject is already tracheostomized, cannulated and ready for the injection.

Analyzing bronchoalveolar lavage fluid (BALF)

Retrieving the cellular and non-cellular content of the epithelial lining fluid of the lungs allows for a quantitative and qualitative analysis of such content through various methods including ELISA, immunoblot, Flow Cytometry, immunohistochemistry, PCR, HPLC and more.

Bronchoalveolar lavage fluid (BALF) can contain antibodies and cytokines involved in the attraction of cells implicated in the immune and inflammatory responses of disease models such as asthma. Evaluating the level of these proteins in BALF can provide insight into altered cellular influx and each cell’s implication in airway hyperresponsiveness (AHR) and disease development. This can help find new therapeutic targets and pathways for disease prevention and treatment, as well as evaluate drug efficacy.

This technique can also enumerate and identify cell subtypes in the BAL fluid, often to quantify levels of inflammatory cells such as eosinophils, neutrophils and lymphocytes to evaluate cellular infiltration of the airways (Chesne et al.).

flexiVent measurements followed by BAL

There are many published studies where BALF is collected following respiratory mechanics measurements with the flexiVent FX system. Below are a few recent publications among the extensive list available in the literature:

  • Julie Chesne et al. (2014). Prime role of IL-17A in neutrophilia and airway smooth muscle contraction in a house dust mite–induced allergic asthma model. Volume 135, Issue 6, Pages 1643–1645.e5. Doi: https://doi.org/10.1016/j.jaci.2014.12.1872
  • Davis, I.C., et al. (2019). Treatment with Liponucleotides Attenuates DRA-Induced Asthma in Mice. American Journal of Respiratory and Critical Care Medicine;199: A3827
  • Ramos Ramirez, P., et al. (2019). Budesonide-induced decrease of IL-5 production does not affect allergen-induced bronchoconstriction or AHR in a novel guinea pig asthma model. European Respiratory Journal, 54: PA4081; DOI: 10.1183/13993003.congress-2019.PA4081
  • Mariappan, N., et al. (2018). Pulmonary Manifestations of Inhaled Arsenic Trioxide Following an Acute Accidental Exposure. American Journal of Respiratory and Critical Care Medicine; 197: A2974
  • Woo, L., et al. (2018). Treatment with a CD200 Receptor Agonist Aptamer Attenuates Airway Hyperresponsiveness, Airway Inflammation, and Remodelling in a Chronic Murine Model of HDM-Induced Airways Inflammation. American Journal of Respiratory and Critical Care Medicine ;197: A1415
  • Hamakawa, Y., et al. (2018). Muscarinic Receptor Stimulation Enhances Pathophysiological Phenotypes in Murine Model of COPD. American Journal of Respiratory and Critical Care Medicine 2018;197: A7158

The Journal of Visualized Experiments (JoVE) recently published a comprehensive guide on performing Bronchoalveolar Lavage in mice, including a video demonstration of the process:

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